In Vitro and In Vivo Anti-parasitic Activity of Sambucus ebulus and Feijoa sellowiana Extracts Silver Nanoparticles on Toxoplasma gondii Tachyzoites.

BACKGROUND: Current chemical treatments for toxoplasmosis have side effects, researchers are looking for herbal remedies with minimal side effects and the best effectiveness. This study aimed to evaluate the anti-toxoplasmic effects of silver nanoparticles based on Sambucus ebulus (Ag-NPs-S. ebulus) and Feijoa sellowiana (Ag-NPs-F. sellowiana) fruit extracts, in vitro and in vivo. METHODS: Vero cells were treated with different concentrations (0.5, 1, 2, 5, 10, 20, 40 µg/mL) of extracts and pyrimethamine as a positive control. Vero cells were infected with T. gondii and treated with extracts. The infection index and intracellular proliferation of T. gondii were evaluated. The survival rate of infected mice with tachyzoites of T. gondii was examined after intraperitoneal injection of the extracts at a dose of 40 mg/kg/day for 5 days after infection. RESULTS: The Ag-NPs-S. ebulus and Ag-NPs-F. sellowiana, almost similar to pyrimethamine, reduced proliferation index when compared to untreated group. Also, high toxoplasmicidal activity was observed with Ag-NPs-S. ebulus extract. Mice in the treatment groups of Ag-NPs-S. ebulus and pyrimethamine achieved better results in terms of survival than the others. CONCLUSION: The results indicated that Ag-NPs-F. sellowiana and S. ebulus have a significant growth effect on T. gondii in vitro and in vivo. Ag-NPs-S. ebulus extract has a more lethal effect on the parasite than Ag-NPs-F. sellowiana. It is suggested that in future investigate the induction of Toxoplasma-infected cell apoptosis using nanoparticles.

Enhancement of immune responses by vaccine potential of three antigens, including ROP18, MIC4, and SAG1 against acute toxoplasmosis in mice.

Toxoplasma gondii (T. gondii) causes considerable financial losses in the livestock industry and can present serious threats to pregnant women, as well as immunocompromised patients. Therefore, it is required to design and produce an efficient vaccine for controlling toxoplasmosis. The present study aimed to evaluate the protective immunity induced by RMS protein (ROP18, MIC4, and SAG1) with Freund adjuvant, calcium phosphate nanoparticles (CaPNs), and chitosan nanoparticles (CNs) in BALB/c mice. The RMS protein was expressed in Escherichia coli (E. coli) and purified using a HisTrap HP column. Thereafter, cellular and humoral immunity was assessed by injecting RMS protein on days 0, 21, and 35 into four groups [RMS, RMS-chitosan nanoparticles (RMS-CNs), RMS-calcium phosphate nanoparticles (RMS-CaPNs), and RMS-Freund]. Phosphate buffered saline (PBS), CNs, CaPNs, and Freund served as the four control groups. The results displayed that vaccination with RMS protein and adjuvants significantly elicited the levels of specific IgG antibodies and cytokines against toxoplasmosis. There were high levels of total IgG, IgG2a, and IFN-γ in vaccinated mice, compared to those in the control groups, especially in the RMS-Freund, indicating a Th-1 type response. The vaccinated and control mice were challenged intraperitoneally with 1 × 103 tachyzoites of the T. gondii RH strain four weeks after the last injection, and in RMS-Freund and RMS-CaPNs groups, the highest increase in survival time was observed (15 days). The RMS can significantly increase Th1 and Th2 responses; moreover, multi-epitope vaccines with adjuvants can be a promising strategy for the production of a vaccine against toxoplasmosis.

Anti-Toxoplasma Activities of the Hydroalcoholic Extract of Some Brassicaceae Species.

BACKGROUND: Toxoplasma gondii (T. gondii) is a protozoan parasite that infects a wide range of warm-blooded animals and humans. The conventional anti-Toxoplasma treatments cause significant toxicity. Brassicaceae family contains several medicinal plants with anti-inflammatory, chemopreventive, insecticide, antibacterial, antiviral, and antiparasitic effects. In this study, the hydroalcoholic extract of some Brassicaceae species was investigated against T. gondii in vitro. MATERIALS AND METHODS: Seeds of Alyssum homolocarpum, Lepidium perfoliatum, Lepidium sativum, and aerial parts of Nasturtium officinale and Capsella bursa-pastoris were extracted by maceration method using 80% ethanol. Vero cells were treated with different concentrations (5-600 µg/mL) of the extracts and pyrimethamine (as positive control), and the cellular viability was verified. Next, Vero cells were infected by T. gondii tachyzoites (RH strain), and the viability of the infected cells was measured by a colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. RESULTS: The 50% inhibitory concentration values were 5.1, 14.67, 32.49, 37.31, 71.35, and 2.63 µg/mL, and the selectivity indices were 8.06, 2.59, 0.74, 0.78, 0.65 (P < 0.05 compared with positive control), and 3.03 for L. sativum, L. perfoliatum, N. officinale, A. homolocarpum, C. bursa-pastoris, and pyrimethamine, respectively. CONCLUSION: The results of this study demonstrated that the hydroalcoholic extracts of L. sativum and L. perfoliatum have the promising anti-Toxoplasma activity by growth inhibition of T. gondii tachyzoites in infected cells.

Protective efficacy induced by DNA prime and recombinant protein boost vaccination with Toxoplasma gondii GRA14 in mice.

Toxoplasma gondii, the etiological agent of toxoplasmosis, can cause severe or lethal damages in both animals and man. So, tends to develop a more effective vaccine to prevent this disease is extremely needed and would be so prominent. The novel dense granule antigen 14 (GRA14) has been identified as a potential vaccine candidate against T. gondii infection. The aim of this study was evaluation of protective immunity induced by prime/boost vaccination strategy of GRA14 antigen with calcium phosphate (CaPNs) or Aluminum hydroxide (Alum) nano-adjuvants in BALB/c mice. The finding showed that immunization with the prime-boost strategy using plasmid DNA (pcGRA14) and recombinant protein (rGRA14) with nano-adjuvants significantly elicited levels of specific IgG antibodies and cytokines against T. gondii infection. Given that, there were the high levels of total IgG, IgG2a, IFN-γ in mice of rGRA14-CaPNs and pcGRA14 + rGRA14-CaPNs groups, which indicating a Th-1 type response. While immunization of mice with Alum based rGRA14 and pcGRA14 + rGRA14 elicited specific IgG1 and IL-4 levels, which was confirmed a Th-2 type response. Mice immunized with DNA prime-protein boost vaccine with nano-adjuvants produce more vigorous specific lymphoproliferative responses than mice immunized with other antigen formulations. In addition, the CaPNs-based prime-boost vaccine of pcGRA14 + rGRA14 showed the longest survival time in mice and the lowest parasitic load in their brain tissue compared to the other groups. The results obtained in this study show that the use of GRA14 based DNA prime-protein boost vaccination regime with CaPNs can dramatically enhanced both humoral and cellular immune responses. Therefore, this strategy can provide a promising approach to the development of an effective vaccine against T. gondii infection in the future.

Effects of Aloe vera and Eucalyptus methanolic extracts on experimental toxoplasmosis in vitro and in vivo.

Toxoplasmosis is a worldwide disease caused by the protozoan parasite Toxoplasma gondii (T. gondii), which is most commonly treated by pyrimethamine and sulfadiazine. However, this treatment presents several adverse side effects; Thus, new drugs with lower toxicities are urgently needed. In this study the anti-T. gondii activity of A. vera and Eucalyptus extracts were evaluated in vitro using a MTT (3-(4, 5-dimethylthiazol-2-yl) 2, 5-diphenyltetrazolium bromide) assay and in vivo by measuring the survival rates of mice infected with 2 × 103 tachyzoites of RH strain of T. gondii and then injected intraperitoneally by different concentrations of extracts for 4 days. Biochemical parameters such as Ferric Reducing Antioxidant Potential (FRAP) and malondialdehyde (MDA) assay were also evaluated. As results, in the in vitro assay, the IC50 values were 13.2, 24.7, 2.63 µg/ml, and the selectivity indexes were 3.3, 2.4, 3.03 for the A. vera, Eucalyptus and pyrimethamine, respectively. The mice treated with Eucalyptus showed a better survival rate than others (P < 0.05). The increased weight of liver and spleen, due to infection, was reduced by treatments. In FRAP assay Eucalyptus showed a better antioxidant activity than the other extracts. MDA levels in both liver and spleen were reduced by treatment. The results show that A. Vera and Eucalyptus possess anti-T. gondii activities in vitro and in vivo, in addition, Eucalyptus shows antioxidant activity with a higher survival rate. Therefore, Eucalyptus may be a useful candidate for treating Toxoplasma infection. Moreover, further studies are required to investigate the fractionations of this plant against T. gondii.

Natural products applied against hydatid cyst protoscolices: A review of past to present.

Echinococcus granulosus is the causative agent of cystic echinococcosis (CE), which is distributed all around the world. CE is one of the most important global parasitic infectious diseases, both in humans and animals. This parasite causes hydatid cysts that can be lodge at different organs of host such as liver, lung even in heart and brain which may lead to death. Presently, numerous scolicidal chemical agents have been administrated for inactivation of the hydatid cyst contents. Because of increasing resistance and adverse effects of medications include abnormalities of liver function, abdominal pain, diarrhea, nausea, vomiting, dizziness, and headache; there is a need to find alternative therapies either with the least or without side effects. Recently, there is a high tendency among researchers to evaluate and present herbal plants as alternative option due to being inexpensive, easy available, low side effects and toxicity. Till now, many efforts have been conducted on herbal extracts against protoscolices of hydatid cysts throughout the world. Therefore, the current review systematically searched the following electronic databases: PubMed, Science Direct, Scopus, and Google Scholar on published papers according to the keywords. In addition, a comprehensive list of medicinal plants was prepared and some of these herbal plants which showed the best efficacy and promising results are discussed elaborately.

Enhancing immune responses to a DNA vaccine encoding Toxoplasma gondii GRA14 by calcium phosphate nanoparticles as an adjuvant.

Several approaches have been used to improve the immunogenicity of DNA vaccines. In the current study, we constructed the plasmid encoding T. gondii dense granule 14 (GRA14) and investigated the immunological properties of calcium phosphate nanoparticles (CaPNs) as nano-adjuvant to enhance the protective efficacy of pcGRA14. BALB/c mice intramuscularly injected three times at two-week intervals and the immune responses were evaluated using lymphocyte proliferation assay, cytokine and antibody measurements, survival times, and parasite load of mice challenged with the virulent T. gondii RH strain. The results showed that the immune responses were induced in mice receiving pcGRA14 DNA vaccine. Interestingly, pcGRA14 coated with nanoparticles led to statistically significant enhancements of cellular and humoral immune responses against Toxoplasma infection (P<0.05). After challenge with RH strain of T. gondii, immunized mice with pcGRA14 showed prolong survival time compared to control groups (P<0.05). In addition, pcGRA14 coated with nano-adjuvant exhibited the lowest parasitic load in the infected mice tissues. For the first time, our data indicate that the pcGRA14 coated with CaPN was more effective for stimulation of immune responses and should be considered as an adjuvant in the design of vaccines against toxoplasmosis.

Immunological evaluation of a DNA cocktail vaccine with co-delivery of calcium phosphate nanoparticles (CaPNs) against the Toxoplasma gondii RH strain in BALB/c mice.

Many recent studies have been conducted to evaluate protective immunity mediated by DNA vaccines against toxoplasmosis. Cocktail DNA vaccines showed better immune responses compared to single vaccines. The objective of the current study was to evaluate the protective efficacy of rhomboid 4 (ROM4) and cocktail DNA vaccines (ROM4 + GRA14) of the Toxoplasma gondii RH strain with or without coated calcium phosphate nanoparticles (CaPNs) as the adjuvant to improve the immunogenicity against the T. gondii RH strain in BALB/c mice. Cocktail DNA vaccines of pcROM4 + pcGRA14 of the T. gondii RH strain were constructed. CaPNs were synthesized and the cocktail DNA vaccine was coated with the adjuvant of CaPNs. Immunogenicity and the protective effects of cocktail DNA vaccines with or without CaPNs against lethal challenge were evaluated in BALB/c mice. pcROM4 and cocktail DNA vaccine coated with CaPNs significantly enhanced cellular and humoral immune responses against Toxoplasma compared to pcROM4 and cocktail DNA vaccine without CaPNs (p < 0.05). These findings indicate that the survival time of immunized mice after challenge with the RH strain of T. gondii was increased compared to that of controls and the DNA vaccine provided significant protection in mice (p < 0.05). The CaPN-based cocktail DNA vaccine of pcROM4 + pcGRA14 showed the longest survival time compared to the other groups. Co-immunization with CaPN-based cocktail DNA vaccine (pcROM4 + pcGRA14) boosted immune responses and increased the protective efficacy against acute toxoplasmosis in BALB/c mice compared to both single gene and bivalent DNA vaccine without nano-adjuvants.

The efficacy of herbal medicines against Toxoplasma gondii during the last 3 decades: a systematic review.

The objective of the current study was to systematically review papers discussing the efficacy of medicinal herbs against Toxoplasma gondii. Data were systematically collected from published papers about the efficacy of herbs used against T. gondii globally from 1988 to 2015, from PubMed, Google Scholar, ISI Web of Science, EBSCO, Science Direct, and Scopus. Forty-nine papers were included in the current systematic review reporting the evaluation of medicinal plants against T. gondii globally, both in vitro and in vivo. Sixty-one plants were evaluated. Most of the studies were carried out on Artemisia annua. The second highest number of studies were carried out on Glycyrrhiza glabra extracts. RH and ME49 were the predominant parasite strains used. Additionally, Swiss-Webster and BALB/c mice were the major animal models used. Alcoholic and aqueous extracts were used more than other types of extracts. Natural compounds mentioned here may be developed as novel and more effective therapeutic agents that improve the treatment of toxoplasmosis due to their lower side effects, higher availability, and better cultural acceptance compared with those of the chemical drugs that are currently being used.

Anti-Toxoplasma activities of methanolic extract of Sambucus nigra (Caprifoliaceae) fruits and leaves / Actividad anti-Toxoplasma de extractos metanólicos de frutos y hojas de Sambucus nigra (Caprifoliaceae)

Toxoplasma gondii is an obligatory intracellular parasite that infects a wide range of warm-blooded animals and humans. Considering the severity of toxoplasmosis, side effects of current treatments, and the contribution of the ethnopharmacological knowledge for the treatment of parasitic infections, the aim of the present study was to investigate the efficacy of methanolic extracts from the fruits and leaves of Sambucus nigra against tachyzoite of T. gondii. For this, fruits and leaves of S. nigra were collected from Mazandaran province, Iran, were dried under the shade, and powdered using a commercial electrical blender. For extractions, methanol was used as solvent. Virulent RH strain of T. gondii was maintained in mice and macrophages containing tachyzoites were aspirated from the peritoneal cavity. Four concentrations (5, 10, 25 and 50mg/mL) of S. nigra extract were incubated with infected macrophages for 30, 60, 120 and 180 minutes and the viability of the tachyzoites were evaluated by trypan blue staining. Results showed that S. nigra fruit extracts at the concentrations of 5 and 10mg/mL killed 100% of T. gondii tachyzoites after 60 and 120 minutes, respectively; and concentrations of 25 and 50mg/mL killed 100% of the tachyzoites after 30 minutes. Additionally, extract of S. nigra leaves, at the concentrations of 5, 10 and 25mg/mL after 180 minutes, and concentration of 50mg/mL after 60 minutes, resulted with the highest efficacy. Our results showed that S. nigra has acceptable efficacy in vitro and the parasiticidal effect of fruit extract was significantly better than leaf extract. However, in vivo efficacy of this extract needs further investigation. Rev. Biol. Trop. 63 (1): 7-12. Epub 2015 March 01.

Toxoplasma gondii es un parásito intracelular obligatorio que infecta a una gran variedad de animales y seres humanos. Teniendo en cuenta la gravedad de la toxoplasmosis, los efectos secundarios de los tratamientos actuales, y la contribución de los conocimientos etnofarmacológicos para el tratamiento de infecciones parasitarias, el objetivo del presente estudio fue investigar la eficacia de los extractos metanólicos de los frutos y hojas de Sambucus nigra contra el taquizoito de T. gondii. Para esto, se recogieron frutos y hojas de S. nigra en la provincia de Mazandaran, Irán, se secaron a la sombra, y se pulverizaron con una batidora eléctrica comercial. Para las extracciones, se empleó metanol como disolvente. La cepa virulenta RH de T. gondii se mantuvo en ratones y los macrófagos con taquizoitos se aspiraron de la cavidad peritoneal. Cuatro concentraciones (5, 10, 25 y 50mg/ mL) de extracto de S. nigra se encubaron con los macrófagos infectados durante 30, 60, 120 y 180 minutos y la viabilidad de los taquizoitos se evaluó mediante tinción con azul de tripano. Los resultados mostraron que los extractos de frutos de S. nigra en las concentraciones de 5 y 10mg/mL mataron al 100% de los taquizoitos de T. gondii después de 60 y 120 minutos, respectivamente; y las concentraciones de 25 y 50mg/mL mataron al 100% de los taquizoitos después de 30 minutos. Además, el extracto de hojas de S. nigra, en concentraciones de 5, 10 y 25mg/mL después de 180 minutos, y una concentración de 50mg/mL después de 60 minutos, resultaron más eficientes. Nuestros resultados mostraron que S. nigra tiene una eficacia aceptable in vitro y el efecto parasiticida del extracto de frutos fue significativamente mejor que el del extracto de hoja. Sin embargo, la eficacia in vivo de este extracto necesita más investigación.

Homeostasis status between prooxidants and antioxidants as a potent marker in Iranian preeclamptic patients.

OBJECTIVE: To measure the plasma oxidant, lipid peroxidation and antioxidants, ascorbate, to magnify the prooxidants and antioxidants status as a marker of pre-eclampsia. METHODS: Included in the study were 50 preeclamptic and 100 normotensive pregnant women of singleton gestations in their third trimester, presented in Allavi Hospitals in Ardabil province of Iran from August 2004 to April 2005. Among these patients plasma malonaldehyde (MDA) and vitamin C concentration were analyzed. Blood samples (5 ml) were collected aseptically in heparin bulb. Spectrophotometric methods were employed to determine the plasma concentrations of vitamin C. The Statistical Package for Social Sciences software was used to analyze the data. RESULTS: In preeclampsia, significant increase in prooxidant MDA was observed as compared to controls. A significant fall in antioxidants vitamin C was noted in preeclampsia as compared to control. The MDA showed a significant balance to the level of vitamin C. CONCLUSION: Administration of regular nutritive supplementation containing reducing systems like vitamin C should help in maintaining the equilibrium during normal pregnancy and theoretically in preeclampsia too but to what extent they help in clinical practice is still being investigated.