Global prevalence of Toxoplasma gondii in birds: A systematic review and meta-analysis.

Among the potential animal reservoirs of the zoonotic parasite T. gondii, birds have received relatively little attention. This systematic review and meta-analysis aimed to assess the global status and to provide an overview of the epidemiology of T. gondii infection in birds. The standard protocol of preferred reporting items for systematic reviews and meta-analyses (PRISMA) guidelines were followed. Scopus, PubMed, Web of Science, Science Direct, ProQuest, and Google Scholar were searched for relevant publications from January 1990, to March 2024. All peer-reviewed original research articles describing the prevalence of T. gondii in birds were included. Inclusion and exclusion criteria were applied, and both direct and indirect detection were considered. The point estimates and 95% confidence intervals were calculated using the meta-package in R (version 3.6.1). The variance between studies (heterogeneity) was quantified by the I2 index. Finally, 258 articles (including 380 datasets) were eligible for inclusion in the systematic review and meta-analysis. The global pooled prevalence was 24% (21 - 26%). The highest prevalence of T. gondii was observed in buzzards (52%, 34 - 70%), turkeys (31%, 17 - 46%), and chickens (30%, 26 - 34%). The present study provides a comprehensive view of the global prevalence of T. gondii in birds.

Toxoplasma gondii infection and testosterone alteration: A systematic review and meta-analyses.

BACKGROUND: Toxoplasma gondii (T. gondii) is a worldwide distributed protozoan parasite which has infected a wide range of warm-blooded animals and humans. The most common form of T. gondii infection is asymptomatic (latent); nevertheless, latent toxoplasmosis can induce various alterations of sex hormones, especially testosterone, in infected humans and animals. On the other hand, testosterone is involved in behavioral traits and reproductive functions in both sexes. Hence, the purpose of this systematic review is to summarize the available evidence regarding the association between T. gondii infection and testosterone alteration. METHODS: In the setting of a systematic review, an electronic search (any date to 10 January 2023) without language restrictions was performed using Science Direct, Web of Science, PubMed, Scopus, and Google Scholar. The PRISMA guidelines were followed. Following the initial search, a total of 12,306 titles and abstracts were screened initially; 12,281 were excluded due to the lack of eligibility criteria or duplication. Finally, 24 articles met the included criteria. A mean±standard deviation (SD) was calculated to assess the difference of testosterone between T. gondii positive and T. gondii negative humans. The possibility of publication bias was assessed using Egger's regression. P-value < 0.05 was considered statistically significant. RESULTS: This systematic review identified 24 articles (18 studies in humans and six studies in animals). Most human studies (13 out of 19) reported an increased level of testosterone following latent toxoplasmosis in males, while three studies reported decreased levels and two studies reported an insignificant change. Eleven articles (seven datasets in males and seven datasets in females) were eligible to be included in the data synthesis. Based on the random-effects model, the pooled mean± SD of testosterone in T. gondii positive than T. gondii negative was increased by 0.73 and 0.55 units in males and females, respectively. The Egger's regression did not detect a statistically significant publication bias in males and females (p = value = 0.95 and 0.71), respectively. Three studies in male animals (rats, mice, and spotted hyenas) and two studies in female animals (mice and spotted hyenas) reported a decline in testosterone in infected compared with non-infected animals. While, one study in female rats reported no significant changes of testosterone in infected than non-infected animals. Moreover, two studies in male rats reported an increased level of testosterone in infected than non-infected animals. CONCLUSIONS: This study provides new insights about the association between T. gondii infection and testosterone alteration and identifies relevant data gaps that can inform and encourage further studies. The consequence of increased testosterone levels following T. gondii infection could partly be associated with increased sexual behavior and sexual transmission of the parasite. On the other hand, declining testosterone levels following T. gondii infection may be associated with male reproductive impairments, which were observed in T. gondii-infected humans and animals. Furthermore, these findings suggest the great need for more epidemiological and experimental investigations in depth to understand the relationship between T. gondii infection and testosterone alteration alongside with future consequences of testosterone alteration.

Antimalarial Effects of Nano Chloroquine Loaded Curcumin In vivo.

BACKGROUND: Malaria is still the deadliest parasitic disease caused by Plasmodium spp. Due to drug resistance and their unpleasant side effects, of conventional researchers are enormously seeking to achieve antimalarial drugs with more curative effective, less toxic and cost-affordable drugs using more advanced technology such as nanodrugs. PURPOSE: The present study aimed to examine the antimalarial effects of a novel synthesized nonochloroquine-loaded curcumin relying on dendrimer G2 in susceptible mice. METHODS: Antimalarial activity and toxicity of the nanocomposite were examined on BALB/C mice with microscopy, checking RBCs morphology and related enzymatic activity rate. RESULTS: The maximum inhibitory effect of the nanocomposite was seen at 10 mg/kg, killing 98% of P. berghei compared to sole chloroquine, whereas ED50 was reported at 5.5 mg/kg. The safety of the synthesized nanocomposite was confirmed with biochemical tests with no detrimental effects on mice. The sustainability and longevity of the nanodrug increased significantly with the NDC-CQ assay compared to the control groups. CONCLUSION: The study showed that nonochloroquine-loaded curcumin had a promising inhibitory effect on P. berghei growth in infected mice compared to standard drugs. However, further studies and clinical trials with large samples are recommended to study different aspects of using nanodrug.

In vitro and In vivo Effects of Ethanolic Extract of Fumaria parviflora Lam. Embedded in Chitosan Nanoparticles Against Leishmania major.

INTRODUCTION: Fumaria has been traditionally used to treat skin damages due to anti-inflammatory properties. In the present study, we evaluated the effect of the ethanolic extract of Fumaria parviflora Lam. (F. parviflora) against Leishmania major (L. major) using chitosan biopolymer drug delivery system both In vitro and In vivo models. MATERIALS AND METHODS: The ethanolic extract of F. parviflora was analyzed by HPLC to determine its active ingredients content. The extract was then loaded on chitosan nanoparticles (CNPs). The parasite was treated with various concentrations of the ethanolic extract, CNPs and CNPs loaded with F. parviflora extract (CNPs@ F. parviflora). The size of lesions of treated mice were measured on a weekly basis. The parasite burden was evaluated 8 weeks after treatment. RESULTS: The HPLC analysis showed the presence of Fumaric acid at a high concentration. The percentage of the drug released from CNPs@ F. parviflora within 24 and 72 h were 65% and 90% respectively. The results showed that F. parviflora extract and CNPs@ F. parviflora caused 84% and 96% growth inhibition of L. major promastigotes as revealed by Neubauer chamber counting and MTT test respectively. The IC50 values of F. parviflora extract and CNPs@ F. parviflora were 450 and 68.4 µg/ml respectively. In amastigote assay, the best results showed in CNPs@ F. parviflora that only 2% of macrophages were infected with amastigotes. In vivo experiments for mice treated with F. parviflora and CNPs @ F. parviflora in comparison to control group showed a significant reduction (P < 0.05) in the mean diameter of the lesions (2.3 and 1.72 mm and 9.91 mm respectively). CONCLUSION: The ethanolic extract of F. parviflora both as standalone and loaded in CNPs showed promising inhibitory effects against L. major both upon In vitro and In vivo experimentation as well as therapeutic effects for wound healing in infected mice.

Toxoplasma gondii infection and ABO blood groups: a systematic review and meta-analysis.

The current study aimed to determine the prevalence of Toxoplasma gondii in ABO blood groups and assess the relationship between the prevalence of T. gondii and blood groups. A literature search was carried out for epidemiological studies that were published through December 2022. A random effects model was used to determine the OR and the pooled prevalence with a 95% CI. The estimated pooled prevalences of T. gondii infection in the A, B, AB and O blood groups were 38% (95% CI 27 to 48%), 38% (95% CI 29 to 47%), 36% (95% CI 26 to 45%) and 36% (95% CI 27 to 45%), respectively. Also, the pooled ORs of the relationship between the prevalence of T. gondii infection and the A, B, AB and O blood groups were 1.08 (95% CI 0.97 to 1.19), 1.10 (95% CI 0.95 to 1.28), 1.08 (95% CI 0.92 to 1.27) and 0.89 (95% CI 0.80 to 1.00), respectively. This meta-analysis did not show any relationship between the prevalence of T. gondii infection and ABO blood groups.

In vitro Evaluation of the Potent Antileishmanial Activity of Ferula tabasensis Alone or in Combination with Shark Cartilage Extract Against the Standard Iranian Strain of Leishmania major (MRHO/IR/75/ER).

Background: The available drugs for the treatment of leishmaniasis are highly toxic and extremely expensive, with low efficiency; therefore, the development of effective therapeutic compounds is essential. Objectives: The present study aimed to explore the antileishmanial effects of ethyl acetate extract, methanol extract, and fractions 1-4 (F1-F4) of Ferula tabasensis, alone or in combination with shark cartilage extract (ShCE), on L. major in vitro. Methods: In this study, ethyl acetate, methanol, and n-hexane extracts were extracted from the aerial roots of F. tabasensis by the maceration method. The silica gel column chromatography was used to separate n-hexane extracts at varying polarities (F1-F4 fractions). Subsequently, the effects of extracts and fractions against promastigotes were assessed by the parasite counting method microscopic inhibition test and MTT assay. Besides, their effects on the infected macrophage cells and the number of amastigotes were investigated. Cytotoxicity was evaluated in non-infected J774A.1 macrophage cells. Finally, apoptosis induction of promastigotes, including infected and non-infected macrophages, was evaluated. Results: The results indicated the highly potent activity of F. tabasensis extracts and F1-F4 fractions, alone or in combination with ShCE, against L. major promastigotes and amastigotes in a dose-dependent manner (P < 0.05). The F1 fraction and methanol extract showed markedly higher toxicity compared to the other extracts and fractions, with 50% inhibitory concentrations (IC50/72h) of 2.4 ± 0.29 and 2.9 ± 0.55 µg/mL against promastigotes and 1.79 ± 0.27 µg/mL and 1.39 ± 0.27 µg/mL against amastigotes (P < 0.001). Moreover, they had a high selectivity index (SI) due to the low toxicity of macrophages (P < 0.0001). The results of flow cytometry indicated that the percentages of apoptotic promastigote cells in contact with IC50 concentrations of F1 and methanol extract alone after 72 h were 43.83 and 43.93%, as well as 78.4%, and 65.45% for their combination with ShCE, respectively.Also, apoptosis of infected macrophages induced by F1 and methanol extracts was estimated at 68.5% and 83.7%, respectively. Conclusions: In this study, the F1 fraction and methanol extract of F. tabasensis showed potent efficacy against L. major, associated with low toxicity and apoptosis induction. Therefore, they can be promising therapeutic candidates in future animal and even human studies.

Immune system roles in pathogenesis, prognosis, control, and treatment of Toxoplasma gondii infection.

Toxoplasma gondii is the protozoan causative agent of toxoplasmosis in humans and warm-blooded animals. Recent studies have illustrated that the immune system plays a pivotal role in the pathogenesis of toxoplasmosis by triggering immune cytokines like IL-12, TNF-α, and IFN-γ and immune cells like DCs, Th1, and Th17. On the other hand, some immune components can serve as prognosis markers of toxoplasmosis. In healthy people, the disease is often asymptomatic, but immunocompromised people and newborns may suffer severe symptoms and complications. Therefore, the immune prognostic markers may provide tools to measure the disease progress and help patients to avoid further complications. Immunotherapies using monoclonal antibody, cytokines, immune cells, exosomes, novel vaccines, and anti-inflammatory molecules open new horizon for toxoplasmosis treatment. In this review article, we discussed the immunopathogenesis, prognosis, and immunotherapy of Toxoplasma gondii infection.

Assessment of tissue levels of miR-146a and proinflammatory cytokines in experimental cerebral toxoplasmosis following atovaquone and clindamycin treatment: An in vivo study.

BACKGROUND: Despite recent advances for treating cerebral toxoplasmosis (CT), monitoring the parasite burden and treatment response is still challenging. miRNAs are small non-coding RNAs with regulatory functions that can be used in diagnosis and treatment monitoring. We investigated the changes in miR-146a, BAG-1 gene, IL-6, and IL-10 tissue levels in the brain of BALB/c mice with chronic CT caused by the PRU strain of T. gondii following anti-parasitic and antibiotic treatment. METHOD: Fifty-three 6-to 8-week-old BALB/c mice were infected using intraperitoneal inoculation of cerebral cysts of T. gondii PRU strain and then divided into five groups as follows: group 1 included mice treated with 100 mg/kg/d Atovaquone (AT), group 2 included mice treated with 400 mg/kg/d clindamycin (CL), group 3 included mice treated with combination therapy (AT + CL), group 4 included infected untreated mice as a positive control (PC), and; group 5 included uninfected untreated mice as negative control (NC). After the completion of the treatment course, tissue level of mir-146a, miR-155, BAG-1 gene, IL-6, and IL-10 was investigated with real-time polymerase chain reaction. The IL-6/IL-10 ratio was calculated as an indicator of immune response. Moreover, brain cyst numbers were counted on autopsy samples. RESULTS: miR-146a, IL-6, IL-10, and BAG-1 genes were expressed in PC, but not in the NC group; miR-146a, IL-6, IL-10, and BAG-1 gene expression were significantly lower in AT, CL, and AT + CL compared with PC. MiR-146a and BAG-1 levels in AT and CL were not different statistically, however, they both had lower levels compared to AT + CL (P < 0.01). There was no difference in the expression of IL-6 and IL-10 between treatment groups. BAG-1 expression was significantly lower in AT, than in CL and AT + CL (P < 0.0089 and < 0.002, respectively). The PC group showed a higher ratio of IL-6/IL-10, although this increase was not statistically significant. It is noteworthy that the treatment with AT reduced this ratio; in the inter-group comparison, this ratio showed a decrease in the AT and AT + CL compared to the PC. The number of brain tissue cysts was significantly lower in AT, CL, and AT + CL, than in PC (p < 0.0001). AT had significantly lower brain cysts than CL and AT + CL (P < 0.0001). CONCLUSION: It seems that the factors studied in the current research (microRNA and cytokines) are a suitable index for evaluating the response to antiparasitic and antibiotic treatment. However, more studies should be conducted in the future to confirm our findings.

Berberine improves inhibitory avoidance memory impairment of Toxoplasma gondii-infected rat model of ketamine-induced schizophrenia.

BACKGROUND: Memory impairment caused by Toxoplasma gondii infection has been documented. Berberine (BRB) is well known for its enhancing effects on memory and has shown promising results. However, the impact of BRB on T. gondii infection and schizophrenia-induced consolidation and reconsolidation memory impairment is still unclear. Here; we examined the effect of BRB on the inhibitory avoidance (IA) memory consolidation and reconsolidation impairment induced by T. gondii infection, and ketamine (Ket) as a pharmacological model of schizophrenia. Also; the brain-derived neurotrophic factor (BDNF) levels in the medial prefrontal cortex (mPFC) and hippocampus were analyzed. METHODS: Rats were infected with T. gondii RH strain or received Ket (30 mg/kg/day) intraperitoneally (i.p) for at least five consecutive days (as the model of schizophrenia). Then followed by oral administration with BRB (25 mg/kg/day) for five days. Finally, the IA memory retention test was examined 48 post-conditioning, and BDNF was measured. RESULTS: Results indicated IA memory impairment in T. gondii-infected animals since lower step-through latency (STL) was observed than in control animals. We found significant (P = 0.01, P = 0.001) elevations in STL and a significant decrease (P = 0.001) in total time spent in the dark area following BRB administration in infected and Ket-treated rats, indicating improvement (increased STL) in consolidation and reconsolidation memory. Moreover, BDNF levels were reduced (P = 0.01) in the hippocampus and mPFC regions of both T. gondii- infected and Ket-induced groups, which remarkably enhanced after BRB treatment. Furthermore; we found that BRB administration notably increased the mPFC BDNF levels in mPFC (P < 0.01) and hippocampus (P = 0.001) in the Ket-treated and rats infected with T. gondii. CONCLUSION: Taken together; BRB may be a valuable preclinical treatment for improving memory impairment through BDNF expression in PFC and hippocampus, therefore; BRB is suggested for memory disturbances induced by T. gondii infection.

Evaluating leishmanicidal effects of Lucilia sericata products in combination with Apis mellifera honey using an in vitro model.

Leishmaniasis is a zoonotic disease caused by an intracellular parasite from the genus Leishmania. Lack of safe and effective drugs has increasingly promoted researches into new drugs of natural origin to cure the disease. The study, therefore, aimed to investigate the anti-leishmanial effects of Lucilia sericata larval excretion/secretion (ES) in combination with Apis mellifera honey as a synergist on Leishmania major using an in vitro model. Various concentrations of honey and larval ES fractions were tested against promastigotes and intracellular amastigotes of L. major using macrophage J774A.1 cell line. The inhibitory effects and cytotoxicity of ES plus honey were evaluated using direct counting method and MTT assay. To assess the effects of larval ES plus honey on the amastigote form, the rate of macrophage infection and the number of amastigotes per infected macrophage cell were estimated. The 50% inhibitory concentration (IC50) values were 21.66 µg/ml, 43.25 60 µg/ml, 52.58 µg/ml, and 70.38 µg/ml for crude ES plus honey, ES >10 kDa plus honey, ES <10 kDa plus honey, and honey alone, respectively. The IC50 for positive control (glucantime) was 27.03 µg/ml. There was a significant difference between viability percentages of promastigotes exposed to different doses of applied treatments compared to the negative control (p≤ 0.0001). Microscopic examination of amastigote forms revealed that dosages applied at 150 to 300 µg/ml significantly reduced the rate of macrophage infection and the number of amastigotes per infected macrophage cell. Different doses of larval products plus honey did not show a significant toxic effect agaist macrophage J774 cells. The larval ES fractions of L. sericata in combination with A. mellifera honey acted synergistically against L. major.

Toxoplasma infection and Rhesus blood group system: A systematic review and meta-analysis.

BACKGROUND: Toxoplasmosis is one of the most common infections in humans and animals, which is caused by an obligate intracellular opportunistic parasite known as Toxoplasma gondii (T. gondii). Some data have shown that both Rhesus (Rh)-positive and Rh-negative individuals differ in response to biological factors, including Toxoplasma infection. Therefore, this systematic review and meta-analysis was conducted to investigate the scientific evidence regarding the possible association between the Rh blood group and Toxoplasma infection and to determine the seroprevalence of T. gondii in the Rh blood group system. METHODS: The research was conducted on PubMed, ScienceDirect, ProQuest, and Google Scholar databases until January 2023. Twenty-one cross-sectional studies were included with a total of 10910 people. The data were synthesized using a random effect model with 95% confidence intervals (CIs). RESULTS: The overall prevalence of T. gondii was calculated at 32.34% (CI 95%: 28.23-36.45%) and 33.35% (CI 95%: 19.73-46.96%) in Rh-positive and Rh-negative blood groups. In addition, the pooled OR for the relationship between the Rh blood group and the seroprevalence of T. gondii was 0.96 (95% CI: 0.72-1.28). CONCLUSIONS: This meta-analysis showed a high prevalence of Toxoplasma infection in both Rh-negative and positive blood groups. This systematic review and meta-analysis revealed that no significant association was found between toxoplasmosis and Rh factor. Because of the limited number of studies in this field, more research is recommended to determine the exact relationship between toxoplasmosis and the Rh factor.

Thymoquinone Antifungal Activity against Candida glabrata Oral Isolates from Patients in Intensive Care Units-An In Vitro Study.

The number of Candida spp. infections and drug resistance are dramatically increasing worldwide, particularly among immunosuppressed patients, and it is urgent to find novel compounds with antifungal activity. In this work, the antifungal and antibiofilm activity of thymoquinone (TQ), a key bioactive constituent of black cumin seed Nigella sativa L., was evaluated against Candida glabrata, a WHO 'high-priority' pathogen. Then, its effect on the expression of C. glabrata EPA6 and EPA7 genes (related to biofilm adhesion and development, respectively) were analyzed. Swab samples were taken from the oral cavity of 90 hospitalized patients in ICU wards, transferred to sterile falcon tubes, and cultured on Sabouraud Dextrose Agar (SDA) and Chromagar Candida for presumptive identification. Next, a 21-plex PCR was carried out for the confirmation of species level. C. glabrata isolates underwent antifungal drug susceptibility testing against fluconazole (FLZ), itraconazole (ITZ), amphotericin B (AMB), and TQ according to the CLSI microdilution method (M27, A3/S4). Biofilm formation was measured by an MTT assay. EPA6 and EPA7 gene expression was assessed by real-time PCR. From the 90 swab samples, 40 isolates were identified as C. glabrata with the 21-plex PCR. Most isolates were resistant to FLZ (n = 29, 72.5%), whereas 12.5% and 5% were ITZ and AMB resistant, respectively. The minimum inhibitory concentration (MIC50) of TQ against C. glabrata was 50 µg/mL. Importantly, TQ significantly inhibited the biofilm formation of C. glabrata isolates, and EPA6 gene expression was reduced significantly at MIC50 concentration of TQ. TQ seems to have some antifungal, antibiofilm (adhesion) effect on C. glabrata isolates, showing that this plant secondary metabolite is a promising agent to overcome Candida infections, especially oral candidiasis.

Investigation of antileishmanial activities of CaO nanoparticles on L. tropica and L. infantum parasites, in vitro.

Stibogluconate sodium and meglumine antimoniate are the main antimonials utilised as the primary treatment option for leishmaniasis. However, have a number of side effects that limit their use. Development of nanoparticles (NPs) use in biological research and remarkable antimicrobial effects and unique optical and structural properties of CaO NPs have motivated this study to evaluated the effect of different times/dilutions of CaO NPs on Leishmania tropica and Leishmania infantum. To evaluate the antileishmanial activity of CaO NPs, the cytotoxic effect of CaO NPs against L. tropica and L. infantum amastigotes, promastigotes, as well as macrophages, was evaluated using counting and MTT assay after adding different concentrations of CaO nanoparticles (800-6.25 µg/ml) to the parasite culture. The possible apoptosis by CaO NPs were evaluated via flow cytometry assay. The XRD-pattern related to CaO nanoparticles indicating the cubic phase structures. According the effects of nanoparticle on promastigotes the IC50 values of CaO nanoparticles within 72 h were 19.81 µg/ml for L. tropica and 22.57 µg/ml for L. infantum. The percentage of the normal, apoptotic, and necrotic cells was estimated to be 82.6%, 14.81%, and 2.69% for L. tropica, and 73.6%, 23.89%, and 2.58% for L. infantum, respectively. Our results showed acceptable in vitro activity level of CaO NPs against L. tropica and L. infantum promastigotes as well as intracellular amastigotes. CaO NPs were more effective against L. infantum compared to L. tropica in vitro study.

Molecular detection and characterisation of Theileria in hard ticks of small ruminants in Zarrin Dasht County, Southern Iran.

BACKGROUND: Ticks are important ectoparasites of small ruminants in tropics and subtropics including Iran. They transmit serious zoonotic pathogens such as Babesia and Theileria. These parasites cause major burden on small ruminants jeopardising livelihoods of rural people in Zarrin Dasht County. OBJECTIVES: This study was carried out to investigate the diversity and distribution of hard ticks of small ruminants and their piroplasm infection in a bid to contribute to Theileria and/or Babesia detection and control in Zarrin Dasht County of Fars province, Iran. METHODS: We examined 751 sheep and goats from 10 sites of the County during four seasons for hard tick infestation. The collected hard ticks (994) were taxonomically identified before being separately confined in microtubes coded to indicate their species and host animals as well as site and date of collection. In total 50 pooled samples were analysed by PCR technique for Theileria and Babesia infection. RESULTS: The identified ticks included Hyalomma marginatum 994/362); 36.4%), Rhipicephalus turanicus 994/352); 35.51%), Hyalomma anatolicum 994/264); 26.6%), Hyalomma dromedarii 994/14); 1.41%) and Hyalomma asiaticum 994/2) 0.2%). Molecular analyses showed that 7 out of 50 pooled sample were infected with piroplasm genome in ticks shared by Theileria ovis (6:50) and Theileria lestoquardi (1:50). Babesia was absent in collected hard ticks. CONCLUSIONS: This is the first report on the presence of piroplasm infection in hard ticks of small ruminants in Zarrin Dasht County. Theileria ovis was more prevalent than Theileria lestoquardi but Babesia was absent. Piroplasm infection was detected in Hyalomma marginatum, Hyalomma anatolicum and Rhipicephalus turanicus. Hyalomma marginatum appears to be more competent to vector Theileria spp. This study may contribute to risk assessment and prevention of epizootic theileriosis in the County.

In vitro and in vivo study on antiprotozoal activity of calcium oxide (CaO) and magnesium oxide (MgO) nanoparticles on promastigote and amastigote forms of Leishmania major.

BACKGROUND: Currently, anti-leishmanial drugs have been developed. However, the available compounds have several side effects such as drug resistance and toxicity that cause some limitation for use. The development of nanoparticles (NPs) use in biological research and the proven effectiveness of CaONPs and MgONPs on bacteria and fungi, along with the lack of information about its antileishmanial effects, have motivated this study. CaO and MgONPs possess considerable antibacterial effects because of their alkalinity and active oxygen species. This study has taken into account the impacts of these two NPs on the L. major in vitro and in vivo. METHODS: To evaluate the antileishmanial activity of NPs, the cytotoxic effect of CaONPs, MgONPs, and MgOCaONPs against L. major amastigotes, promastigotes, as well as macrophages, was evaluated using counting or MTT assay. The possible apoptosis of L. major by CaONPs, MgONPs, and MgOCaONPs was evaluated via flow cytometry assay. For in vivo study, BALB/c mice were allocated to five groups and the lesions of infected mice with L. major promastigotes were treated with a 200 µg/mL concentration CaONPs, MgONPs, and MgOCaONPs, then the mice underwent a 4-week follow-up to examine the wound diameter and survival rates. RESULTS: The XRD-pattern related to CaONPs and MgONPs indicating the cubic phase and Rocksalt cubic structures. According the effects of nanoparticle on promastigotes the IC50 values of CaONPs, MgONPs, and MgOCaONPs within 72 h were 7.9 ug/mL, 10.3 ug/mL, and 8.0 ug/mL respectively. CaONPs, MgONPs, and MgOCaONPs induced apoptosis in about 7.8%, 53.57%, and 12.8% of promastigotes. All mice presented lesions. MgONPs was the most effective in reducing the size of the lesions. CONCLUSION: According to the results of the present research, MgONPs and CaONPs showed good in vitro and in vivo effects on L. major promastigotes and intracellular amastigotes especially MgONPs, and also it seems that MgONPs are applicable in Leishmania infection treatment due to their potential antileishmanial effects.

In Vitro Activity of Green Synthesized Silver Nanoparticles via Thymus Vulgaris Extract against Leishmania major.

Background: We aimed to assess the in vitro effects of the green synthesized silver nanoparticles (Ag NPs) via Thymus vulgaris (thyme) against Leishmania major infection. Methods: We have prepared T. vulgalis silver nanoparticles (TSNPs) by adding thyme extract to the silver nitrate aqueous solution (0.2 mM), and evaluated their antileishmanial activity. The viability of L. major promastigotes was assessed in the presence of various concentrations of TSNPs by direct counting after 24 h. The MTT assay was used to identify the viability of promastigotes. The same procedures were assessed in uninfected macrophage cells. The apoptotic effects of nanoparticles on L. major promastigotes were determined by flow cytometry assay using annexin staining. To evaluate anti-amastigotes activity of TSNPs, light microscopic observation was used to determine the number of parasites within the macrophages in each well. Results: The effect of TSNPs on promastigotes and amastigotes of L. major was effective and had a reverse relationship with its concentration. TSNPs, inhibited the growth rate of L. major amastigotes and, the IC50 value of these nanoparticles was estimated 3.02 µg/mL (28 µM) after 72h. The results of flow cytometry showed that the toxic effects of TSNPs on promastigotes after 24 hours were statistically significant (P<0.05) and showed 69.51% of apoptosis. Conclusion: TSNPs had an inhibitor effect on promastigote and amastigote forms of L. major in vitro. It might be considered as a candidate for the treatment of this infection.

Protective immunity induced with a DNA vaccine encoding B- and T-cells multi-epitope SAG1, ROP16, MIC4, GRA12, M2AP, and multi-epitope ROP8 against acute and chronic toxoplasmosis in BALB/c mice.

BACKGROUND: T. gondii infection is characterized by a high global prevalence. Nearly, 16-40% of people have been infected by T. gondii. Although T. gondii often causes subclinical infection, it may cause severe complications in newborns with congenital infection and immunocompromised individuals. Constant attempts of scientists have made valuable findings in the development of T. gondii candidate vaccines. However, an effective vaccine has not been successfully developed yet. In this study, multi-epitope SAG1, MIC4, ROP16, M2AP, GRA12, and multi-epitope ROP8 were injected into BALB/c mice intramuscularly, as cocktailed plasmids or as single-gene plasmids to assess the immune response against chronic and acute Toxoplasma infection. METHODS: BALB/c mice were immunized on days 0, 21, and 42. The immune responses of both vaccinated and control groups were evaluated using cytokine and antibody measurements, lymphocyte proliferation assay, survival time, and average number of cysts in each brain. RESULTS: The results indicated that DNA vaccination using multi-epitope ROP8 and multi-epitope SAG1, ROP16, MIC4, GRA12, M2AP could elicit both cellular and humoral immune responses, and enhanced the survival time in BALB/c mice. Also, the administration of multi-epitope ROP8 plus multi-epitope SAG1, ROP16, MIC4, GRA12, M2AP could enhance the concentrations of IgG antibody, elicit a mixed IgG1/IgG2a reaction with the predominance of the IgG2a, increase the release of IFN-γ cytokine, prolonge the survival time, and reduce the brain cysts. CONCLUSIONS: Here, we report that vaccination using cocktailed plasmids could induce better protective immunity compared to single plasmid for acute and chronic T. gondii infection.

Treatment of Murine Toxoplasmosis with Oral and Parenteral Artemether and Following by Detection of B1 Gene by Quantitative Real-Time PCR (qPCR) for Evaluating Parasite Density.

Background: Toxoplasma gondii is an obligate intracellular parasite that can infect humans and animals. As the choice drug have shown side effects, development a new drug with low toxicity will be necessary. Methods: BALB/c mice were infected with tachyzoiets of T. gondii. After treatment by oral and parenteral artemether (250 µg/mice) and sulfadiazine (50 µg/mice), we evaluated the rates of survival in treated and control mice. The fold change of B1 gene (target gene) expression in liver and brain of mice treated with parenteral artemether (i.p.), oral artemether (via gavage) and sulfadiazine, were detected by using the Real-Time quantitative PCR. Results: Both treatment with sulfadiazine and artemether showed significant prolongation in time to death of the infected mice compared to the control group. Median survival days for parenteral artemether, oral artemether, sulfadiazine and control group were 8, 11, 12 and 6 d respectively. Expression of B1 gene in liver and brain of mice after treatment with artemether and sulfadiazine were reduced in comparison to housekeeping gene (ß-tubulin gene). The fold change (comparing to control group) for parenteral artemether, oral artemether, sulfadiazine is 0.034, 0.027 and 0.111 for liver and 0.220, 0.425 and 0.366 for brain respectively. Conclusion: Artemether is effective to control the tachyzoites of T. godii in vivo conditions and oral treatment is more effective than parenteral treatment. Due to its low cytotoxicity and its high effective action against the tachyzoietes of T. godii in susceptible animals.

Anticancer Activity of Hydatid Cyst Fluid along with Antigen B on Tumors Induced by 4T1 Breast Cancer Cell in a BALB/c Mice Model.

Background: Studies on experimental model of cancer showed that hydatid cyst fluid (HCF) has antitumor activity. We aimed to investigate the effect of HCF and Antigen B (AgB) on 4T1 breast tumor cells in BALB/c mice. Methods: This study was carried out in the Department of Parasitology, Tarbiat Modares University, Tehran, Iran from 2019 to 2020. There were two control groups of BALB/c mice (one group were injected with aluminum sulfate and another group with PBS), and six groups, injected via the intraperitoneal route with 100, 300 and 500 µg/ml concentrations of HCF, AgB diluted in 100 µl PBS, and alum. Seven days after the last treatment, 7×105 4T1 cells were subcutaneously injected into the right flank of BALB/c mice. Results: The difference between the mean size of the tumor in the case and control groups was statistically significant (P<0.05). The mean level of cytokines between the case and control groups was statistically significant (P<0.05). Our histo-pathological studies showed a reduction in both tumor volume and carcinogenesis in groups injected with 300 µg/ml HCF and 500 µg/ml AgB. The antitumor activity of HCF and AgB may be related to the immune responses against these antigens. Conclusion: We suggest that polarization of the Th1/Th2 ratio toward the Th1 pathway occurred in groups injected with HCF and AgB. More comprehensive and precise experiments using different hydatid cyst components are required to investigate their prophylactic effects on breast cancer.

Spontaneous abortion among Toxoplasma gondii IgG seropositive women: Molecular detection, genotype identification, and serological assessment with conventional ELISA and avidity ELISA.

OBJECTIVES: It has been generally believed that women who exposed to Toxoplasma gondii before pregnancy and have anti-T. gondii IgG antibody are immunized and their newborns will be protected from congenital infection. This study is aimed to investigate the role of T. gondii infection in spontaneous abortion through serological and molecular methods in southern Iran. STUDY DESIGN: Blood samples were taken from 50 spontaneously aborted mothers and anti-T. gondii antibodies were assessed using conventional enzyme-linked immunosorbent assay (ELISA) and avidity ELISA methods. The placenta and blood samples of aborted women were used for detection of the parasite's DNA by polymerase chain reaction (PCR) method targeting the RE gene. The parasite genotypes were determined by PCR-restriction fragment length polymorphism (RFLP) method using SAG3 and GRA6 genes. RESULTS: IgG antibody was detected in 28% (14/50) of mothers, but all samples were negative for IgM antibody. In the avidity ELISA test, 26% (13/50) of the samples had a high avidity index, suggesting chronic infection, while a low avidity index was detected in one case (2%), which suggests acute infection. The parasite's DNA was detected in 18% (9/50) and 14% (7/50) of blood and placenta samples, respectively. All DNA positive samples were IgG positive. All isolates were belonged to the T. gondii type III genotype. CONCLUSION: The results suggest that T. gondii seropositive women are not protected from congenital transmission. However, the results should be interpreted cautiously until further studies will be confirmed these results.