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.

A global systematic review and meta-analysis on the babesiosis in dogs with special reference to Babesia canis.

BACKGROUND: Canine babesiosis is a clinically significant tick-transmitted disease caused by several species of the intraerythrocytic protozoan parasite Babesia, which result in a wide range of clinical manifestations, from mild, transient infection to serious disease and even death. OBJECTIVES: The current study aimed to estimate the global prevalence and associated risk factors of Babesia in dogs. METHODS: Multiple databases (PubMed, Scopus, ProQuest, Web of Science and Google Scholar) were searched for relevant literature published from January 2000 up to December 2022. The statistical analyses were performed based on the R software (version 3.6) meta-package. RESULTS: Out of 23,864 publications, 229 studies met the inclusion criteria. The pooled prevalence of canine babesiosis was 0.120 (95% CI; 0.097-0.146). The highest pooled prevalence was found in Europe (0.207, 95% CI; 0.097-0.344). Among several species, Babesia canis was the most prevalent parasite (0.216, 95% CI; 0.056-0.441). The highest pooled prevalence of Babesia in dogs was observed in the summer season (0.097, 95% CI; 0.040-0.174). CONCLUSIONS: Regular screening and appropriate control strategies are recommended for the prevention of transmission of tick-borne disease transmission among dogs.

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.

The global prevalence of Spirometra parasites in snakes, frogs, dogs, and cats: A systematic review and meta-analysis.

BACKGROUND: Spirometra infection is aneglected food- and waterborne disease with worldwide distribution. OBJECTIVES: The present study aims to estimate the global prevalence of Spirometra species in snakes, frogs, dogs and cats. METHODS: Multiple databases (PubMed, Scopus, ProQuest, Web of Science and Google Scholar) were searched for relevant literatures published up to March 2022. RESULTS: Among 131 data sets (including 113 articles) that met the inclusion, 15 investigations reported Spirometra infection in snakes, 23 in frogs, 41 in dogs and 52 in cats. The pooled prevalence (95% confidence interval) in intermediate hosts and definitive hosts was found to be 0.313% and 0.089%, respectively. Based on continent, the infection was most prevalent in Asia for studies on snakes (0.696%) and frogs (0.181%), while Africa (0.224%) and Oceania (0.203%) were the regions with the highest pooled prevalence rates of the infection in dogs and cats, respectively. Among different diagnostic methods, the highest pooled prevalence was related to morphological method for studies on snakes, frog and cats with rate of 0.665%, 0.189% and 0.104%, respectively. Regarding studies on dogs, the highest pooled prevalence was observed for molecular technique (0.101%). CONCLUSIONS: The results presented here revealed the importance of establishing a prevention and control measure focused on protection of aquaculture systems from being contaminated with faeces of dogs and cats, and raising awareness of parasitic zoonotic diseases to decrease the transmission risk.

Testosterone Augments Propagation of Toxoplasma gondii in Glioblastoma Cells In Vitro.

PURPOSE: Toxoplasmosis can induce various hormonal and behavioral alterations in humans and rodents. Previous studies revealed alterations of sex hormones; especially testosterone, in infected humans and rodents, but little is known about the effects of sex hormones on the propagation of T. gondii. Hence, we aimed to investigate whether testosterone and progesterone influence on T. gondii propagation in neural cells. METHODS: The glioblastoma cells (U-87MG) were treated with different concentrations of testosterone and progesterone and the infection was done by tachyzoites of the RH strain of T. gondii. The number of infected cells, viability of T. gondii-infected cells, and parasite burden were measured by direct counting under a light microscope, MTT assay, and quantitative real-time PCR (qPCR), respectively. RESULTS: The results showed that testosterone at concentrations of 100 and 250 nM significantly increased the number of infected cells and parasite burden 24 and 48 h post-treatment compared to untreated controls. Progesterone had no significant effects in the same manner. CONCLUSION: The results indicated that testosterone could augment the propagation of T. gondii in in vitro.

Global molecular epidemiology of microsporidia in pigs and wild boars with emphasis on Enterocytozoon bieneusi: A systematic review and meta-analysis.

BACKGROUND: Microsporidia are spore-forming intracellular pathogens with worldwide prevalence, causing emerging infections in humans and animals. Enterocytozoon bieneusi is a zoonotic species of microsporidia and is responsible for more than 90% of cases of microsporidiosis in humans and animals. Pigs and wild boars are important animal reservoirs of microsporidia. Hence, we aimed to estimate the global prevalence of microsporidia and genetic diversity of E. bieneusi in pigs and wild boars through a set of systematic review and meta-analysis (PRISMA) guidelines. METHODS: Four databases (Web of Science, PubMed, Scopus and Google Scholar) were searched between January 1, 2000 and April 30, 2021. Regarding meta-analysis, the random-effect model was employed by forest plot with 95% confidence interval (CI). RESULTS: After exclusion of irrelevant articles and duplication removal, 33 papers, including 34 datasets (30 datasets for domestic pigs and 4 for wild boars) finally meet the inclusion criteria to undergo meta-analysis. The pooled prevalence rates of microsporidia infection in domestic pigs and wild boars were 37.6% (95% CI: 30.8-44.9%) and 8.1% (95% CI: 2.1-26.8%), respectively. While, the pooled prevalence rates of E. bieneusi were 35% (95% CI: 28.4-42.2%) in domestic pigs and 10.1% (95% CI: 1.7-42.4%) in wild boars. The genotypes EbpA was the most reported genotype in domestic pigs and wild boars. Male animals had higher prevalence rates of microsporidia infection than females (27 vs. 17.4%, OR = 1.91; 95% CI, 0.77-4.71%). CONCLUSION: This study indicates the important role of domestic pigs and wild boars as animal reservoir hosts of microsporidia. Thereby, strategies for control and prevention of these zoonotic pathogens should be designed in pigs and wild boars.

Efficacy of green synthesized silver nanoparticles via ginger rhizome extract against Leishmania major in vitro.

INTRODUCTION: Leishmaniasis is a major public health problem that causes by parasite of the genus Leishmania. The pentavalent antimonial compounds that used for treatment are not safe or effective enough. The aim of the present study was preparation and evaluation of the efficacy of green synthesized silver nanoparticles against Leishmania major (L. major) in vitro. METHODS: To synthesis silver (Ag) nanoparticles (NPs), ginger extract was added to the 0.2mM AgNO3 aqueous solution (1:20). Effects of different concentrations of Ag-NPs on the number of L. major promastigotes were investigated using counting assay. The MTT test was applied to determine the toxicity of Ag-NPs on promastigotes of L. major, as well as, macrophage cells. Then, to evaluate the anti-amastigotes effects of Ag-NPs, parasites within the macrophages were counted by light microscope. Furthermore, to determine the induced apoptosis and necrotic effects of Ag-NPs on promastigotes, flow cytometry method was employed using annexin staining. RESULTS: The effect of Ag-NPs on promastigotes and amastigotes of L. major was effective and has a reverse relationship with its concentration. According to the results of anti-amastigote assay, the IC50 value of this nanoparticle was estimated 2.35 ppm after 72h. Also, Ag-NPs caused Programmed Cell Death (PCD) in promastigotes of L. major and showed 60.18% of apoptosis. DISCUSSION: Based on the mentioned results, it can be concluded that Ag NPs has a beneficial effect on promastigote and amastigote forms of L. major in vitro. Hence, these nanoparticles could be applied as promising antileishmanial agents for treatment of Leishmania infections.

Effect of Imiquimod on Tachyzoites of Toxoplasma gondii and Infected Macrophages in vitro and in BALB/c Mice.

Treatment for toxoplasmosis is not completely successful because of their unwanted side effects, and new treatments are needed. Imiquimod has ability to moderate immune response and used to treat a wide variety of infections and tumors. The aim of the present study was to evaluate the effect of imiquimod on the tachyzoites of T. gondii and infected macrophages in vitro and in BALB/c mice. The viability of T. gondii was assessed in the presence of various concentrations of imiquimod by direct counting after 6 and 24 h. The MTT assay was used to identify the viability of uninfected macrophages. The apoptotic effects were determined with flow cytometry on the tachyzoites and infected macrophages. For evaluation of parasite load in pre-treatment or post-treatment of macrophages Quantitative real time PCR (qPCR) was performed. For in vivo experiments, BALB/c mice received imiquimod before and after challenge with parasites. The mortality rate of mice, parasite numbers in spleen, and the INF-γ and IL-4 cytokine levels in spleen lymphocytes were evaluated. Imiquimod demonstrated anti-Toxoplasma effects by reducing the number of tachyzoites. The results of flow cytometry for drug-treated tachyzoites showed that apoptosis did not rise significantly relative to the control group (p < 0.05). Moreover, apoptosis was enhanced in infected macrophages as the concentration of imiquimod was reduced. The parasitic burden in imiquimod pretreated macrophages was significantly lower than those treated after infection (p < 0.01). A marked reduction was observed in survival rate, parasite load and INF-γ level in BALB/c mice that received imiquimod before parasitic challenge relative to those received drug after parasitic challenge (p < 0.01). Overall, imiquimod in the pretreated group had greater anti-Toxoplasma effects than imiquimod in posttreated group in vitro and in vivo. imiquimod may be considered as a candidate for use against Toxoplasmosis both therapeutically and prophylactically.

Toxoplasma gondii: Preventive and therapeutic effects of morphine and evaluation of treatment parameters of tachyzoites and infected macrophages in vitro and in a murine model.

Common medicines for the treatment of toxoplasmosis have limited efficacy and unwanted side effects. Opiates can effect both innate and cell-mediated immunity and stimulate the immune responses in different parasitic infections. In this work, preventive and therapeutic effects of morphine were evaluated on the tachyzoites of Toxoplasma gondii and infected macrophages in vitro and in a murine model. Different concentrations of morphine (0.1 and 0.01 µg/ml) were evaluated on mortality rate of T. gondii by direct counting after 3 and 24 hours. The cytotoxic and apoptotic effects of these drugs were measured by the MTT assays and flow cytometry analysis, respectively. The same procedures were assessed in T. gondii-infected macrophages. The parasite loads were determined using quantitative polymerase chain reaction (qPCR). For in vivo assessment, BALB/c mice treated with morphine before or after infection with tachyzoites. The survival rate of animals, parasite load in the spleen, and the IFN-γ and IL-4 cytokines levels were measured. Morphine was effective on tachyzoites of T. gondii and had a reverse relationship with its concentration. The results of flow cytometry showed that the toxic effects of morphine on tachyzoites after 3 hours was not statistically significant (p<0.05). Also, apoptosis in infected MQs rose with a decreasing concentration of morphine. The parasitic load in MQs treated with morphine before infection was lower than that in cells treated after infection and the differences were statistically significant (p<0.01). In mice that received morphine before infection, survival rate, parasite load and the IFN-γ level were significantly lower than in mice treated after infection (p<0.01). The results of this study have shown that morphine in the pre-treatment group had higher anti-Toxoplasma activity than morphine in post-treatment in vitro and in murine model.

Highlights of human ectopic fascioliasis: a systematic review.

Fascioliasis is a tropical zoonotic disease caused by the Fasciola parasite. The adult parasite usually resides in the liver and biliary ducts; however, several cases of ectopic fascioliasis (EF) have been reported. This study is a highlight on EF according to the confirmed case reports. In a setting of systematic review, we found 25 eligible articles containing 26 confirmed cases of EF (any date until 30 November 2018), including abdominal and intestinal EF in six cases, skin and subcutaneous tissues in five cases, eye in four cases, brain and pancreas in three cases, neck and lymph node in two cases, and lung, dorsal spine, and peritoneal cavity in one case, respectively. The result indicates that fascioliasis can have diverse ectopic forms and should be more attended in the endemic regions of fascioliasis in order to distinguish from other endemic diseases.

Rhomboid antigens are promising targets in the vaccine development against Toxoplasma gondii.

Toxoplasma gondii (T. gondii) is an obligate intracellular parasite with worldwide distribution. It is estimated that near one-third of the people around the globe are latently seropositive for the parasite. Since the current common drugs are incapable in the elimination of parasites within tissue cysts, the development of an effective vaccine has high priority for researchers to limit the infection. During recent years, non-stop efforts of scientists have made great progress in the identification and development of T. gondii candidate vaccines. However, there is a lack of a commercially licensed vaccine for human application yet. Rhomboid proteases (ROMs) are a class of serine proteases that have an important role in the invasion of the parasites that can be considered as a new target for vaccine strategy. They also play critical roles in mitochondrial fusion and growth factor signaling, allowing the parasite to completely enter into the host cell. In the current review, we have summarized the recent progress regarding the development of ROM-based vaccines against acute and chronic T. gondii infection in animal models.

Recent progress in microneme-based vaccines development against Toxoplasma gondii.

Toxoplasmosis is a cosmopolitan zoonotic disease, which infect several warm-blooded mammals. More than one-third of the human population are seropositive worldwide. Due to the high seroprevalence of Toxoplasma gondii infection worldwide, the resulting clinical, mental, and economical complications, as well as incapability of current drugs in the elimination of parasites within tissue cysts, the development of a vaccine against T. gondii would be critical. In the past decades, valuable advances have been achieved in order to identification of vaccine candidates against T. gondii infection. Microneme proteins (MICs) secreted by the micronemes play a critical role in the initial stages of host cell invasion by parasites. In this review, we have summarized the recent progress for MIC-based vaccines development, such as DNA vaccines, recombinant protein vaccines, vaccines based on live-attenuated vectors, and prime-boost strategy in different mouse models. In conclusion, the use of live-attenuated vectors as vehicles to deliver and express the target gene and prime-boost regimens showed excellent outcomes in the development of vaccines against toxoplasmosis, which need more attention in the future studies.

Comparison of the cellular and biochemical properties of Plasmodium falciparum choline and ethanolamine kinases.

The proliferation of the malaria-causing parasite Plasmodium falciparum within the erythrocyte is concomitant with massive phosphatidylcholine and phosphatidylethanolamine biosynthesis. Based on pharmacological and genetic data, de novo biosynthesis pathways of both phospholipids appear to be essential for parasite survival. The present study characterizes PfCK (P. falciparum choline kinase) and PfEK (P. falciparum ethanolamine kinase), which catalyse the first enzymatic steps of these essential metabolic pathways. Recombinant PfCK and PfEK were expressed as His6-tagged fusion proteins from overexpressing Escherichia coli strains, then purified to homogeneity and characterized. Using murine polyclonal antibodies against recombinant kinases, PfCK and PfEK were shown to be localized within the parasite cytoplasm. Protein expression levels increased during erythrocytic development. PfCK and PfEK appeared to be specific to their respective substrates and followed Michaelis-Menten kinetics. The Km value of PfCK for choline was 135.3+/-15.5 microM. PfCK was also able to phosphorylate ethanolamine with a very low affinity. PfEK was found to be an ethanolamine-specific kinase (Km=475.7+/-80.2 microM for ethanolamine). The quaternary ammonium compound hemicholinium-3 and an ethanolamine analogue, 2-amino-1-butanol, selectively inhibited PfCK or PfEK. In contrast, the bis-thiazolium compound T3, which was designed as a choline analogue and is currently in clinical trials for antimalarial treatment, affected PfCK and PfEK activities similarly. Inhibition exerted by T3 was competitive for both PfCK and PfEK and correlated with the impairment of cellular phosphatidylcholine biosynthesis. Comparative analyses of sequences and structures for both kinase types gave insights into their specific inhibition profiles and into the dual capacity of T3 to inhibit both PfCK and PfEK.

A 24 bp cis-acting element essential for the transcriptional activity of Plasmodium falciparum CDP-diacylglycerol synthase gene promoter.

CDP-diacylglycerol synthase (CDS) is a key rate-limiting enzyme in the phospholipid metabolism of Plasmodium falciparum, converting phosphatidic acid to CDP-diacylglycerol. The CDS gene is predominantly expressed in the mature intraerythrocytic stages. Consequently, we physically and functionally characterized the CDS gene promoter. The mRNA transcription initiation site was mapped 121 bp upstream of the CDS gene translation start site. A 1909 bp 5' upstream sequence was isolated and found to be transcriptionally active thus constituting a functional CDS promoter. Mapping of this promoter identified a 44 bp cis-acting sequence, located between -1640 and -1596 bp upstream of the ATG codon, essential for efficient transcriptional activity. This 44 bp sequence binds specifically to nuclear factors from trophozoite stage parasites. We further showed that a 24 bp element, lying within the 44 bp sequence, mediates the specific binding to nuclear proteins and shows no significant homology to known eukaryotic DNA consensus sequence elements that bind transcription factors. The deletion of the 24 bp element abrogated promoter activity, indicating that this cis-acting sequence element is essential for efficient transcription of the CDS gene.