Identification and genotyping of Echinococcus granulosus from human clinical samples in Guilan province, north of Iran.

Cystic echinococcosis (CE) is a significant health problem in both human and veterinary medicine. It is caused by the tapeworm Echinococcus granulosus (E. granulosus). The objective of this study was to investigate molecular diversity of E. granulosus from the paraffin-embedded human (FFPE) tissue samples using sequencing of mitochondrial genes. Thirty-five FFPE tissue samples were collected from different regions of Guilan province, north of Iran. Demographic data were recorded using a questionnaire. Five sections (1 mm) of the tissue were prepared and deparaffined using xylene and ethanol methods. Molecular analysis was performed using the Nad1 and Cox1 genes using PCR and DNA sequencing. Totally, 25 cases (71.43%) were women and 10 cases (28.57%) were men. The most affected age group was 21-30 yr old. The most of cysts were isolated from the liver (n = 19; 54.29%) and others in the lung (n = 16; 45.71%). The Cox1 and Nad1 genes were successfully amplified in 16 (45.71%) and 12 (34.28%) DNA samples from FFPE tissue. Sequencing analysis revealed that all samples were E. granulosus sensu stricto complex (G1 and G3). In this study, E. granulosus sensu stricto complex G1 and G3 were identified in human hydatid cysts and showed the presence of sheep/dog cycle in human infection. This finding confirmed and completed previous studies on the geospatial distribution of E. granulosus sensu stricto complex G1 and G3 in the southern and coastal areas of the Caspian Sea region.

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.

Global epidemiology and spatial distribution of Toxoplasma gondii in goats: Protocol for a systematic review and Bayesian hierarchical meta-analysis.

BACKGROUND: Toxoplasma gondii, a cosmopolitan protozoan parasite causes toxoplasmosis in humans and many species of domestic and wild animals. T. gondii instigates significant economic losses in sheep and goat farming industry and can lead to abortion, stillbirth, congenital malformations and neonatal losses. The objective of this protocol is to evaluate worldwide seroprevalence of T. gondii exposure in goats using Bayesian hierarchical meta-analysis and geographic information system (GIS). METHODS: A comprehensive literature search will be conducted using search engines, including Web of Science, ScienceDirect, Scopus, PubMed, ProQuest, EMBASE, PROSPERO Register and, Google Scholar without date and language restrictions. The authors search for cross-sectional studies that determine the seroprevalence of T. gondii in goats. Two reviewers will independently screen, selected studies; also, they will extract data, and assess the risk of bias. In case(s) of disagreement, a consensus will be reached with the help of a third author. The Bayesian hierarchical meta-analysis will use to estimate country and worldwide true seroprevalence of T. gondii, which is consist of the sensitivity and specificity of the applied serological assays. The obtained data will be used to identify country-level risk factors associated with T. gondii exposure using GIS in the ArcGIS software. DISCUSSION: The systematic review produced from this protocol will provide the true prevalence rate and spatial distribution T. gondii exposure in goats both regionally and globally using Bayesian hierarchical and GIS analysis. SYSTEMATIC REVIEW REGISTRATION: PROSPERO registration number: CRD42020107928.

In Silico Vaccine Design and Expression of the Multi-Component Protein Candidate against the Toxoplasma gondii Parasite from MIC13, GRA1, and SAG1 Antigens.

Background: We aimed to design a B and T cell recombinant protein vaccine of Toxoplasma gondii with in silico approach. MIC13 plays an important role in spreading the parasite in the host body. GRA1 causes the persistence of the parasite in the parasitophorous vacuole. SAG1 plays a role in host-cell adhesion and cell invasion. Methods: Amino acid positions 73-272 from MIC13, 71-190 from GRA1, and 101-300 from SAG1 were selected and joined with linker A(EAAAK)A. The structures, antigenicity, allergenicity, physicochemical properties, as well as codon optimization and mRNA structure of this recombinant protein called MGS1, were predicted using bioinformatics servers. The designed structure was synthesized and then cloned in pET28a (+) plasmid and transformed into Escherichia coli BL21. Results: The number of amino acids in this antigen was 555, and its antigenicity was estimated to be 0.6340. SDS-PAGE and Western blotting confirmed gene expression and successful production of the protein with a molecular weight of 59.56kDa. This protein will be used in our future studies as an anti-Toxoplasma vaccine candidate in animal models. Conclusion: In silico methods are efficient for understanding information about proteins, selecting immunogenic epitopes, and finally producing recombinant proteins, as well as reducing the time and cost of vaccine design.

Global status of intestinal parasitic infections among diabetic patients: A systematic review and meta-analysis.

Intestinal parasitic infections (IPIs) have been identified as a disease agent responsible for infections in immunocompromised patients such as diabetics. We searched six electronic databases and reviewed 38 related studies using the following keywords alone or in combination: "intestinal parasites", "diabetes", "immunocompromised", "prevalence", and "human."  The pooled prevalence of IPIs in diabetic patients was 24.4% worldwide. These patients with IPIs are advised to go to health centers and perform the relevant checkups with the advent of the first symptoms of the disease, such as diarrhea and abdominal pain. Moreover, early diagnosis and treatment of IPIs in diabetic patients are highly recommended to maintain quality of life.

The Larval Stages of Echinostoma spp. in Freshwater Snails as the First and Second Intermediate Hosts in Gilan and Mazandaran Provinces, Northern Iran.

Background: Identification of the larval stages of Echinostoma spp. in freshwater snails is an essential guide to continue monitoring the possibility of their transmission and the potential of echinostomiasis in areas where trematodes are the primary agent of parasitic diseases. The aim of this study was investigate Echinostoma using morphological and molecular techniques. Methods: The study was conducted in Gilan and Mazandaran Provinces, northern Iran, from April 2019 to October 2021. Overall, 5300 freshwater snails were randomly collected and were identified using external shell morphology. Meanwhile, snails infected with trematodes were studied via shedding and dissecting methods. Larvae stages of Echinostoma were identified and the genomic DNA of the samples was extracted. The PCR amplification of the ITSI gene was carried out for 17 isolates and products were sequenced. Seven sequences were deposited in GenBank. Results: Totally, 3.5% of snails containing three species (Stagnicola sp., Radix sp. and Planorbis sp.) were infected with two types of cercaria, E. revolutum with 37 and Echinostoma sp. with 45 spines in the collar. Moreover, 35% of the snails were infected with Echinostoma spp. metacercaria. Phylogenetic analysis illustrated that isolates were included in two ITSI haplogroups. Conclusion: Results showed the potential hazard of a zoonotic parasite as Echinostoma in northern Iran. The potential of disease environmental relationship investigation and resource control optimization is necessary for effective disease prevention and health management.

Microscopic and Molecular Investigation of Intestinal Microsporidia in HIV + /AIDS and Cancer Patients Undergoing Chemotherapy in Mazandaran Province, North of Iran.

BACKGROUND: The aim of this study was to identify Enterocytozoon bieneusi and Encephalitozoon spp. in fecal samples of HIV + /AIDS and cancer patients undergoing chemotherapy, and comparing the results to healthy individuals in Mazandaran province, north of Iran. METHODS: Stool samples were collected from 50 HIV + /AIDS patients, 50 cancer patients, and 50 healthy samples referred to medical centers in north of Iran. Stool samples were kept in 2.5% potassium dichromate at 4 °C, and stained by modified trichrome for light microscopy examination. The multiplex/nested-PCR targeted the small subunit ribosomal RNA (SSU rRNA) gene. To characterize genotypes, the nested PCR products sequenced by Bioneer Company and was subjected to phylogenetic analyses. RESULTS: Ten of 50 samples (20%) of HIV + /AIDS patients, 5 of 50 samples (10%) of cancer patients, and 1 of healthy individuals (2%) were microscopically positive. From 50 HIV + / AIDS patients, E. bieneusi and Encephalitozoon spp. were detected in 10 (20%) and 6 (12%) cases, respectively. Furthermore, among cancer patients, 7 (14%) and 2 (4%) cases were E. bieneusi and Encephalitozoon spp., respectively. Out of 50 samples of healthy individuals, only 3 (6%) cases of E. bieneusi were observed. The genotypes D and M were detected among positive samples of E. bieneusi. CONCLUSIONS: E. bieneusi and then Encephalitozoon spp. are common intestinal microsporidia in HIV + /AIDS patients and cancer patients undergoing chemotherapy in Mazandaran province. E. bieneusi genotype D seems to be the predominant genotype in Mazandaran province. Due to the considerable prevalence of intestinal microsporidia, physicians are advised to pay more attention to this opportunistic infection in high-risk groups.

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.

Possible association between Toxoplasma gondii infection and sleep disorders: A systematic review.

BACKGROUND: Few studies investigated the relationship between toxoplasmosis and sleep disorders. Sleep disorders can lead to suicidal behavior and traffic accidents. Therefore, the purpose of this study is to collect information and investigate the possible relationship between Toxoplasma gondii (T. gondii) and sleep disorders. METHODS: To achieve the goal of the present study, five English databases (PubMed, ScienceDirect, Web of Science, Scopus, and ProQuest) were systematically searched for related studies from their inception until October 18, 2022. The obtained articles were screened based on the title, abstract and full text. Then, the quality of the papers investigating the relationship between toxoplasmosis and sleep disorders was evaluated, and finally, the data from the relevant studies were extracted in a Microsoft Excel data sheet. RESULTS: Eight articles (4 case-control and 4 cross-sectional studies) were entered in this systematic review containing 926 patients with sleep disorders and 1877 people without sleep disorders in casecontrol studies, out of which 212 (22.89%) and 392 (20.88%) individuals were positive for anti-T. gondii IgG antibody using different serological methods. Also, 2885 people with sleep disorders were investigated for anti-T. gondii IgG antibody in cross-sectional studies, out of which 1559 (54.03%) cases were positive. CONCLUSION: The results of this study suggest that T. gondii infection may be a risk factor for sleep disorders. However, the number of related studies is small, and there are contradictions in the findings of these articles. Therefore, further studies are necessary to clarify the possible association between T. gondii infections and sleep disorders.

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.

Prevalence of Ecto and Gastrointestinal Parasites of Rattus rattus in Mazandaran Province, North of Iran

Objective: Rodents act as reservoir hosts and are an important potential source for many zoonotic pathogens such as parasites, which pose a public health risk to humans. Therefore, it is necessary to investigate the prevalence of parasites among rodents. Methods: A total of 118 Rattus rattus were captured in Mazandaran province, north of Iran, using snap live traps. Various samples were collected from feces and each rat was combed with a fine-tooth comb to extricate any ectoparasite. Fecal specimens were examined by direct wet mounting, formalin-ether concentration, modified acid-fast, and trichrome staining methods. Results: The overall prevalence of gastrointestinal parasites in the examined rats was 75.4%. Cryptosporidium spp. (30.5%) were the most prevalent protozoan, followed by Giardia spp. (20.3%), Entamoeba muris (13.5%), Trichomonas muris (10.1%), and Spironucleus muris (3.3%). Regarding helminths' eggs, Syphacia obvelata (24.5%), Hymenolepis diminuta (10.1%), and Trichuris muris (9.3%) had the highest prevalence, respectively. Furthermore, 3060 ectoparasites collected from 102 rodents were infested with lice (40% Polyplax spp.), mites (33.3%), and flea (16.1% Xenopsylla cheopis and 10.6% Xenopsylla astia). Conclusion: According to the results of this study, the prevalence of ecto and gastrointestinal parasites in the collected rats in the area being studied was remarkably high. Additionally, Rattus rattus can be considered a potential risk to human health.

Genetic diversity of Toxoplasma gondii isolates from birds in the world: A systematic review.

Toxoplasma gondii (T. gondii) is one of the most important foodborne pathogens that infects a large number of vertebrate species and has a cosmopolitan distribution. Birds as intermediate hosts are very important in the life cycle of T. gondii and they can be a main source of infection for humans and felids, as well as other animals. Most species of birds feed from the ground and are the best indicator for soil contamination with T. gondii oocysts. Hence, T. gondii strains isolated from birds can represent different genotypes circulating in the environment and their main predators and consumers. The recent systematic review tries to represent the population structure of T. gondii in birds around the world. Six English language databases were searched from 1990 to 2020 to find the related studies and overall, 1275 isolates of T. gondii were separated from the analyzed samples in birds. The results of our study revealed that atypical genotypes were predominant (58.8%, 750 out of 1275). Types II, III, and I had less frequency with prevalence rates of 23.4%, 13.8%, and 2%, respectively. No isolates of Type I were reported from Africa. Summarizing ToxoDB genotypes circulating in birds around the world manifested that ToxoDB #2 was the most common (101/875), followed by ToxoDB #1 (80/875), and #3 (63/875). Totally, the results of our review represented the high genetic diversity of T. gondii with circulating non-clonal strains in birds from South and North America, while clonal parasites with low genetic diversity were predominant in Europe, Asia, and Africa.

Identification and multilocus genotyping of Toxoplasma gondii isolates from congenital infection in north of Iran.

Congenital toxoplasmosis can cause severe consequences in the fetus, such as spontaneous abortion which is affected by parasite strain. Also, recent studies revealed the high genetic diversity of Toxoplasma gondii. This study aims to investigate the serological status of T. gondii in pregnant women, multilocus genotyping in aborted fetuses' tissue, and archived formalin-fixed paraffin-embedded placenta. This study was performed on 100 pregnant women with spontaneous abortion and their aborted fetuses, and 250 of the archived placentae in Iran. The blood and tissue were examined for seroprevalence and genotype determination of T. gondii using ELISA and multilocus nested-PCR-RFLP, respectively. Anti-T. gondii IgG and IgM were detected in 68 samples (68%) and 1 (1%) out of 100 serums. Toxoplasma DNA was identified in 1 (1%) aborted fetuses' tissue and 32 (12.8%) placenta samples. Overall, ten positive DNA samples were successfully genotyped, and five genotypes were recognized (ToxoDB#1, #2, #10, #27, and #48). The obtained results indicated congenital toxoplasmosis is a severe risk in this region. As type I is highly pathogen and can lead to severe complications, the prevention of the infection should be considered in seronegative pregnant women.

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.

Effects of diverse Types of Toxoplasma gondii on the outcome of Alzheimer's disease in the rat model.

Toxoplasma gondii has lifelong persistence in the brain and its cysts can affect gene expression and change diverse biological functions of neurons. Many studies indicated T. gondii infection as a risk factor for the development of behavioral changes and neurodegenerative diseases such as Alzheimer's disease (AD), although the etiopathogenetic link between them has not been exactly elucidated. The current study aimed to examine the effects of chronic toxoplasmosis infection with Types I, II, and III strains (RH, PRU, and VEG) alone and in combination on cognitive impairments and neuronal death in the Aß1-42-induced rat model of Alzheimer's disease. In the chronic toxoplasmosis phase, Alzheimer's induction was conducted by injecting Aß1-42 oligomers into the rat brain hippocampus. Behavioral tests were conducted 10 days after the AD induction. Real-time PCR was performed to evaluate T. gondii parasite burden by amplification of the B1 gene. Cytokines IL-1ß, TNF-α, and IL-10 were assayed in brain tissue supernatant using ELISA. Also, histopathological examinations were conducted to calculate inflammatory changes and neuronal death in the brain. Our findings showed that chronic toxoplasmosis infection with PRU reduces cognitive disorders, while the RH strain of T. gondii plays a destructive role and aggravates cognitive impairments in AD. Also, infection with a combination of PRU and VEG strains significantly improved spatial learning and memory impairments in Alzheimer's rat model. Histopathological findings also confirmed the results of behavioral tests, so that in AßPRU and AßPRU + VEG groups, neuronal death and infiltration of inflammatory cells were negligible and significantly less than in Alzheimer's and AßRH groups. Our findings indicate that chronic toxoplasmosis infection with PRU strain alone, also in combination with VEG strain can significantly improve cognitive disorders in AD rats, while RH strain plays a destructive role in AD pathogenesis.

Contamination of Vector Snails with the Larval Stages of Trematodes in Selected Areas in Northern Iran.

Background: Identification of freshwater snails and possible trematodes transmission sites are essential to continue monitoring the potential for disease outbreaks in areas with a history of parasitic infections. We aimed to search some areas in the margin of the Caspian Sea, northern Iran to identify the snail fauna of this area and verify the contamination of vector snails. Methods: More than 5,308 snails from 51 diverse and permanent habitats were studied from April 2019 to October 2021. Snails were collected randomly and identified using shell morphology. Trematode infection in snails was investigated by the release of cercariae and dissection methods. Results: Five families of freshwater snails including Lymnaeidae, Physidae, Planorbidae, Bithyniidae, and Viviparidae were investigated in the Caspian Sae Litoral of Iran. Physidae were found as the most prevalent snails (55.1%) followed by Lymnaeidae (29.4%). The parasitize rate was observed as 20% using releasing cercaria technique. Echinostomatoidea (31%), Schistosomatoidea (8%), and Diplostomoidea (21%), and Plagiorchioidea (40%) were seen as detected parasites. Meanwhile, 60% of the studied snails illustrated the other stages of trematodes. Conclusion: The rate of infection of snails with different cercaria in northern Iran is significant. It needs further deep studies to clarify the situation of zoonoses transmitted by snails in the region. Policy makers should pay attention more to this area in terms of monitoring the snail-transmitted diseases.

Evaluation of anti-parasitic activities of new quinolones containing nitrofuran moiety against Toxoplasma gondii.

Toxoplasmosis is a disease with a worldwide prevalence that is caused by Toxoplasma gondii. Pyrimethamine and sulfadiazine are two pharmacological agents commonly used to treat of this infection. However, they are accompanied by some side effects. Therefore, the identifying of new drugs with low toxocytosis seems to be a matter of vital importance. Quinolones are DNA replication inhibitors, exerting inhibitory effects against many pathogens, including bacteria, mycoplasma, and protozoa. Given the importance of quinolones and their efficacy, the present in vitro study was conducted to investigate the antiparasitic activities of new quinolones (NFQ-2, NFQ-5, and NFQ-6) containing nitrofuran moiety against T. gondii. To this end, Vero cells were incubated with various concentrations of new quinolones and pyrimethamine (positive control) to determine their viability. Subsequently, they were infected with T. gondii (RH strain) and then subjected to drug treatment. The obtained IC50 values were 3.60, 4.84, 5.59, 3.44 and 2.75 µg/mL for NFQ-2, NFQ-5, NFQ-6, ciprofloxacin and pyrimethamine, respectively. The CC50 values for the NFQ-2, NFQ-5, and NFQ-6 were 25.20, 29.89, and 28.43 µg/mL, indicating the selectivity indexes more than 5 for these compounds. The anti-Toxoplasma efficiency was determined by evaluating infection index, number and size of plaques, and T. gondii intracellular proliferation. As the results indicated, the administration of new quinolone derivatives resulted in the reduction of intracellular proliferation, infection index, and the number and size of plaques in comparison to uninfected treated cells (P < 0.05). The results were indicative of a considerable synergetic effect when each of the derivatives was used in combination with pyrimethamine, compared to when used alone. Based on our results, the nitrofuran-derived quinolones can be considered as new leads for the design of new anti-Toxoplasma agents.

More Than Seventy Years of Research (1948-November 2021) on Toxoplasma gondii in Iran: A Narrative Review.

In this review, we intend to provide a summary of the activities of researchers in the field of Toxoplasma gondii in Iran, during the past 70 years. Most studies have been limited to epidemiological studies (mostly using ELISA and IFA methods). Designing a standard and reliable method using the specific antigens of this parasite is essential. So far, studies in the field of drug effects have not been able to introduce an effective drug with few side effects. Various types of vaccines have been developed, such as recombinant and DNA vaccines. However, none of them had a good efficacy. The use of multi-epitope vaccines as potential vaccines against toxoplasmosis is recommended. At present, limited studies have been conducted on the patterns of transmission and genetic diversity of isolated isolates in Iran. Future research to determine the genotype of T. gondii could play an important role in the study of population structure, and biological characteristics of this parasite. It is hoped that the results of this study will help control, prevent, and reduce the burden of disease caused by this parasite.

Saliva and tear as useful tools for the diagnosis of Toxoplasma gondii in human specimens: a systematic review.

Toxoplasmosis is diagnosed by serology, mainly using invasive specimens such as serum or cerebrospinal fluid. This study aimed to investigate whether saliva and tear samples can be used instead of serum samples to diagnose Toxoplasma gondii. Five English-language databases were checked up to 2021. Other types of non-invasive samples (milk and urine) were excluded from this study. In 15 cross-sectional studies, a total of 4338 saliva samples were examined, out of which 718/3848 (18.66%), 30/200 (15%), and 60/122 (49.18%) samples were positive for anti-T. gondii IgG, IgM, and IgA antibodies, respectively. And for tear samples, a total of 723 samples in cross-sectional studies and 153 cases and 97 controls in case-control studies were included, out of which 199/538 (36.98%) and 155/247 (62.75%) samples were positive for anti-T. gondii IgG and IgA antibodies in cross-sectional studies and 86/128 (67.18%), 1/53 (1.88%), and 78/153 (50.98%) cases and 4/47 (8.51%), 0/47 (0%), and 12/97 (12.37%) controls were positive for anti-T. gondii IgG, IgM, and IgA antibodies in case-control studies, respectively. The results suggested that antibody levels in saliva and tear samples in humans could be useful for the diagnosis of toxoplasmosis, especially ocular toxoplasmosis using tear samples.

Genetic characterization of Toxoplasma gondii in meat-producing animals in Iran.

BACKGROUND: The consumption of uncooked or undercooked food from infected intermediate hosts can result in Toxoplasma gondii infection in humans. However, few studies have investigated the genetic diversity of this protozoan parasite in Iran. The aim of the present study was to genetically characterize isolates of T. gondii from intermediate host animals in Mazandaran Province, Iran. METHODS: Blood and heart tissue samples were collected from 204 ruminants, and brain tissue was collected from 335 birds. The prevalence of T. gondii infection in these samples was determined serologically using the modified agglutination test and by conventional PCR assays. Those PCR samples positive for T. gondii DNA and 13 DNA samples extracted from aborted fetuses in a previous study were genotyped with 12 genetic markers using the multilocus-nested PCR-restriction fragment length polymorphism (Mn-PCR-RFLP) technique. RESULTS: Antibodies for parasites were found in 35.7% of the ruminant (39.1% of sheep and 26.4% of goats) samples and in 51.3% of the bird (100% of geese, 52.3% of free-range chickens and 46% of ducks) samples. Molecular detection by PCR of the repetitive 529-bp DNA fragment revealed contamination of 13.2% of ruminant (14.6% of sheep and 9.4% of goats) samples and of 9.6% of bird (11.1% of chickens, 5.7% of ducks and 0% of geese samples). The results from 30 DNA samples revealed five ToxoDB (genome database for the genus Toxoplasma) PCR-RFLP genotypes, including #1 (Type II), #2 (Type III), #10 (Type I), #27 and #48, with genotype #1 the most predominant. CONCLUSIONS: As evidenced by the results of this study, ruminants and birds are infected with T. gondii in the region, suggesting that they are likely to be involved in the transmission of T. gondii to humans through meat consumption. The identification of different genotypes may suggest a higher genetic diversity of this parasite in Mazandaran, reflecting local environmental contamination. These results have important public health implications for the prevention and control strategies of infection.