Protein profile of purified plasma- and aqueous humor-derived extracellular vesicles from patients with ocular toxoplasmosis

ABSTRACT Purpose: To characterize the extracellular vesicle protein cargo in the aqueous humor and plasma of patients with ocular toxoplasmosis. Methods: Aqueous humor and plasma were collected from six patients with active ocular toxoplasmosis and six patients with cataract. Extracellular vesicles were isolated, and western blotting and mass spectrometry were performed for protein analysis. Results: All plasma samples from patients with ocular toxoplasmosis and cataract were positive for the tetraspanins CD63 and TSG101. However, the aqueous humor from patients with ocular toxoplasmosis was positive only for CD63. Sixty-seven new unreported proteins were identified in the aqueous humor and plasma of patients with the ocular toxoplasmosis and cataract. Of the 67 proteins, 10 and 7 were found only in the cataract and ocular toxoplasmosis groups, respectively. In general, these proteins were involved in immune system activation and retina homeostasis and were related to infections and retina-associated diseases. Conclusion: The distinct protein signatures between ocular toxoplasmosis and cataract may be helpful in the differential diagnosis of ocular toxoplasmosis. However, more studies are needed to better understand the role of these proteins in the pathogenesis of ocular toxoplasmosis.

Thrombotic Events in a Patient With Acute Toxoplasmosis.

The etiology of venous thromboembolism is multifactorial, with causes being classified as either provoked or unprovoked, making it difficult to attribute a single factor as the cause of the thromboembolic event in most cases. The relationship between inflammation and thrombotic phenomena is well established. Here, we describe the uncommon occurrence of thromboses in a previously healthy patient with acute toxoplasmosis. The patient initially presented with fatigue, abdominal pain, fever and dyspnea. The diagnosis was confirmed through toxoplasmosis serology in a set of admission laboratory tests, and further imaging studies revealed the presence of pulmonary embolism and portal vein thrombosis. The patient was treated with anticoagulants and sulfamethoxazole-trimethoprim, showing improvement in the following days. This case highlights the importance of considering infectious diseases, such as toxoplasmosis, in the differential diagnosis of thrombosis, even in previously healthy individuals. To our knowledge, this is the second reported case of this association in Brazil.

Inflammatory pathways of Toxoplasmagondii infection in pregnancy.

Toxoplasma gondii (T. gondii), an obligate intracellular parasite, is considered as an opportunistic infection and causes toxoplasmosis in humans and animals. Congenital toxoplasmosis can influence pregnancy and cause mild to severe consequences for the fetal and neonatal. During early T. gondii infection, neutrophils as the most abundant white blood cells provide a front line of defense mechanism against infection. The activated dendritic cells are then responsible for initiating an inflammatory response via T-helper 1 (Th1) cells. As part of its robust immune response, the infected host cells produce interferon (IFN-γ). IFN-γ inhibits T. gondii replication and promotes its transformation from an active form to tissue cysts. Although anti- T. gondii antibodies play an important role in infection control, T-helper 2 (Th2) immune response, can facilitate the growth and proliferation of T. gondii in the host cell. In pregnant women infected with T. gondii, the expression of cytokines may vary and in response diverse outcomes are expected. Cytokine profiles serve as valuable indicators for estimating the patho-immunological effects of T. gondii infection. This demonstrates the intricate relationship between pro-inflammatory and anti-inflammatory cytokines, as well as their influence on the various pregnancy outcomes in T. gondii infection.

Effectiveness of Miltefosine Nanoemulsion Concerning the Treatment of Acute and Chronic Toxoplasmosis: In Vivo Study.

Background: Toxoplasma gondii is one of the most common parasites worldwide. It is of great importance to identify new potential drugs that are effective and less harmful in pregnant women and newborns. We investigated nanoemulsion miltefosine (NEM) in treating experimental acute and chronic toxoplasmosis. Methods: A combination of triacetin, Tween 80, and ethanol (1:2) was used for the development of NEM formulations. The size of NEM was calculated to be 17.463 nm by DLS and TEM. To investigate the performance of miltefosine (MLF), NEM, sulfadiazine (SDZ), and pyrimethamine (PYR) (positive control) in vivo, acute toxoplasmosis was induced in mice by an intraperitoneal injection of RH strain tachyzoites. After five days, the mice were examined for the number and condition of tachyzoites and histopathological changes in the liver and spleen. Chronic toxoplasmosis was investigated in rats and the number and size of brain cysts along with histopathological changes were assessed in different groups. Results: The results of the in vivo assessment of drugs in acute toxoplasmosis showed the following order regarding a decrease in the number of tachyzoites and an increase in survival rate: SDZ&PYR > NEM > MLF. The effects of drugs on chronic toxoplasmosis showed a significant effect of NEM (50%) on reducing the number of cysts compared to SDZ&PYR (10%) and MLF (12%) and reducing the size of NEM brain cysts (21%) compared to SDZ&PYR (5 %) and MLF (8%). Conclusion: Increasing the penetration of NEM through the blood-brain barrier (BBB) and subsequently reducing the number and size of T. gondii tissue cysts is a promising new drug in treating chronic toxoplasmosis.

Detection of Congenital Toxoplasmosis and Cytomegalovirus Infections Using Paired Sample Serodiagnosis from Suspected Cases at a Tertiary Teaching Hospital in Malaysia.

Background: Congenital toxoplasmosis and congenital cytomegalovirus (cCMV) infections are noteworthy in Malaysia and can cause serious health problems in neonates. The prompt and effective detection and treatment related to both illnesses may mitigate the possibility of adverse consequences from both infections. Methods: A total of 219 neonates with suspected clinical indications of congenital toxoplasmosis and/or cCMV infections from January 2022 to December 2022 were enrolled. The first samples for IgM and IgG antibodies were screened by electrochemiluminescence immunoassay. For positive results indicative of congenital toxoplasmosis and cCMV infections, second serum samples were requested and tested within a period of 2-4 weeks after testing the first sample. Results: From the 219 first serum samples, the overall seroprevalence of congenital toxoplasmosis antibodies in suspected cases was 53%; meanwhile, the overall seroprevalence of cCMV in the suspected cases was 98.6%. The results of the paired serum sample collected for investigating congenital toxoplasmosis cases revealed that 47% of the cases presented no serological evidence of exposure while the remaining 53% of cases might have acquired passive immunity from the mother. For cCMV, the number of cases with no serological evidence of exposure was 1.4%, whereas acute infection was 1.8% and possible passive immunity from the mother represented 96.8%. Conclusion: This study found a high seroprevalence of congenital toxoplasmosis and cCMV infections, probably because they are suspected cases. This study also indicates that using paired sample analysis in the categorisation of cases can aid in accurate diagnosis and more effective treatment.

The quality and characteristics of bovine sperm are compromised by Toxoplasma gondii antigens, impacting in in vitro bull fertility.

Studies in various species have demonstrated different results on the effects of T. gondii infection on sperm quality. It has also been demonstrated that in some stages of the disease, there is elimination of cellular debris or even the intact parasite in the semen. The present work aimed to evaluate the in vitro effects of the presence of soluble T. gondii antigens in bovine semen on sperm integrity. The spermatozoa were treated with T. gondii antigens in double serial dilutions classified as high, medium and low doses (8, 4, 2 µg/ml) in "TALP-Sperm" and "TALP-Fert" media. The results showed that T. gondii antigens affect sperm motility and mitochondrial activity, and cause changes in sperm chromatin integrity, as well as damage to the sperm membrane and acrosome. Finally, spermatozoa treated with T. gondii antigens were evaluated in the in vitro production of embryos (IVEP). The use of semen contaminated with antigens in IVEP routines did not lead to a decrease in the fertilization of oocytes, as sperm undergo selection before in vitro fertilization, which eliminates the most altered sperm. However, early embryonic development was affected, probably by structural changes that were not eliminated in the selection process. The results demonstrated that the presence of soluble T. gondii antigens in bovine semen alters sperm integrity and vital characteristics for the fertilization process and embryonic development and therefore causes fertility problems in males.

Seropositivity for Toxoplasma gondii in cats with clinical signs and living in households with women of childbearing age.

Toxoplasmosis caused by Toxoplasma gondii is a protozoal zoonosis with high sanitary risk for pregnant women and immunocompromised people. Felids, including domestic cats, are the only definitive hosts of T. gondii. They shed oocysts which, in the environment, become infectious for a wide range of animals, including humans, acting as intermediate hosts. This study evaluated the frequency of acute toxoplasmosis in domestic cats with compatible clinical signs and living in households with women of childbearing age. Individual serum samples were collected from 150 cats and analyzed for IgM and IgG against T. gondii. Statistical analyses were performed to evaluate associations between seropositivity and potential risk factors. Overall, 31 cats (20.7 %) were seropositive for anti-T. gondii antibodies, i.e. 9 (6.0 %) for IgM, 17 (11.3 %) for IgG and 5 (3.3 %) for both. The cats showed different combinations of clinical pictures. The following statistically significant associations were found: male sex and positivity for IgM and/or IgG (p=0.0248; OR= 0.3537; 95 % CI= 0.1528-0.8675), presence of 2 or more clinical signs and positivity to IgM only (p=0.0003; OR= +infinity; 95 % CI= 3.924 to +infinity), presence of either neurological signs (p=0.0025; OR= 13.30; 95 % CI= 3.409-61.39) or ocular signs (p=0.0228; OR= 5.835; 95 % CI= 1.631-22.37) and positivity to IgM only, presence of gastrointestinal signs and positivity to IgG only (p=0.0083; OR= 5.508; 95 % CI= 1.503-18.54). Male sex also resulted a possible risk factor in the binomial logistic regression (p= 0.011; OR= 3.336; 95 % CI= 1.131-8.44). These results indicate that cats living with women of childbearing age are at risk of infection with T. gondii. The presence of certain clinical signs can be helpful in identifying recent and/or current infections using laboratory analyses. Awareness on toxoplasmosis should be kept high to protect animal and public health.

Anti-Toxoplasma gondii effects of XYP1-derived peptides and regulatory mechanisms of XYP1.

BACKGROUND: Toxoplasmosis, caused by Toxoplasma gondii , poses serious health issues for humans and animals. Individuals with impaired immune systems are more susceptible to severe toxoplasmosis. Pregnant women infected by T. gondii can face the possibility of birth defects and miscarriages. While pyrimethamine and sulfadiazine are commonly used drugs in clinical practice, concerns over their side effects and resistance are on the rise. A spider peptide XYP1 isolated from Lycosa coelestis had potent anti-T. gondii effects, but it had a high synthesis cost and strong cytotoxicity. METHODS: This study intended to modify XYP1 for producing derived peptides via amino acid truncation and substitution. The anti-T. gondii effect was evaluated by trypan blue staining assay and killing experiment of RH strain tachyzoites. The CCK8 and hemolysis assays were used to compare their safeties. The morphological changes of T. gondii were observed by scanning electron microscope and transmission electron microscope. In addition, the mechanism of XYP1 against T. gondii through RNA-sequencing was further explored. RESULTS: In vivo and in vitro experiments revealed that XYP1-18 and XYP1-18-1 had excellent anti-T. gondii activity with lower cytotoxicity and hemolysis activity than XYP1. XYP1, XYP1-18, and XYP1-18-1 were able to disrupt the surface membrane integrity of T. gondii tachyzoites, forming pores and causing the disruption of organelles. Furthermore, RNA-sequencing analysis indicated that XYP1 could stimulate the host immune response to effectively eliminate T. gondii and lessen the host's inflammatory reaction. CONCLUSIONS: XYP1-18 had lower cytotoxicity and hemolysis activity than XYP1, as well as significantly extending the survival time of the mice. XYP1 played a role in host inflammation and immune responses, revealing its potential mechanism. Our research provided valuable insights into the development and application of peptide-based drugs, offering novel strategies and directions for treating toxoplasmosis.

Functional Characterization of 11 Tentative Microneme Proteins in Type I RH Strain of Toxoplasma gondii Using the CRISPR-Cas9 System.

Toxoplasma gondii, a pathogenic apicomplexan parasite, infects approximately one third of the world's population and poses a serious threat to global public health. Microneme proteins (MICs) secreted by the microneme, an apical secretory organelle of T. gondii, play important roles in the invasion, motility, and intracellular survival of T. gondii. In this study, we selected 11 genes of interest (GOIs) of T. gondii, tentative MICs predicted to be localized in micronemes, and we used the CRISPR-Cas9 system to construct epitope tagging strains and gene knockout strains to explore the localization and function of these 11 tentative MICs. Immunofluorescence assay showed that nine tentative MICs (TGME49_243930, TGME49_200270, TGME49_273320, TGME49_287040, TGME49_261710, TGME49_205680, TGME49_304490, TGME49_245485, and TGME49_224620) were localized or partially localized in the microneme, consistent with the prediction. However, TGME49_272380 and TGME49_243790 showed different localizations from the prediction, being localized in the endoplasmic reticulum and the dense granule, respectively. Further functional characterization of the 11 RHΔGOI strains revealed that deletion of these 11 GOIs had no significant effect on plaque formation, intracellular replication, egress, invasion ability, and virulence of T. gondii. Although these 11 GOIs are not essential genes for the growth and virulence of tachyzoites of type I RH strain, they may have potential roles in other developmental stages or other genotypes of T. gondii. Thus, further research should be performed to explore the possible role of the nine mics and the other two GOIs in other life cycle stages and other genotypes of T. gondii.

Inverted U-shaped relationship between serum 25-hydroxyvitamin D concentrations and Toxoplasma gondii infection: a cross-sectional study.

Background: Toxoplasma gondii (T. gondii) is a widespread zoonotic parasite transmitted through contaminated food or water. It poses a significant public health threat, especially to pregnant women and immunocompromised individuals. 25-Hydroxyvitamin D [25(OH)D] plays a critical role in regulating both innate and adaptive immune responses, particularly in its anti-infective capacity. However, the relationship between serum 25(OH)D concentrations and T. gondii infection remains uncertain. Methods: We analyzed the data from the National Health and Nutrition Examination Survey (NHANES) spanning 2009-2014 to explore the association between serum 25(OH)D concentrations and T. gondii infection. Extensive demographic, comorbidity, and dietary data were collected. The status of T. gondii infection was determined using serum anti-IgG antibodies. Serum 25(OH)D levels were measured using ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). In addition, weighted logistic regression and restricted cubic spline analyses were performed. Results: Our analysis included 10,157 participants (mean [SE] age, 45.38 [0.39] years; 49.73% female) who met the inclusion criteria. Serum 25(OH)D levels were categorized into quintiles, with the second quintile serving as the reference group. The final model, adjusted for age, sex, race, education level, poverty income ratio, body mass index, smoking status, hypertension, diabetes, chronic kidney disease, depression, physical activity, alcohol intake, seasonal testing, and dietary vitamin D, revealed the following adjusted odds ratios (ORs) for the quintiles: 0.75 (95% confidence interval [CI]: 0.60-0.93) for the first, 0.87 (95% CI: 0.69-1.10) for the third, 0.75 (95% CI: 0.58-0.95) for the fourth, and 0.66 (95% CI: 0.49-0.91) for the fifth. Additionally, a restricted cubic spline analysis revealed an inverted U-shaped relationship between serum 25(OH)D and T. gondii infection, with an inflection point at approximately 51.29 nmol/L. Odds ratios to the left and right of the inflection point were 1.17 (95% CI: 1.03-1.32) and 0.94 (95% CI, 0.90-0.98) per 10 nmol/L, respectively. Conclusion: Our study uncovers an inverted U-shaped relationship between serum 25(OH)D concentrations and T. gondii infection, with an inflection point around 51.29 nmol/L.

Inflammatory response and parasite regulation in acute toxoplasmosis: the role of P2X7 receptor in controlling virulent atypical genotype strain of Toxoplasma gondii.

Toxoplasmosis is a globally significant disease that poses a severe threat to immunocompromised individuals, especially in Brazil, where a high prevalence of virulent and atypical strains of Toxoplasma gondii is observed. In 1998, the EGS strain, exhibiting a unique infection phenotype, was isolated in Brazil, adding to the complexity of strain diversity. The P2X7 receptor is critical in inflammation and controlling intracellular microorganisms such as T. gondii. However, its genetic variability can result in receptor dysfunction, potentially worsening susceptibility. This study investigates the role of the P2X7 receptor during acute infection induced by the EGS atypical strain, offering insight into the mechanisms of T. gondii infection in this context. We infected the female C57BL/6 (WT) or P2X7 knockout (P2X7-/-) by gavage. The EGS infection causes intestinal inflammation. The P2X7-/- mice presented higher parasite load in the intestine, spleen, and liver. The absence of the P2X7 receptor disrupts inflammatory cell balance by reducing NLRP3, IL-1ß, and Foxp3 expression while increasing IFN-γ expression and production in the intestine. In the liver, P2X7-/- animals demonstrate diminished inflammatory infiltrate within the portal and lobular regions concurrent with an enlargement of the spleen. In conclusion, the infection of mice with the EGS strain elicited immune alterations, leading to acute inflammation and cytokine dysregulation, while the P2X7 receptor conferred protection against parasitic proliferation across multiple organs.

Zingiber officinale Ameliorates Acute Toxoplasmosis-Induced Pathology in Mice.

BACKGROUND: Toxoplasma gondii (T. gondii) infects one third of the world's population with significant illness, mainly among immunocompromised individuals and pregnant women. Treatment options for toxoplasmosis are limited which signifies the need for novel, potent, and safe therapeutic options. The goal of this study was to assess the effectiveness of the ethanolic extract of Zingiber officinale (Z. officinale) in treating mice infected with the RH T. gondii strain. MATERIALS AND METHODS: Gas Chromatography/Mass Spectrometry (GC/MS) was used to identify components of ethanolic extract of Z. officinale. A total of 80 mice were randomly allocated into four experimental groups that contained 20 mice each. The first group was left uninfected (uninfected control), while three groups were infected with T. gondii RH virulent strain tachyzoites at 2500 tachyzoites/mouse. One infected group was left untreated (infected, untreated), whereas the other two groups were treated orally with either spiramycin (positive control) or Z. officinale ethanolic extract at doses of 200 mg/kg and 500 mg/kg, respectively for 5 days, starting the day of infection. Ten mice from each group were used to assess mice survival in different groups, whereas the other ten mice in each group were sacrificed on the 5th day post-infectin (dpi) to estimate the treatment efficacy by quantifying liver parasite load, liver function, nitric oxide (NO) production, and levels of antioxidant enzymes. Additionally, histopathological studies were performed to evaluate the therapeutic effect of Z. officinale treatment on toxoplasmosis-induced pathological alterations in liver, brain, and spleen. RESULTS: Treatment with Z. officinale ethanolic extract extended the survival of mice till 9th dpi compared to 7th dpi in infected untreated mice. Higher percentage of mice survived in Z. officinale-treated group compared to spiramycin-treatment group at different time points. Liver parasite loads were significantly lower in Z. officinale extract-treated mice and spiramycin-treated mice compared to infected untreated mice which correlated with significantly lower levels of serum liver enzymes (ALT, AST) and nitric oxide (NO), as well as significantly higher catalase (CAT) antioxidant enzyme activity. Scanning electron microscopy (SEM) examination of tachyzoites from the peritoneal fluid revealed marked damage in tachyzoites from Z. officinale-treated group compared to that from infected untreated mice. Moreover, treatment with Z. officinale ethanolic extract alleviated infection-induced pathological alterations and restored normal tissue morphology of liver, brain, and spleen. CONCLUSION: Our results demonstrated that Z. officinale treatment reduced parasite burden and reversed histopathological and biochemical alterations in acute murine toxoplasmosis. These findings support the potential utility of Z. officinale as a future effective natural therapeutic for toxoplasmosis. Further studies are needed to determine the effective active ingredient in Z. officinale extract that can be further optimized for treatment of toxoplasmosis.

Human Ocular Toxoplasmosis in Romania: History, Epidemiology, and Public Health: A Narrative Review.

Toxoplasmosis, caused by the protozoan parasite Toxoplasma gondii (T. gondii), presents a significant global health concern, particularly for immunocompromised individuals and congenitally infected newborns. Despite its widespread prevalence, there are limited data on T. gondii seroprevalence and ocular toxoplasmosis in Romania. This review aims to summarize the research accomplished on the prevalence and epidemiology of human ocular toxoplasmosis in Romania. Ocular toxoplasmosis, a leading cause of infectious posterior uveitis worldwide, involves complex interactions between host immune responses and parasite factors. Clinically, it presents as focal necrotizing retinitis, characterized by active focal retinal lesions with adjacent chorioretinal scarring, often accompanied by vitreous inflammation and anterior chamber reactions. Diagnosis relies on clinical examination supported by fundus photography, optical coherence tomography (OCT), and serological assays. The authors followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) standards, conducting a literature review on PubMed, Google Scholar, and Scopus. Our focus was on ocular toxoplasmosis in Romania, and we used keywords and specific MeSH terms. Finally, 17 articles met all the criteria, as summarized in the PRISMA diagram. This study underscores the need for improved diagnostic methods, increased research efforts, and comprehensive public health education to mitigate the burden of toxoplasmosis and ocular toxoplasmosis in Romania.

Single Cell Expression Systems for the Production of Recombinant Proteins for Immunodiagnosis and Immunoprophylaxis of Toxoplasmosis.

Toxoplasmosis represents a significant public health and veterinary concern due to its widespread distribution, zoonotic transmission, and potential for severe health impacts in susceptible individuals and animal populations. The ability to design and produce recombinant proteins with precise antigenic properties is fundamental, as they serve as tools for accurate disease detection and effective immunization strategies, contributing to improved healthcare outcomes and disease control. Most commonly, a prokaryotic expression system is employed for the production of both single antigens and multi-epitope chimeric proteins; however, the cloning strategies, bacterial strain, vector, and expression conditions vary. Moreover, literature reports show the use of alternative microbial systems such as yeast or Leishmania tarentolae. This review provides an overview of the methods and strategies employed for the production of recombinant Toxoplasma gondii antigenic proteins for the serological detection of T. gondii infection and vaccine development.

Impact of education on knowledge, attitudes, and practices for gestational toxoplasmosis.

BACKGROUND: Toxoplasmosis is potentially avoidable, treatable, and curable by simple and direct preventive measures. Knowledge, attitudes, and practices (KAP) assessments concerning gestational toxoplasmosis were evaluated in a cohort of pregnant women from Armenia-Quindío (Colombia, South America). METHODS: This cross-sectional descriptive KAP-type study was performed with informed consent between October 2021 and March 2022. The intervention involved a ten-minute talk administered by prenatal clinic nurses to pregnant women. This took place in the public health clinic RedSalud and the private clinic Happy Maternity with a post-KAP survey after pregnancy. RESULTS: The findings of the initial KAP survey revealed that approximately 42.8 % of the 250 mothers surveyed had IgG anti-T. gondii antibodies present. A strong correlation was observed between a lower frequency of antibodies and a higher level of education. Following an educational intervention, 73 seronegative women demonstrated a significant improvement in their knowledge and behavior. Among the 111 mothers who received the intervention, 42 (37 %) were followed until delivery. Unfortunately, their level of compliance with prenatal serological follow-up was lower compared to previous historical records of cohort of mothers in the same health center during pre-pandemic periods. No seroconversion occurred, although the small number of cases makes the outcome inconclusive with respect to statistical significance. CONCLUSIONS: Education plays a crucial role in imparting valuable knowledge and fostering effective practices. It holds significant potential to prevent toxoplasmosis in pregnant seronegative mothers. Prenatal check-ups have proven to be a critical determinant in leveraging the benefits of education for seronegative mothers. Reporting and observed behaviors differed, identifying areas for improvement.

Molecular detection of toxoplasmosis in wild rats using loop-mediated isothermal amplification assay.

Background and Aim: Toxoplasmosis is caused by the parasite Toxoplasma gondii. Cats are the only known hosts that excrete resistant oocysts. Wild rats serve as crucial reservoirs and intermediate hosts for T. gondii's survival and dissemination. Consuming soil and water containing oocysts can lead to illness. This study aimed to estimate the prevalence of toxoplasmosis in wild rats through molecular detection as an indicator of environmental contamination in Surabaya. Materials and Methods: One hundred rats were collected from the three areas (housing, dense settlements, and traditional markets) and distributed into the five zones: West, East, Central, North, and South of Surabaya. Brain tissue samples were extracted using a Geneaid™ (New Taipei City, Taiwan) DNA isolation kit and analyzed through the loop-mediated isothermal amplification (LAMP) method. Results: The study analyzed brain tissue from 100 wild rats, consisting of 77 Rattus tanezumi and 33 Rattus norvegicus, displaying 30% LAMP positivity. The study revealed that 30% (30/100) of wild rats tested were infected with T. gondii. The molecular prevalence rate in male rats was 32.35% (22/68), compared to females with 25% (8/32). 41.9% of the housing population, 33.3% of traditional markets, and 22.6% of dense settlements had the highest molecular prevalence. The high positive molecular rate at the trapping site can be attributed to cats and dense populations. Conclusion: Thirty percentage wild rats were tested positive for toxoplasmosis in Surabaya, East Java, Indonesia using LAMP method. Implementing strict control and monitoring is crucial in preventing the transmission of diseases from wild rats to humans. It is necessary to carry out further research related to genetic analysis of T. gondii to determine the type of T. gondii that infects animals and humans in Surabaya through bioassay and molecular test.

Seroprevalence of IgG and IgM Antibodies against Toxoplasma gondii in Dogs in Greece.

Toxoplasmosis, caused by the protozoan Toxoplasma gondii, is a zoonotic disease that affects various animal species, including dogs, that can serve as sentinels for indirectly estimating the environmental contamination. The current study aimed to determine the seroprevalence of T. gondii IgG and IgM antibodies in dogs across different regions of Greece and assess their living area as a potential risk factor. In total, 1282 blood samples were collected from dogs in urban and rural areas of Greece, including Attica and Thessaloniki. Serum samples were tested for T. gondii-specific IgG and IgM antibodies using an indirect immunofluorescent antibody test. A chi-square test was performed to assess the association between seropositivity for T. gondii and geographical location (urban/rural). The overall T. gondii seroprevalence was 47.6%, while the seroprevalence of IgG and IgM antibodies was 34.3% and 22.2%, respectively. Dogs from rural areas exhibited a significantly higher seroprevalence (53.8%) than those from urban areas (43.9%) (p < 0.001), with the estimated odds ratio being equal to 1.49 (95% CI, 1.18 to 1.65) and the relative risk increased by 22.4%. Dogs in Greece are highly exposed to T. gondii, particularly in rural areas. Measures to prevent canine infections are necessary, and basic hygiene practices, such as hand washing after petting dogs, are required to reduce human infection risk and safeguard public health.

The effect of infection with Toxoplasma gondii in inducing interferon-gamma in breast cancer patients.

Toxoplasmosis is one of the most widespread zoonotic diseases in the world. Human infection rates range from 10% to 80% in many countries. Female cancer patients receiving chemotherapy are more susceptible to developing acute forms of toxoplasmosis, which can cause brain defects, neurological damage, and encephalitis. The aim of the present study was to investigate the effect of Toxoplasma gondii infection on the induction of interferon-gamma in breast cancer patients from Iraq. This descriptive cross-sectional study was performed on women had breast cancer in Al-Haboubi Teaching Hospital in Nasiriya City-Thi-Qar Province (Iraq) during the period from January to September 2022. Approximately three ml of blood was drawn from all participants and sera were collected. The Sera were then tested for Toxoplasma IgM, IgG, and IFN-γ (Nova Tec Immunodiagnostica GmbH, Germany) using the enzyme-linked immunosorbent assay (ELISA) kits according to the protocols of the manufacturer. Before blood collection, participants completed a printed questionnaire with some demographic information, such as age and place of residence. The total number of positive T. gondii infections from breast cancer patients in the current study was 60 (85.7%). The results of sample analysis by ELISA assay showed that 85.7% and 74.2% of patients were positive for IgG and IgM, respectively. The mean IFN-γ levels in breast cancer patients with toxoplasmosis, without toxoplasmosis, and in the control group were 47.66, 0.00, and 0.57 pg/ ml, respectively. Higher IgG and interferon gamma levels were detected in the group of breast cancer patients with toxoplasmosis than in the group without toxoplasmosis. According to the ELISA findings, T. gondii was the most common parasite in female cancer patients.