anti-Toxoplasma Gondii Research Articles

Seroepidemiology of Toxoplasma gondii infection and parasite DNA in free-range chickens in Aguascalientes, Mexico

The objectives of the study were to estimate the prevalence of anti-Toxoplasma gondii antibodies in free-range chickens in Aguascalientes, Mexico, its association with certain risk factors, and the frequency ofparasite DNA in the heart. Eighty-one small rural family farms were included, and blood and heartsamples were taken from 150 clinically healthy, adult, free-range chickens. Serum samples were processed by indirect immunofluorescence antibody test considering a dilution of 1:16 as a positive reaction, while the DNA detection was done by PCR. The correlation between the presence of antibodies and the potential risk factors was estimated with logistic regression. The overall seroprevalence in the examined populations was 67%, with a range from 33% to 100% among municipalities, and 78% of the farms having at least one seropositive bird. The PCR test identified the presence of parasite DNA for one case only. Among the variables, the presence of cats and the presence of other animal species on the farm were the ones that had the highest values, while the presence of feral cats and the presence of noxious fauna were also identified as potential risk factors. The results indicate high soil contamination by T. gondii oocysts, which implies the active role of cats living on the farms.

Seroprevalence and genetic characterization of Toxoplasma gondii in domestic pigs intended for human consumption in Cuba.

Domestic pigs are considered as one of the main intermediate hosts in the zoonotic transmission of Toxoplasma gondii in many countries. Serological and molecular studies are warranted to better understand the epidemiology and transmission patterns of this parasite worldwide. To date, seroepidemiological information on T.gondii in domestic pigs in Cuba is very scarce and there are no reports of T.gondii genotypes circulating in this country. Here, we aimed to estimate the seroprevalence of T.gondii and provide genetic characterization of the strains circulating in slaughtered pigs intended for human consumption in Central Cuba. Seroprevalence was determined in 450 serum samples from slaughtered pigs in Villa Clara province using ELISA. Anti-Toxoplasma gondii IgG antibodies were detected in 100 animals (22.2%, 95% CI: 18.5-26.2). Conventional PCR of the 529-bp marker of T.gondii was performed in hearts and diaphragm tissues of all ELISA-seropositive pigs. Toxoplasma gondii DNA was detected in four animals. Further genetic characterization of the positive DNA samples was performed by multilocus PCR-RFLP and PCR-sequencing typing tools. Molecular analysis revealed four different genetic profiles that were combinations of type I, II, III and u-1 alleles, suggesting the circulation of non-clonal genotypes of T.gondii in domestic pigs in Cuba. Our results indicate that T.gondii is widely distributed in slaughtered pigs in this country, which might have important implications for public health. To the best of our knowledge, this is the first report on genetic characterization of T.gondii in Cuba. Although preliminary, the results suggest a high genetic diversity of T.gondii in the study region. Further studies based on parasite isolation are needed to definitively identify the genotypes circulating and characterize the virulence of strains detected in pigs in Cuba, and to assess the risk of zoonotic transmission from pork products in this country.

Seroprevalence of anti-Toxoplasma gondii antibodies and associated risk factors in pregnant women in Somalia

Toxoplasma gondii infection is a worldwide protozoan parasitic disease caused by the parasite Toxoplasma gondii. T. gondii infection in gestation period can lead to fetal or neonatal death, and besides congenital defects. So far, there are no data available regarding T. gondii seropositivity and associated risk factors in pregnant women in Somalia. The investigations of these aspects were done in 403 pregnant women attending the antenatal care (ANC) at Benadir hospital, in Mogadishu city. Blood samples were collected and analyzed for anti-T. gondii antibodies by enzyme-linked immunosorbent assay (ELISA). IgG seropositivity to T. gondii was 45.2% (182/403). None of the women were positive for IgM antibodies. Considering the first, second, and third trimesters of pregnancy, women IgG seropositivity was 44.2%, 26.3%, and 67.7%, respectively (p < 0.05). There was no significant association between seropositivity and age, sociodemographic features, history of abortion, presence of cats in house, contact cats feces, contact with soil, consumption of raw vegetables/meats, blood transfusion, and drinking of unboiled milk (p>0.05). Routine serological screening for T. gondii should be integrated with other ANC services to identify potential infections with the parasite.

Negative trend in seroprevalence of anti-toxoplasma gondii IgG antibodies among the general population of the province of Vojvodina, Serbia, 2008–2021

This study aimed to estimate dynamic changes in seroprevalence of Toxoplasma gondii within the general population living in the northern part of the Republic of Serbia (Province of Vojvodina) during a 14-year period. The differences in prevalence of anti-toxoplasma antibodies were analyzed in correlation with age, gender, residential area (rural/urban) and meteorological factors. In this cohort retrospective study, 24,440 subjects between 1 and 88 years old were enrolled. To determine the presence of T. gondii-specific IgM and IgG antibodies in serum samples, commercially available ELISA kits were used (Euroimmun, Luebeck, Germany). During the study period, the overall T. gondii seroprevalence was 23.5%. The seroprevalence continuously decreased over time from 31.7% in 2008 to 20.4% in 2021 (0.81% per year, p < 0.001). Approximately 2% of patients had a serologic profile positive for both anti-Toxoplasma IgG and IgM antibodies. The seroprevalence was higher (28.87%) among men compared to women (24.28%), while urban residents (24.94%) had lower seroprevalence than the rural population (28.17%). A statistically significant negative correlation (r = −0.559) was found between serologic profile of patients positive for both T. gondii IgG and IgM antibodies and the annual mean air temperature. No significant association was observed between seropositivity to T. gondii infection and examined meteorological factors. These data could be useful to national and regional health authorities to create an optimal health policy to reduce rate of T. gondii infections.

Seroprevalence of Toxoplasma gondii among hemodialysis patients: A possible link to main T-lymphocyte subsets levels and dialysis adequacy

BackgroundA high prevalence of Toxoplasma gondii infection has been reported in patients with hemodialysis (HD). However, a lack of data exists on the relationship between T lymphocyte subsets and dialysis adequacy. This study aimed to investigate the seroprevalence of Toxoplasma gondii infection among HD patients and its relation to T lymphocyte cells subsets, CD3+, CD4+, CD8+, CD4+/CD8+ ratio, and HD adequacy. MethodsThis cross-sectional comparative study was conducted on 92 subjects. Of them, 42 HD patients and 50 were control participants. Anti-Toxoplasma gondii IgG, IgM antibodies, the T lymphocyte cells subset CD3+, CD4+, CD8+, CD4+/CD8+ ratio, and adequacy of dialysis via calculation of Kt/V were detected for all subjects. ResultsThe seropositivity for anti-Toxoplasma gondii IgG antibodies was significantly higher in HD patients 66.7% (28/42) compared to 34% in controls (17/50), (p = 0.0003). The main T lymphocyte subsets was significantly lower in HD compared to controls (p < 0.05). Seropositive HD patients exhibited statistically significantly lower T lymphocyte cell subsets and CD4+/CD8+ ratio compared to seronegative patients (p < 0.05). There was a negative correlation between anti-Toxoplasma gondii IgG level and T lymphocyte subsets and the CD4/CD8+ ratio. Binary logistic regression showed that CD4+ T cell significantly predicts Toxoplasma gondii susceptibility among HD patients (p = 0.03). The mean Kt/V index is significantly lower among seropositive HD patients compared to seronegative HD patients (1.05 ± 0.46, 1.41 ± 0.53, respectively, p = 0.03) with a significant negative correlation with anti-Toxoplasma gondii IgG level (p < 0.05). ROC curve analysis showed the CD4+ T cell % had the highest AUC value among HD patients (AUC= 0.88, p < 0.001). The Kt/V value of ≤ 0.8 significantly predicted susceptibility to Toxoplasma gondii infection among HD patients (AUC = 0.68, p = 0.03). ConclusionThe current study reinforces previous reports of a high prevalence of Toxoplasma gondii infection among HD patients. CD4+ T cell and Kt/V showed a good diagnostic performance in predicting susceptibility for Toxoplasma gondii infection in HD patients. Considering the clinical consequences caused by Toxoplasma gondii infection in these patients, regular screening and adequate HD are recommended.

Inhibition of Toxoplasma gondii by 1,2,4-triazole-based compounds: marked improvement in selectivity relative to the standard therapy pyrimethamine and sulfadiazine

A safer treatment for toxoplasmosis would be achieved by improving the selectivity profile of novel chemotherapeutics compared to the standard therapy pyrimethamine (PYR) and sulfadiazine (SDZ). We previously reported on the identification of the compounds with imidazole-thiosemicarbazide scaffold as potent and selective anti-Toxoplasma gondii (T. gondii) agents. In our current research, we report on the optimisation of this chemical scaffold leading to the discovery cyclic analogue 20 b with s-triazole core structure. This compound displayed prominent CC30 to IC50 selectivity index (SI) of 70.72, making it 160-fold more selective than SDZ, 11-fold more selective than PYR, and 4-fold more selective than trimethoprim (TRI). Additionally, this compound possesses prerequisite drug-like anti-Toxoplasma properties to advance into preclinical development; it showed ability to cross the BBB, did not induce genotoxic and haemolytic changes in human cells, and as well as it was characterised by low cellular toxicity.

Seropositivity of Anti-Toxoplasma gondii Antibodies in Owners and Their Dogs Living on Island and Mainland Seashore Areas of Southern Brazil.

Although toxoplasmosis has been considered among the most neglected zoonoses worldwide, no study has focused on the frequency and associated risk factors of owners and their dogs living on an island and mainland seashore areas. Accordingly, anti-Toxoplasma gondii IgG antibodies were screened by indirect fluorescent antibody test (IFAT) in owners and dogs from three oceanic islands and two nearby mainland harbor areas, with associated risk factors for toxoplasmosis assessed by univariable and multivariable logistic regression models. Overall, anti-T. gondii seropositivity was observed in 59/328 (18.0%) owners and 66/283 (23.3%) dogs, with no statistical difference between owners (p = 0.360) and dogs (p = 0.655) from islands and mainland areas. Consumption of local water springs (p = 0.016; OR = 2.11) was an associated risk factor for T. gondii seropositivity, and owners with the habit of spring water intake were twice more likely seropositive (p = 0.014; OR = 2.14). Presence of anti-T. gondii antibodies in dogs was associated with seropositive owners (p = 0.008; OR = 2.81), household consumption of beef meat (p = 0.042; OR = 1.7) and chicken (p = 0.026; OR = 2.9). Despite being lower than the worldwide prevalence, toxoplasmosis seropositivity in owners and their dogs in southern Brazil was influenced by the positive owner, water source, and meat consumption, and not by inhabiting islands or seashore mainland areas, presence of dogs, cats, or both. In addition, drinking water quality should always be considered a critical risk factor for toxoplasmosis on islands.

Frequency of anti-Toxoplasma gondii antibodies in dogs and cats from the metropolitan region of Vitória, Espírito Santo, Brazil

Toxoplasmosis, an infection caused by Toxoplasma gondii, is widely distributed worldwide. Seroprevalence in pets is a reliable tool to determine environmental parasite presence and human risk of infection due to proximity to these animals. In this context, the frequency of anti-T. gondii IgG antibodies in sera of dogs and cats from Espirito Santo, Brazil and risk factors associated with the infection (sex, age, allocation, and breed) were assessed. Antibodies against T. gondii were detected in 39.4% (149/378) and 38.1% (142/373) of the dogs by enzyme-linked immunosorbent assay (ELISA) and by indirect immunofluorescence antibody test (IFAT), respectively. In cats, antibodies were detected in 15.2% (12/79) by ELISA and 7.6% (6/79) by IFAT. Canine infection was associated with stray origin and older ages. Feline females showed a higher chance of infection. There was an agreement between techniques in the detection of antibodies in dogs (k = 0.82) and cats (k = 0.63). These results indicate the parasite presence in the urban environment, suggesting the possibility of infection to humans and other animals. However, this risk is lower while considering pet domiciled animals with a more controlled diet and better hygiene conditions.

In vitro and in vivo anti-Toxoplasma activities of HDAC inhibitor Panobinostat on experimental acute ocular toxoplasmosis.

Ocular toxoplasmosis (OT) is retinochoroiditis caused by Toxoplasma gondii infection, which poses a huge threat to vision. However, most traditional oral drugs for this disease have multiple side effects and have difficulty crossing the blood-retinal barrier, so the new alternative strategy is required to be developed urgently. Histone deacetylases (HDAC) inhibitors, initially applied to cancer, have attracted considerable attention as potential anti-Toxoplasma gondii drugs. Here, the efficacy of a novel HDAC inhibitor, Panobinostat (LBH589), against T. gondii has been investigated. In vitro, LBH589 inhibited the proliferation and activity of T. gondii in a dose-dependent manner with low toxicity to retinal pigment epithelial (RPE) cells. In vivo, optical coherence tomography (OCT) examination and histopathological studies showed that the inflammatory cell infiltration and the damage to retinal architecture were drastically reduced in C57BL/6 mice upon treatment with intravitreal injection of LBH589. Furthermore, we have found the mRNA expression levels of inflammatory cytokines were significantly decreased in LBH589–treated group. Collectively, our study demonstrates that LBH589 holds great promise as a preclinical candidate for control and cure of ocular toxoplasmosis.

Central nervous system aspergillosis misdiagnosed as Toxoplasma gondii encephalitis in a patient with AIDS: a case report

BackgroundPatients with acquired immunodeficiency syndrome (AIDS) tend to suffer from several central nervous system (CNS) infections due to hypoimmunity. However, CNS aspergillosis (CNSAG) is extremely rare and difficult to diagnose. Thus, it is easily misdiagnosed.Case presentationWe reported a 47-year-old male AIDS patient with ghosting vision and anhidrosis on the left head and face. He was accordingly diagnosed with Toxoplasma gondii encephalitis (TE) at other hospitals, for which he received regular anti-Toxoplasma gondii and anti-human immunodeficiency virus (anti-HIV) treatment. Then, the patient was transferred to our hospital due to a lack of any improvement with the prescribed treatment. The patient's neurological examination revealed no abnormalities at admission, only a slight change in the cerebrospinal fluid. His cranial magnetic resonance imaging (MRI) revealed multiple abnormal signals in the brain parenchyma, and his blood was positive for Toxoplasma gondii IgG antibody. The initial diagnosis at our hospital was also TE. Considering the poor efficacy of anti-TE treatment, cerebrospinal fluid metagenomics next-generation sequencing (mNGS) was performed, but no pathogenic bacteria were detected. However, Aspergillus fumigatus was detected in the cerebrospinal fluid via targeted next-generation sequencing (tNGS) and bronchoalveolar alveolar lavage fluid via mNGS. The diagnosis was accordingly revised to CNSAG combined with his other clinical manifestations. After administering voriconazole antifungal therapy, the patient’s symptoms were relieved, with improved absorption of the intracranial lesions.ConclusionsThe present case experience indicates the need for clinicians to strengthen their understanding of CNSAG. Moreover, for patients with diagnostic difficulties, early mNGS and tNGS (using biological samples with only a few pathogens) are helpful for early diagnosis and treatment, potentially allowing patients to achieve favorable outcomes.

Seroprevalence of Toxoplasma gondii among ocular infection patient in Shahid Dr. Aso Hospital in Sulaimani City, Iraq

Toxoplasma gondii is an intracellular protozoan parasite that causes ocular toxoplasmosis with most complications such as developing eye lesions and impaired vision. During the period from October 2021 to April 2022, a total of 174 blood samples were taken from ocular infected patients whom visited Shahid Dr. Aso Hospital in Sulaimani City, Iraq. These samples were tested serologically for detection of anti-Toxoplasma gondii antibodies (IgG and IgM) with both imunochromatograhpy (rapid test) and ELISA. The serological test demonstrated that 63 IgG (36.2%) and 4 IgM (2.2%) were positive by rapid test imunochromatograhpy. While, 80 IgG (45.9%) and 2 IgM (1.1%) were positive by ELISA. Most of the infections (58.75%) were between 50-69 years old. All patients (80) were examined for intraocular toxoplasmosis lesion, only 10 patients (12.5%) including 2 males (2.5%) and 8 females (10%) had ocular toxoplasmosis lesions. Most of the lesions were scars (old type) while, in some patients they were active lesions. The types of lesions, numbers per each eye, lesion diameter and comparison with the IgG and IgM titers were demonstrated.

Anti-Toxoplasma gondii IgM Long Persistence: What Are the Underlying Mechanisms?

Diagnosis of Toxoplasma gondii acute infection was first attempted by detection of specific IgM antibodies, as for other infectious diseases. However, it was noted that this immunoglobulin declines slowly and may last for months or even years. Apart from the diagnostic problem imposed on clinical management, this phenomenon called our attention due to the underlying phenomena that may be causing it. We performed a systematic comparison of reports studying IgM antibody kinetics, and the data from the papers were used to construct comparative plots and other graph types. It became clear that this phenomenon is quite generalized, and it may also occur in animals. Moreover, this is not a technical issue, although some tests make more evident the prolonged IgM decay than others. We further investigated biological reasons for its occurrence, i.e., infection dynamics (micro-reactivation–encystment, reinfection and reactivation), parasite strain relevance, as well as host innate, natural B cell responses and Ig class-switch problems inflicted by the parasite. The outcomes of these inquiries are presented and discussed herein.

Expression of Interleukin-33 Gene in Hemodialysis Patients with Chronic Toxoplasmosis in Baghdad, Iraq.

Toxoplasmosis is a protozoan parasite with high distribution, leading to different abnormalities in hosts. The present study aimed to determine the distribution of toxoplasmosis in hemodialysis patients and the expression of the Interleukin (IL)-33 gene in chronic Toxoplasmosis. The present study evaluated 120 subjects, including 60 patients who were undergoing dialysis and 60 healthy samples as the control group, from the 1st February to 1st November 2021. Anti-Toxoplasma gondii IgG was detected by using the enzyme-linked immunosorbentassay(ELISA) technique and the real-time polymerase-chain-reaction (PCR) was used to perform IL-33. The results demonstrated that the highest anti-toxoplasmosis IgG antibody rate was observed in the age group 51-70 years who were undergoing dialysis, in comparison with that in the control group (P<0.05). The male patients who had anti-toxoplasmosis IgG antibodies outnumbered the healthy people (P<0.05), while the female patients did not significantly differ from the healthy group. Chronic toxoplasmosis showed a higher number according to residency (the urban and rural patients), compared to healthy people. The frequency of dialysis times per week in chronic Toxoplasmosis patients was significantly higher among the infected patients. The findings were displayed to be positive in dialysis at 2 weeks (P<0.05). The expression of the IL-33 gene was investigated in patients who were undergoing hemodialysis and in healthy controls by using real-time PCR. The findings demonstrated that there was a high Ct value for patients and controls with a high Ct value of templates, preoperational to the gene concentration. The high prevalence of toxoplasmosis in dialysis patients and the role of IL-33 in cellular immunity in these patients highlight the need to investigate the mechanisms limiting infection with intracellular protozoa.

Synthesis and evaluation of in vitro and in vivo anti-Toxoplasma gondii activity of tetraoxane-substituted ursolic acid derivatives

A series of derivatives of ursolic acid (UA) were synthesised, the anti-Toxoplasma gondii activity was tested, and the selectivity index (SI) of these compounds was calculated to determine the derivative with the best anti-Toxoplasma gondii activity. Compound A7 showed the best activity against the Toxoplasma gondii (IC50 in T. gondii infected GES-1 cells: 9.1 ± 7.2 μM), better than the lead compound UA and the positive control drug Spiramycin. Compound A7 was selected for further in vivo research: A7 was tested for its effect on the inhibition rate of tachyzoites in mice and its biochemical parameters, such as alanine aminotransferase, aspartate aminotransferase, glutathione, and malondialdehyde were determined. Compound A7 was evaluated for its anti-Toxoplasma activity and partial damage to the liver. Therefore, the results show that compound A7 could be a potential lead compound for developing a novel anti-Toxoplasma gondii molecule.

Potential risk factors for the presence of anti-Toxoplasma gondii antibodies in finishing pigs on conventional farms in the Netherlands

BackgroundThe parasite Toxoplasma gondii (T. gondii) causes a substantial human disease burden worldwide. Ingesting improperly cooked pork containing T. gondii is considered one of the major sources of human infection in Europe and North America. Consequently, control of T. gondii infections in pigs is warranted. The European Food Safety Authority advised to perform serological monitoring of pigs and to conduct farm audits for the presence of risk factors. Serological monitoring was implemented in several Dutch slaughterhouses, one to six blood samples (a total of 5134 samples) were taken from each delivery of finishing pigs and samples were tested for the presence of anti-T. gondii antibodies. Using these test results, a cross-sectional study was initiated to assess the association between the within-herd T. gondii seroprevalence and the presence of risk factors for T. gondii infections at 69 conventional finishing pig farms in the Netherlands.ResultsA multivariable model showed significant (P ≤ 0.05) association with twelve potential risk factors: type of farm, presence of dogs, presence of ruminants, use of boots, use of shower and farm clothing, mode of rodent control, bedding accessibility for rodents, presence of cats, type of drinking water, heating of the feed, use of goat whey and shielding of birds.ConclusionsSerological monitoring of finishing pigs for T. gondii in slaughterhouses can be used to identify the presence of T. gondii risk factors on Dutch conventional finishing pig farms and seems a valuable tool to guide and monitor the control of T. gondii in pork production.

Brain toxoplasmosis after HLA haploidentical transplantation for acute myeloid leukemia.

A 66-year-old man with acute myeloid leukemia with a complex karyotype achieved complete remission with idarubicin + cytarabine (Arac) (3 + 7) followed by high-dose Arac for three cycles which were followed by haploidentical stem-cell transplantation with fludarabine, intravenous busulfan, and low-dose total body irradiation. Post-transplant cyclophosphamide was used for the prevention of graft-versus-host disease (GVHD). Neutrophil engraftment was observed on day 21. On day 32, the patient developed acute gastrointestinal GVHD and was started on prednisolone (1 mg/kg). GVHD gradually improved, and prednisolone was reduced, but on day 72, the patient developed pulmonary GVHD and was treated with a high-dose corticosteroid. Around the same time, he began to experience disorientation, which worsened rapidly. His respiratory condition gradually improved with immunosuppressive drugs and artificial respiration, but his disorientation persisted. Computed tomography (CT) of the brain showed no abnormalities, and magnetic resonance imaging showed multiple high-signal findings with myelography. Examination of the cerebrospinal fluid was negative for both bacteria and viruses. Because serum toxoplasma immunoglobulin M (IgM) was elevated, that is 1.4 IU/ml (range: 0.0–0.8 IU/ml), we considered the possibility of cerebral toxoplasmosis and administered sulfamethoxazole/trimethoprim; however, there was no improvement in consciousness, and the patient died on day 99. A brain CT scan taken after his death showed multiple small calcified lesions in his brain (Figure 1A), and autopsy findings showed infarct-like necrosis (Figure 1B) and Toxoplasma cysts (bradyzoites) in the bilateral basal ganglia, cerebral cortex, and subcortical areas (Figure 1C). Immunostaining by anti-Toxoplasma gondii antibody was positive (Figure 1D). Multiple cerebral calcified lesions are a typical picture of congenital cerebral toxoplasmosis, but they are rare in cases of acquired cerebral toxoplasmosis in immunosuppressed adults. Moreover, calcification in a short period, as in this patient, has rarely been reported before. The authors received no specific funding for this work. We also thank Editage (www.editage.jp) for English language editing. The authors declare they have no conflicts of interest. Ethical approval was not sought for the present study because this study is only a single case report with complete removal of all identifying information. The patient provided consent for the publication of this case with the removal of all identifying information to remain anonymous and retain his privacy. Takafumi Tsushima designed the study, collected data, wrote the manuscript, and provided patient care. Rikako Tabata and Kentaro Narita provided patient care. Koichi Honma and Kengo Takeuchi performed pathology. Kosei Matsue supervised this paper. All authors reviewed and approved the manuscript.

Investigation of Seroprevalence of Toxoplasmosis in Horses and Donkeys in Muş Province of Turkey

The purpose of this study was to determine the seroprevalence of Toxoplasmosis in equidae in the province of Muş, Turkey. The study material consisted of 210 equidae including 159 horses and 51 donkeys in Mus province. In serum samples, anti-Toxoplasma gondii antibodies and titers were detected using Sabin Feldman Dye Test (SFDT). Seropositivity was found in 115 (54.76 %) of the 210 equidae tested in the study. The rate of seropositivity in donkeys (92.16%) was higher than the rate in horses (42.77%), and statistical significance was observed (P&lt;0.001). T. gondii antibody was detected in 68 (42.77%) of the horse sera examined by SFDT. When T. gondii seropositivity was evaluated according to gender, it was found to be 47.92% in females and 32.92% in males. No statistical difference was observed between the gender groups (P&gt; 0.05). When T. gondii seropositivity was evaluated according to age, seropositivity rate in the equidae older than 10 years was found to be higher as 46.67%, but no statistical significance was observed among the age groups. T. gondii antibody was detected in 47 (92.16%) of donkey sera examined by SFDT. When T. gondii seropositivity was evaluated according to gender, the rate of seropositivity was found to be 89.47% in females and 93.75% in males. No statistical significance was observed between the gender groups (P&gt;0.05). When T. gondii seropositivity was evaluated according to age, the seropositivity rate in those older than 10 years was found to be higher as 96.30%, but no statistical significance was observed among the age groups (P&gt;0.05). As a result of scanning the equidea in Muş province by SFDT, seropositivity rate was found as 42.77% in horses and 92.16% in donkeys.Keywords: Donkey, Horse, SFDT, Toxoplasma gondii, Toxoplasmosis

Determination and pharmacokinetics study of UK-5099 in mouse plasma by LC\u2013MS/MS

BackgroundUK-5099 is a potent mitochondrial acetone carrier inhibitor, that exhibits anticancer activity. Recently, the anti-Toxoplasma gondii activity of UK-5099 was proposed, and in vivo studies of its pharmacokinetics in BALB/c mice are necessary to further evaluate the clinical effect of UK-5099.Methods and resultsA simple and fast high-performance liquid chromatography-tandem mass spectrometry (HPLC–MS/MS) analysis method was established and verified in terms of its linearity, matrix effect, accuracy, precision, recovery and stability. The analytes were separated by an Agilent ZORBAX XDB-C18 column (2.1 × 50 mm, 3.5 μm) at 30 °C. A gradient mobile phase consisting of water with 0.1% formic acid (FA) (phase A) and acetonitrile (ACN) (phase B) was delivered at a flow rate of 0.40 mL·min−1 with an injection volume of 5 μL. A good linear response was obtained in a concentration range of 5–5000 ng·mL−1 (r2 = 0.9947). The lower limit of quantification (LLOQ) was 5 ng·mL−1. The extraction recovery of UK-5099 was greater than 95%. The inter- and intra-day accuracy and precision of the method showed relative standard deviations (RSDs) of less than 15%. This method has been successfully applied to the pharmacokinetic evaluation of UK-5099 in mouse plasma. In health mice, the main pharmacokinetic parameters of UK-5099 after intraperitoneal administration were measured using a noncompartmental model, in which the AUC0-t was 42,103 ± 12,072 ng·h·mL−1 and the MRT0-t was 0.857 ± 0.143 h. The peak concentration (Cmax) was 82,500 ± 20,745 ng·h·mL−1, which occurred at a peak time (Tmax) = 0.250 ± 0.000 h.ConclusionsA fast and sensitive HPLC–MS/MS method was developed, validated and successfully used for the determination of UK-5099 levels in mice after intraperitoneal administration. This study was the first report of the pharmacokinetic parameters of UK-5099 in mice, which will help to further study the administration of UK-5099 in animals and humans.

Detection of Antibodies Against Toxoplasma gondii in Filter Paper-Dried Blood Dot Spots Compared with Serum in Pigs and Assessment of Variation Associated with Packed Cell Volume.

The aim of this study was to assess the agreement between anti-Toxoplasma gondii IgG antibody detection in serum and filter paper (FP) blood spots using the indirect immunofluorescence antibody assay (IFA) and to evaluate the potential impact of the packed cell volume (PCV) on antibody detection in FPs. A pair of a serum and an FP sample was collected from 96 sows at various farms in Greece, with previously identified high seropositivity and/or risk factors associated with high seropositivity against T. gondii. The PCV value was determined using the microhematocrit method. IFA was used for the detection of antibodies against T. gondii. T. gondii-specific antibodies were detected in 45.8% serum samples and 41.6% FP samples showing almost perfect agreement. Detection in FP samples presented high sensitivity (87.1-92.8%) and excellent specificity (100%) when compared with detection in serum, regardless of the PCV values. The findings of this study support the reliability of FPs for the evaluation of the serological status of swine against T. gondii. FPs could be a good alternative sample type compared with serum for large-scale epidemiological studies.

Exploring the Antiparasitic Activity of Tris-1,3,4-Thiadiazoles against Toxoplasma gondii-Infected Mice.

Nitrogen-containing atoms in their core structures have been exclusive building blocks in drug discovery and development. One of the most significant and well-known heterocycles is the 1,3,4-thidiazole nucleus, which is found in a wide range of natural products and therapeutic agents. In the present work, certain tris-1,3,4-thiadiazole derivatives (6, 7) were synthesized through a multi-step synthesis approach. All synthesized compounds were characterized using different spectroscopic tools. Previously, thiadiazole compounds as anti-Toxoplasma gondii agents have been conducted and reported in vitro. However, this is the first study to test the anti-Toxoplasma gondii activity of manufactured molecular hybrids thiadiazole in an infected mouse model with the acute RH strain of T. gondii. All the observed results demonstrated compound (7)’s powerful activity, with a considerable reduction in the parasite count reaching 82.6% in brain tissues, followed by liver and spleen tissues (65.35 and 64.81%, respectively). Inflammatory and anti-inflammatory cytokines assessments proved that Compound 7 possesses potent antiparasitic effect. Furthermore, docking tests against TgCDPK1 and ROP18 kinase (two major enzymes involved in parasite invasion and egression) demonstrated compound 7’s higher potency compared to compound 6 and megazol. According to the mentioned results, tris-1,3,4-thiadiazole derivatives under test can be employed as potent antiparasitic agents against the acute RH strain of T. gondii.