Treatment Of Toxoplasmosis Research Articles

Pancytopenia as complication after toxoplasmosis treatment – A case report

Pancytopenia as complication after toxoplasmosis treatment – A case report

Systematic Identification of Thiosemicarbazides for Inhibition of Toxoplasma gondii Growth In Vitro.

One of the key stages in the development of new therapies in the treatment of toxoplasmosis is the identification of new non-toxic small molecules with high specificity to Toxoplasma gondii. In the search for such structures, thiosemicarbazide-based compounds have emerged as a novel and promising leads. Here, a series of imidazole-thiosemicarbazides with suitable properties for CNS penetration was evaluated to determine the structural requirements needed for potent anti-Toxoplasma gondii activity. The best 4-arylthiosemicarbazides 3 and 4 showed much higher potency when compared to sulfadiazine at concentrations that are non-toxic to the host cells, indicating a high selectivity of their anti-toxoplasma activity.

Gestational and congenital toxoplasmosis - the clinicial findings in a teaching hospital

Purpose: Toxoplasmosis is a cosmopolitan zoonosis caused by the intracellular parasite Toxoplasma gondii, usually asymptomatic, in pregnant women may lead to fetuses threat of miscarriage or disabilities. There is no official coutrywide protocol describing laboratory and clinical protocols to be used in prenatal and postnatal care. Here we describe the clinical findings in pregnant women's with clinical suspicion of gestacional toxoplasmosis and as well as in their newborns until their first 18 months of life. Methods & Materials: A retrospective study was conducted and 49 medical records of pregnant women who received prenatal monitoring at the High-Risk Pregnancy Clinic of the Base Hospital of Regional School of Medicine Foundation (FUNFARME) in 2009 to 2013 were evaluated according to: gestational age, recommended treatment, obstetrical ultrasounds, clinical and laboratory diagnosis, which included IgM and IgG serologies and amniotic fluid PCRs. 39 records of their potencially infected newborns were screened observing: neurological, visual and otologic development and exams, prematurity and recommended treatment. Results: The average age of the 49 pregnant women was 23,6 ± 6,3 (min: 13; max: 39; median: 23); 22,4% (n = 11) were primigravidae and 42,8% (n = 21) multigravidae.; 75,5% (n = 37) of the pregnants presented positive serology; 46,9% (n = 23) underwent amniocentesis, 20,4% (n = 10) had a positive amniotic fluid PCR and 8,16% (n = 4) fetal ultrasound scans showed changes (shortened long bones, retrocorionic hematoma, retroamniotic hematoma and hyperechogenic intracardiac focus). Their recommended treatment included spiramycin or the triple scheme (sulfadiazine, pyrimethamine and folinic acid). Among the babies were presented: positive IgM serology (2,5% with n = 1), or positive blood PCR (7,69% with n = 3) or suspicion and signs of clinical changes (17,94% with n = 7) as cerebral calcifications, schizencephaly, seizures, chorioretinal alterations and prematurity. Although only 17,94% (n = 7) of the children underwent treatment for congenital toxoplasmosis, based on the triple scheme, not necessarily those with clinical features. Conclusion: The gestational toxoplasmosis is occurring in multigravidae more than in primigravidae. The amount of positive amniotic fluid PCRs confirm a reasonable number of fetal infection, confirming congenital toxoplasmosis; however, the number of children underwent the treatment was very low.

Anti-parasitic effect on Toxoplasma gondii induced by a spider peptide lycosin-I

Toxoplasmosis is a widely distributed parasitic protozoan disease, caused by Toxoplasma gondii (T. gondii). High prevalence of toxoplasmosis and limitations of conventional treatments lead to a search for new therapeutic drugs. Lycosin-I is a linear peptide, derived from the venom of the spider Lycosa singoriensis. The aim of the present study was to determine the anti-parasitic effect of lycosin-Ι against T. gondii. In vitro, the anti-T. gondii activities of lycosin-Ι were evaluated by MTT assay, trypan blue exclusion assay, cell counting assay and plaque assay. Cytokines of IL-6 and IL-8 were measured by quantitative PCR. In addition, the structures of tachyzoites treated with lycosin-Ι were also observed by scanning and transmission electron microscopy. In vivo, mice were challenged with parasites treated by lycosin-I. The results revealed that lycosin-Ι had shown a significant ability to inhibit T. gondii invasion and proliferation. Cytokines of IL-6 and IL-8 were reduced by lycosin-Ι at transcription level in human foreskin fibroblast (HFF) cells infected with T. gondii tachyzoites, but they were increased compared to non-infected cells. For tachyzoites, lycosin-Ι induced their cell membrane alterations with formation of invaginations, some of them appeared to be vacuolated in their cytoplasm. Moreover, lycosin-Ι had prolonged the survival time of mice by controlling T. gondii proliferation. In conclusion, our present study provides the first evidence for anti-T. gondii by using the spider peptide lycosin-Ι. These findings suggest that lycosin-Ι is a potential alternative agent for the treatment of toxoplasmosis.

Discovery of Selective Toxoplasma gondii Dihydrofolate Reductase Inhibitors for the Treatment of Toxoplasmosis.

A safer treatment for toxoplasmosis would be achieved by improving the selectivity and potency of dihydrofolate reductase (DHFR) inhibitors, such as pyrimethamine (1), for Toxoplasma gondii DHFR ( TgDHFR) relative to human DHFR ( hDHFR). We previously reported on the identification of meta-biphenyl analog 2, designed by in silico modeling of key differences in the binding pocket between TgDHFR and hDHFR. Compound 2 improves TgDHFR selectivity 6.6-fold and potency 16-fold relative to 1. Here, we report on the optimization and structure-activity relationships of this arylpiperazine series leading to the discovery of 5-(4-(3-(2-methoxypyrimidin-5-yl)phenyl)piperazin-1-yl)pyrimidine-2,4-diamine 3. Compound 3 has a TgDHFR IC50 of 1.57 ± 0.11 nM and a hDHFR to TgDHFR selectivity ratio of 196, making it 89-fold more potent and 16-fold more selective than 1. Compound 3 was highly effective in control of acute infection by highly virulent strains of T. gondii in the murine model, and it possesses the best combination of selectivity, potency, and prerequisite drug-like properties to advance into IND-enabling, preclinical development.

Clindamycin as single therapy for cerebral toxoplasmosis HIV in the history of sulfa allergy: A Case Report

Cerebral toxoplasmosis is an opportunistic infection in HIV with CD4 T-cells below < 200/uL. The aetiology is Toxoplasma gondii. Trimethoprim plus sulfamethoxazole (cotrimoxazole) combined pyrimethamine is the first line for the treatment of cerebral toxoplasmosis in immunodeficiency. The second line for treatment is clindamycin combined pyrimethamine in Indonesia. We reported a 48-year-old woman as diagnosed retroviral infection referred to Wangaya General Hospital, with main complaints of vomiting and severe headache. The blood test showed normal limits except lymphocyte account. IgM antibody for toxoplasmosis raised. Brain computed tomography-scan showed a cystic lesion in right basal ganglia with ring-enhancing surrounding by finger-like oedema. She treated with pyrimethamine and cotrimoxazole. The patient had a reddish rash on the skin with thrombocytopenia during 7-day on treatment suspected sulfa and pyrimethamine adverse side effect. Patient treated clindamycin as single therapy for cerebral toxoplasmosis. Her headache is better within two weeks of admission. Thirty days later the patient was repeated brain-CT scan, and the mass was found to be smaller. Conclusions: Single-therapy with clindamycin in CNS toxoplasmosis showed good outcome in resource-poor setting and history of sulfonamide allergy.

HIV-Related Cerebral Toxoplasmosis Revisited: Current Concepts and Controversies of an Old Disease.

Cerebral toxoplasmosis is the most common cause of expansive brain lesions in people living with HIV/AIDS (PLWHA) and continues to cause high morbidity and mortality. The most frequent characteristics are focal subacute neurological deficits and ring-enhancing brain lesions in the basal ganglia, but the spectrum of clinical and neuroradiological manifestations is broad. Early initiation of antitoxoplasma therapy is an important feature of the diagnostic approach of expansive brain lesions in PLWHA. Pyrimethamine-based regimens and trimethoprim-sulfamethoxazole (TMP-SMX) seem to present similar efficacy, but TMP-SMX shows potential practical advantages. The immune reconstitution inflammatory syndrome is uncommon in cerebral toxoplasmosis, and we now have more effective, safe, and friendly combined antiretroviral therapy (cART) options. As a consequence of these 2 variables, the initiation of cART can be performed within 2 weeks after initiation of antitoxoplasma therapy. Herein, we will review historical and current concepts of epidemiology, diagnosis, and treatment of HIV-related cerebral toxoplasmosis.

Endogenous Endophthalmitis by &amp;lt;i&amp;gt;Candida Albicans&amp;lt;/i&amp;gt; Associated with a Primary Immunodeficiency Due to CARD9 Deficiency

This report encopasses a case of endogenous endophthalmitis by <i>Candida albicans</i> in an otherwise healthy young adult patient, which led to the diagnosis of a primary immunodeficiency, CARD9 deficiency, associated with familial candidiasis. A 31-year-old female patient with a one-month history of low visual acuity (VA) in the right eye (RE) was referred after no response to treatment of toxoplasmosis and acute retinal necrosis. She presented no comorbidities, but a history of recurrent paronychia, oral candidiasis and <i>tinea capitis</i> since childhood. Her parents were first-degree cousins. Visual acuity was 20/2000 in the RE and 20/20 in the left eye (LE). There was an elevated exudative lesion occupying the retinal posterior pole, associated with fluffy white cotton balls in the adjacent posterior vitreous. Optical coherence tomography (OCT) showed macular lesions with the "rain cloud" sign. Fungal endogenous endophthalmitis was hypothesized. Imaging and laboratory examinations discarded other infections and eliminated involvement of other organs. Diagnostic vitreous puncture was performed in addition to Amphotericin B injection and intravenous Fluconazole. As culture results were negative, a vitreous biopsy was performed, in which <i>Candida albicans</i> was grown, followed by a second Amphotericin B injection. As there was no improvement, pars plana posterior vitrectomy, with placement of silicone oil, was performed. In spite of infection control, VA remained 20/2000. Due to a severe ocular condition with no other invasive sites of infection, associated with superficial recurrent fungal infections, a primary fungal specific immunodeficiency was suspected, resulting in referral to Immunology. An immunogenetic panel was performed, demonstrating heterozygosity for two CARD9 gene mutations, a molecule related to familial candidiasis. Follow-up in Ophthalmology and Immunology was maintained and oral Fluconazole was proposed to be taken for at least 6 months. Three months later, she presented with an oral abscess secondary to <i>Candida</i>, managed by the Otorhinolaryngology team. Fungal endogenous endophthalmitis is an important cause of morbidity and low visual acuity, especially in the population of immunocompromised patients. Early ophthalmological diagnosis should preferably be performed in order to prevent lesions from increasing, affecting noble areas such as the macula, and reaching the vitreous. When the fundus exam presents signs of fungal endophthalmitis in immunocompetent, young and healthy patients, diagnosis of primary immunodeficiencies should be considered.

1895. The Anti-Parasitic Drug Atovaquone Inhibits Arbovirus Replication

BackgroundAtovaquone, a hydroxynaphthoquinone, FDA Pregnancy category C, used for the treatment and prevention of pneumocysits jirovecii pneumonia (PCP), toxoplasmosis, babesiosis and malaria has in vitro activity against Zika virus (ZIKV). The mechanism of action against Plasmodium spp. and other parasites is based in the inhibition of mitochondrial cytochrome bc1 complex which further collapses parasite–mitochondrial membrane potential. But to date, antiviral activity of this drug has not been described.MethodsVero cells (monkey kidney epithelial cells) were seeded. At 24 hours of incubation, the cells were pretreated with ribavirin and brequinar (known antiviral drugs) and atovaquone at different concentrations for 1 hour and then infected with ZIKV Brazilian strain and Ugandan strain, and subsequently treated with drugs again. After incubation for 72 hours virus antigen Env-protein production was quantified by immunodetection. The concentration of atovaquone that decreased the level of Env-protein production by 50% was calculated by non-linear regression analysis (CC50). Cell viability was measured using the CellTiter 96 aqueous one solution cell proliferation assay (Promega, Madison, WI), according to the manufacturers protocol. Viral infection was rescued adding uracil to Vero cells pre-treated with ribavirin, brequinar and atovaquone. Experiment was repeated with Chikungunya virus (CHIKV).ResultsWe found that atovaquone inhibits ZIKV infection in Vero cells at smaller concentration (CC50 = 0.52 μM) than those used for parasitic killing. The effect is more prominent in the Brazilian strain when compared with the Ugandan strain. No cytotoxic effect was found in Vero cells up to 15 μM; above this concentration atovaquone formed crystals. Uracil rescues ZIKV infection after treatment with atovaquone. Atovaquone also inhibited CHIKV infection in Vero cells. ConclusionAtovaquone has antiviral activity against ZIKV likely via depletion of nucleotides blocking pyrimidine biosynthesis. Furthermore, the antiviral effect is applicable to other arboviruses which makes atovaquone a broad-spectrum antiviral drug and a potential attractive candidate for the treatment of ZIKV infection in vulnerable population such pregnant women and children.Disclosures All authors: No reported disclosures.

USE OF MIDDLE-FIELD MRIWITH SPECIFIC SEQUENCIES COMPENSATING THE MOVEMENT ARTEFACTS FOR BRAIN STUDIES IN PATIENTS WITH HIV INFECTION

Nowadays of great importance is not only the issue of early diagnosis of HIV infection, but of early detection and effective treatment of AIDS complications. The annual increase in the incidence of HIV infection amounts to 4%. Unfortunately, a great number of patients ask for hospital care when their disease has the stage of secondary complications. Such situation requires a fast and affordable diagnostics together with verification of the nature of the affection. Purpose. To evaluate the diagnosis effectiveness of the secondary infectious cerebral affection in patients with HIV infection using the method of non-contrast MRI including the usage of the specially developed software. One hundred thirty three patients were recruited for the study, the main group of 108 persons (as old as 36,8±8,3), all referred to the brain MRI with suggestion of neurologic complications of HIV. The control group comprised 25 patients of the same age with focal damage of vascular nature. In everybody the MRI has been carried out using open middle-field scanner (Az-360, by AZ plc company, Moscow) with field induction 0,4 T, supplied with wireless four-channel quadratur coil for head studies, without contrast enhancement, but using in everybody the specially designed protocols for compensation of movement artifacts. All patients were re-examined in 2, 4 and 6 weeks. MRI images both in first admission and in follow-up studies were reported first qualitatively by type of contour of pathologic focus, by presence of multiple pattern of focal damage, by ultrastructure of foci, extent of perifocal oedema, interhaemispheric dislocation; also the dimensions of lateral ventriculi, of external subarachnoidal spaces and thickness of cortex in various regions were measured. Focal cerebral damage verified later as toxoplasmosis was revealed in the main group in 80% (86 of 108) of patients. False-negative and false-positive conclusions of MRI studies were not revealed also in prospective follow-up studies. Of these in 69 (80,2%) the pathologic foci were multiple and did localize periventricularly in particular in putamen, nucleus caudatus, equally frequent for both haemispherae, single foci were detected in thalami, concomitant with tendency to extensive cortical dystrophy. The putamen region was involved more frequently as compared to other basal ganglii (р=0,003). No significant differences were revealed between various cortical regions as regard to frequency of detection of pathologic foci. Four weeks period was detected as being the most effective time for the dynamic control in the course of treatment. Conclusion. Middle-field MRI of the brain with compensation of movement artifacts is a proper technique for both the diagnosis and follow-up control of treatment of cerebral toxoplasmosis in patients with HIV infection.

Curcumin nanoemulsion as a novel chemical for the treatment of acute and chronic toxoplasmosis in mice.

BackgroundThe aim of this study was to prepare curcumin nanoemulsion (CR-NE) to solve the problems associated with poor water solubility and low bioavailability of CR and to test its efficiency in the treatment of acute and chronic toxoplasmosis in mouse models.Materials and methodsCR-NE 1% was prepared using spontaneous emulsification by soybean as oil phase; a mixture of Tween 80 and Tween 85 as surfactant; ethanol as cosurfactant and distilled water. Particle size and zeta potential of NE were assessed using Nano-ZS90 dynamic light scattering. Stability testing of NE was assessed after storage for 2 months at room temperature. In vivo experiments were carried out using 50 BALB/c mice inoculated with virulent RH strain (type I) and 50 BALB/c mice inoculated with avirulent Tehran strain (type II) of Toxoplasma gondii and treated with CR-NE (1% w/v), CR suspension (CR-S, 1% w/v), and NE without CR (NE-no CR).ResultsThe mean particle size and zeta potential of CR-NE included 215.66±16.8 nm and −29.46±2.65 mV, respectively, and were stable in particle size after a three freeze–thaw cycle. In acute phase experiment, the survival time of mice infected with RH strain of T. gondii and treated with CR-NE extended from 8 to 10 days postinoculation. The differences were statistically significant between the survival time of mice in CR-NE-treated group compared with negative control group (P<0.001). Furthermore, CR-NE significantly decreased the mean counts of peritoneum tachyzoites from 5,962.5±666 in negative control group to 627.5±73 in CR-NE-treated mice (P<0.001). Growth inhibition rates of tachyzoites in peritoneum of mice receiving CR-NE, CR-S, and NE-no CR included 90%, 21%, and 11%, respectively, compared with negative control group. In chronic phase experiment, the average number and size of tissue cysts significantly decreased to 17.2±15.6 and 31.5±6.26 µm, respectively, in mice inoculated with bradyzoites of T. gondii Tehran strain and treated with CR-NE compared with that in negative control group (P<0.001). Decrease of cyst numbers was verified by downregulation of BAG1 in treatment groups compared with negative control group with a minimum relative expression in CR-NE (1.12±0.28), CR-S (11.76±0.87), and NE-no CR (14.67±0.77), respectively, (P<0.001).ConclusionResults from the current study showed the potential of CR-S and CR-NE in treatment of acute and chronic toxoplasmosis in mouse models for the first time. However, CR-NE was more efficient than CR-S, and it seems that CR-NE has a potential formula for the treatment of acute and chronic toxoplasmosis, especially in those with latent bradyzoites in brain.

Drug Resistance in Toxoplasma gondii.

Toxoplasma gondii (T. gondii) is a global protozoan parasite infecting up to one-third of the world population. Pyrimethamine (PYR) and sulfadiazine (SDZ) are the most widely used drugs for treatment of toxoplasmosis; however, several failure cases have been recorded as well; suggesting the existence of drug resistant strains. This review aims to give a systematic and comprehensive understanding of drug resistance in T. gondii including mechanisms of resistance and sites of drug action in parasite. Analogous amino acid substitutions in the Toxoplasma enzyme were identified to confer PYR resistance. Moreover, resistance to clindamycin, spiramycin, and azithromycin is encoded in the rRNA genes of T. gondii. However, T. gondii SDZ resistance mechanism has not been proved yet. Recently there has been a slight increase in SDZ resistance. That is why the majority of studies were carried out using SDZ. Six strains resistant to SDZ were found in clinical cases between 2013 and 2017 which among Brazilian T. gondii isolates, TgCTBr11, Ck3, and Pg1 were identified in human toxoplasmosis, as well as in livestock intended for human consumption. In conclusion, recent experimental studies in clinical cases have clearly shown that drug resistance in Toxoplasma is ongoing. Thus, establishing a more effective therapeutic scheme in the treatment of toxoplasmosis is critically needed. The emergence of T. gondii strains resistant to current drugs, reviewed here, represents a concern not only for treatment failure but also for increased clinical severity in immunocompromised patients. To improve the therapeutic outcome in patients, a greater understanding of the exact mechanisms of drug resistance in T. gondii should be developed. Thus, monitoring the presence of resistant parasites, in food products, would seem a prudent public health program.

Could Araucaria heterophylla resin extract be used as a new treatment for toxoplasmosis?

Toxoplasmosis is a worldwide parasitic disease responsible for serious health problems to human. The currently available drugs used for toxoplasmosis treatment showed a limited efficacy and cause serious host toxicity. The in vitro screening for toxoplasmicidal activity of Araucaria heterophylla resin (AHR) extract and its major component 13-epi-cupressic acid (CUP) showed that both AHR (EC50 = 3.90) and CUP (EC50 = 3.69) have high toxoplasmicidal activity in comparison with standard cotrimoxazole (EC50 = 4.28). The antiprotozoal effects of AHR and CUP were investigated against acute and chronic toxoplasmosis using mice models. Two groups of Swiss albino mice were infected by RH Toxoplasma strain intraperitoneally and by Me49 strain orally. Both groups were treated with AHR and CUP in different doses. Their effects were evaluated by survival rate, peritoneal, spleen and liver parasite burdens, brain cyst burden, NO serum level and histopathological lesions. The ultrastructural changes of tachyzoites of acutely infected mice were studied using scanning electron microscopy (SEM). There is an evidence of toxoplasmicidal activity of AHR and CUP in acute and chronic experimental toxoplasmosis. In the acute model, mice treated with AHR and CUP showed prolonged survival rates, a significant decrease in the parasite density in peritoneal lavage and pathological insult in both liver and spleen compared with that of untreated ones. SEM results denote evident morphological alterations of treated tachyzoites. In chronic experimental toxoplasmosis, AHR and CUP treated groups could significantly reduce brain cyst burden by 96.05% and 98.02% respectively. This study indicates that AHR and CUP showed potent toxoplasmicidal activities experimentally and could be used as a potential natural nontoxic agent for treatment of toxoplasmosis.

Treatment of Toxoplasmosis: Historical Perspective, Animal Models, and Current Clinical Practice.

Primary Toxoplasma gondii infection is usually subclinical, but cervical lymphadenopathy or ocular disease can be present in some patients. Active infection is characterized by tachyzoites, while tissue cysts characterize latent disease. Infection in the fetus and in immunocompromised patients can cause devastating disease. The combination of pyrimethamine and sulfadiazine (pyr-sulf), targeting the active stage of the infection, is the current gold standard for treating toxoplasmosis, but failure rates remain significant. Although other regimens are available, including pyrimethamine in combination with clindamycin, atovaquone, clarithromycin, or azithromycin or monotherapy with trimethoprim-sulfamethoxazole (TMP-SMX) or atovaquone, none have been found to be superior to pyr-sulf, and no regimen is active against the latent stage of the infection. Furthermore, the efficacy of these regimens against ocular disease remains uncertain. In multiple studies, systematic screening for Toxoplasma infection during gestation, followed by treatment with spiramycin for acute maternal infections and with pyr-sulf for those with established fetal infection, has been shown to be effective at preventing vertical transmission and minimizing the severity of congenital toxoplasmosis (CT). Despite significant progress in treating human disease, there is a strong impetus to develop novel therapeutics for both the acute and latent forms of the infection. Here we present an overview of toxoplasmosis treatment in humans and in animal models. Additional research is needed to identify novel drugs by use of innovative high-throughput screening technologies and to improve experimental models to reflect human disease. Such advances will pave the way for lead candidates to be tested in thoroughly designed clinical trials in defined patient populations.

Gestational and congenital toxoplasmosis: Report of a clinical evaluation in Brazil

Background: Toxoplasmosis is a cosmopolitan zoonosis caused by the intracellular parasite Toxoplasma gondii. Despite usually asymptomatic, in pregnant women, may lead to fetuses threat of miscarriage or disabilities. To diagnose gestational and congenital/fetal infection, must be analyzed the IgM and IgG serologies as well the PCR in amniotic fluid. The main purpose was to describe the clinical aspects from pregnant women's with clinical suspicion of gestacional toxoplasmosis as well from their newborns until their first 18 months of life. Methods & Materials: A retrospective study was conducted in a tertiary teaching hospital in São Paulo state, Brazil and 49 medical records of pregnant women who received prenatal monitoring at the High-Risk Pregnancy Clinic of the Hospital de Base from the Regional School of Medicine Foundation (FUNFARME) in 2009 to 2013 were evaluated according to: gestational age, recommended treatment, obstetrical ultrasounds, clinical and laboratory diagnosis, which included IgM and IgG serologies and amniotic fluid PCRs. 39 records of their potencially infected newborns were screened observing: neurological, visual and otologic development and exams, prematurity and recommended treatment. Results: The average age of the 49 pregnant women was 23,6 ± 6,3 (min: 13; max: 39; median: 23); 75,5% (n = 37) presented positive serology for toxoplasmosis; 46,9% (n = 23) underwent amniocentesis, 20,4% (n = 10) had a positive amniotic fluid PCR and 8,16% (n = 4) fetal ultrasound scans showed changes (shortened long bones, retrocorionic hematoma, retroamniotic hematoma and hyperechogenic intracardiac focus). Their recommended treatment included spiramycin or the triple scheme (sulfadiazine, pyrimethamine and folinic acid). Among the babies were presented: positive IgM serology (2,5% with n = 1), or positive blood PCR (7,69% with n = 3) or suspicion and signs of clinical changes (17,94% with n = 7) as cerebral calcifications, schizencephaly, seizures, chorioretinal alterations and prematurity. Although only 17,94% (n = 7) underwent treatment for congenital toxoplasmosis, based on the triple scheme, not necessarily those with clinical features. Conclusion: The toxoplasmosis prevalience of gestational infection is high indicating the epidemiological relevance of toxoplasmosis in this region. The positive PCR in amniotic fluid confirm a reasonable number of fetal infection; However, the number of children underwent the recommended treatment was low.

Seroprevalence and preliminary treatment of toxoplasmosis of pregnant goats in Kalubyia Gobernatore, Egypt

Toxoplasmosis is a zoonotic disease that commonly infects goat in Egypt. For its diagnosis, sera form 48 pregnant does were tested for Toxoplasma gondii antibodies through indirect haemagglutination test (IHAT) and modified agglutination test (MAT). Samples were collected between the periods extended from November 2006 to March 2007. The seropositive rate of pregnant does (Meet Kenana, Kalubyia Governorate, Egypt) was 35.42% (17/48). After using MAT test against T. gondii- IgM, 11 sera (22.9%) were found positive. The older animals were more frequently infected (50%) than the younger goats (26.3%). Higher prevalence in aged does was confirmed by MAT. Total T.gondii antibodies and IgM were gradually increased with the development of pregnancy. Two months post treatment with sulfademidine 33.3 %, for 5 successive days at a dose of 200 mg/kg, resulted in not only reduction of T.gondii total antibodies and IgM in all treated does, but re-modulation of T.gondii antibody titres moderately toward the mild titres, and prevention of congenital infection and abortion as well. Whereas IgM was not recorded in treated does, it was detected in the untreated ones. No abortion was recorded in all treated does, but an untreated doe delivered two dead fetuses. The delivered kids belonged to treated does were seropositive for T. gondii antibodies by using IHAT, yet they were seronegative for T.gondii‐ IgM by using MAT. In contrast, kids belonged to untreated does were seropositive by using both IHAT and MAT. Adequate management might be useful and essential to control toxoplasmosis in the goat herds of Kalubyia Governorate.

Transcriptomic analysis reveals Toxoplasma gondii strain-specific differences in host cell response to dense granule protein GRA15.

Growth and replication of the protozoan parasite Toxoplasma gondii within host cell entail the production of several effector proteins, which the parasite exploits for counteracting the host's immune response. Despite considerable research to define the host signaling pathways manipulated by T. gondii and their effectors, there has been limited progress into understanding how individual members of the dense granule proteins (GRAs) modulate gene expression within host cells. The aim of this study was to evaluate whether T. gondii GRA15 protein plays any role in regulating host gene expression. Baby hamster kidney cells (BHK-21) were transfected with plasmids encoding GRA15 genes of either type I GT1 strain (GRA15I) or type II PRU strain (GRA15II). Gene expression patterns of transfected and nontransfected BHK-21 cells were investigated using RNA-sequencing analysis. GRA15I and GRA15II induced both known and novel transcriptional changes in the transfected BHK-21 cells compared with nontransfected cells. Pathway analysis revealed that GRA15II was mainly involved in the regulation of tumor necrosis factor (TNF), NF-κB, HTLV-I infection, and NOD-like receptor signaling pathways. GRA15I preferentially influenced the synthesis of unsaturated fatty acids in host cells. Our findings support the hypothesis that certain functions of GRA15 protein are strain dependent and that GRA15 modulates the expression of signaling pathways and genes with important roles in T. gondii pathophysiology. A greater understanding of host signaling pathways influenced by T. gondii effectors would allow the development of more efficient anti-T. gondii therapeutic schemes, capitalizing on disrupting parasite virulence factors to advance the treatment of toxoplasmosis.

Treatment of Toxoplasmosis and Neosporosis in Farm Ruminants: State of Knowledge and Future Trends.

Toxoplasmosis and neosporosis are closely related protozoan diseases that lead to important economic impacts in farm ruminants. Toxoplasma gondii infection mainly causes reproductive failure in small ruminants and is a widespread zoonosis, whereas Neospora caninum infection is one of the most important causes of abortion in cattle worldwide. Vaccination has been considered the most economic measure for controlling these diseases. However, despite vaccine development efforts, only a live-attenuated T. gondii vaccine has been licensed for veterinary use, and no promising vaccines against ne-osporosis have been developed; therefore, vaccine development remains a key goal. Additionally, drug therapy could be a valuable strategy for disease control in farm ruminants, as several drugs that limit T. gondii and N. caninum proliferation and dissemination have been evaluated. This approach may also be relevant to performing an initial drug screening for potential human therapy for zoonotic parasites. Treat-ments can be applied against infections in adult ruminants to minimize the outcomes of a primo-infection or the reactivation of a chronic infection during gestation or in newborn ruminants to avoid infection chronification. In this review, the current status of drug development against toxoplasmosis and neosporo-sis in farm ruminants is presented, and in an effort to promote additional treatment options, prospective drugs that have shown efficacy in vitro and in laboratory animal models of toxoplasmosis and neosporosis are examined

Structural predication and antigenic analysis of Toxoplasma gondii ROP20.

Toxoplasma gondii infects almost all the warm-blooded animals. ROP20 protein is expressed in the rhoptry of Toxoplasma gondii. In this study, the secondary structure of ROP20 was analyzed using SMART software. We constructed and analyzed the 3D model of ROP20 protein using SWISS-MODEL online procedure and Visual Molecular Dynamics (VMD) software. The structure analysis fully indicated that ROP20 protein is an important member of the ROP family. Furthermore, We used DNASTAR software and Epitope Database online service to analyze liner-B cell epitopes and T-cell epitopes of ROP20 protein. All the analysis results of ROP20 protein can provide positive information on treatment and vaccine for toxoplasmosis. Moreover, ROP20 gene was obtained from PCR, and a recombinant eukaryotic expression vector (pEGFP-C1-ROP20) was constructed in the following study. After restriction enzyme digestion, the constructed plasmid was transfected into HEK 293-T cells. The RT-PCR result indicated that the recombinant plasmid could transcribe successfully in HEK 293-T cell. The results of western blotting indicated the expressed proteins can be recognized by anti-STAg mouse sera.

Ocular toxoplasmosis: adverse reactions to treatment in a Brazilian cohort.

The purpose of this study was to estimate the frequency and describe the adverse drug reactions (ADRs) associated with the classic treatment of ocular toxoplasmosis (OT), namely sulfadiazine, pyrimethamine, corticosteroids and folinic acid. We performed a descriptive study of a prospective cohort of patients with OT treated with the classic therapy. Data were collected during medical consultations and treatment. Of the 147 patients studied, 85% developed one or more ADR. Women presented more ADRs than men (95% vs 77%). Of the total reactions (n=394), 82% were mild, but we found one life-threatening event (Stevens-Johnson syndrome). The most frequent types (71%) of ADRs were gastrointestinal, skin and neurological or psychiatric. The majority of ADRs (90.3%) occurred before the second week of treatment. A third of the patients were treated for the ADR and 10% dropped out of OT treatment. Most (70%) of the ADRs were characterized as being probably caused by the drugs and may be associated with prednisone, sulfadiazine and sulfadiazine/prednisone. Six percent of ADRs were not previously described, such as taste alteration, constipation/bloating, dyspnoea, sweating and somnolence. Our results suggest a high rate of ADRs to OT classic treatment, which requires careful follow-up in order to identify and treat ADRs early.