Pregnant Mouse Model Research Articles

Teratogenic effect of the aqueous extract of the Foeniculum vulgare (fennel) on fetal development in mice

Objective The present study evaluated the effect of aqueous extract of Foeniculum vulgare on fetal development in pregnant mice models. Methods A total number of 24 female BALB/c mice with a weight range of 25-30 gr divided into four groups. Each group received 0.25 ml of the aqueous extract of Foeniculum vulgare with different concentrations (2.5 mg, 12.5 mg, 25 mg) and distilled water as a control group. The aqueous extract of Foeniculum vulgare administered via oral gavage on a daily basis from day 6 to day 15 of pregnancy. On day 16, the fetuses were analyzed in terms of morphological changes, skeletal disorders, and cellular alterations. Results The result showed the dose-dependent teratogenic effect of the aqueous extract of Foeniculum vulgare. At 12.5 mg and 25 mg concentration, the teratogenic effect was more severe. Oral gavage administration of the aqueous extract of Foeniculum vulgare increased the number of dead fetuses and reduced the average weight, height, and crown-rump length. Subcutaneous hemorrhage, dorsal lesion, wrinkled skin, and considerably lower than normal fetal weight observed in gross morphological inspection at 25 mg concentration. The skeletal studies revealed fetal anomalies, reduction in ossification and reduction in the number of ribs. Internal bleeding around the liver and lungs, pulmonary fibrosis and disruption in the arrangement of hepatocytes was also observed in histological analysis. Conclusion Administration of the aqueous extract of Foeniculum vulgare results in embryotoxicity in mice models in morphological, skeletal and cellular levels. Keywords Foeniculum vulgare, Fennel, Teratogenicity, Embryotoxicity, Fetal Development, Mice

Teratogenic effect of the aqueous extract of the Foeniculum vulgare (fennel) on fetal development in mice

Objective The present study evaluated the effect of aqueous extract of Foeniculum vulgare on fetal development in pregnant mice models.
 Methods A total number of 24 female BALB/c mice with a weight range of 25-30 gr divided into four groups. Each group received 0.25 ml of the aqueous extract of Foeniculum vulgare with different concentrations (2.5 mg, 12.5 mg, 25 mg) and distilled water as a control group. The aqueous extract of Foeniculum vulgare administered via oral gavage on a daily basis from day 6 to day 15 of pregnancy. On day 16, the fetuses were analyzed in terms of morphological changes, skeletal disorders, and cellular alterations.
 Results The result showed the dose-dependent teratogenic effect of the aqueous extract of Foeniculum vulgare. At 12.5 mg and 25 mg concentration, the teratogenic effect was more severe. Oral gavage administration of the aqueous extract of Foeniculum vulgare increased the number of dead fetuses and reduced the average weight, height, and crown-rump length. Subcutaneous hemorrhage, dorsal lesion, wrinkled skin, and considerably lower than normal fetal weight observed in gross morphological inspection at 25 mg concentration. The skeletal studies revealed fetal anomalies, reduction in ossification and reduction in the number of ribs. Internal bleeding around the liver and lungs, pulmonary fibrosis and disruption in the arrangement of hepatocytes was also observed in histological analysis.
 Conclusion Administration of the aqueous extract of Foeniculum vulgare results in embryotoxicity in mice models in morphological, skeletal and cellular levels.
 Keywords Foeniculum vulgare, Fennel, Teratogenicity, Embryotoxicity, Fetal Development, Mice

Repurposing an HIV Drug for Zika Virus Therapy.

Repurposing an HIV Drug for Zika Virus Therapy.

The Use of Natural Phenomenon in Obtaining Pregnant Rats and Mice as Experimental Animals with The Same Gestational Ages

Hormonal oestrus synchronization is often used to obtain the same gestational ages animal models, but it may not physiologic nor practical. This research was conducted to find out a simple method in obtaining physiologic pregnant experimental animals. We used 120 female rats and mice each, divided into 16 groups with 15 members each. They were classified according to the state of parities, pair or group mating, and synchronization of oestrous. The estrous synchronization is done by utilizing the natural phenomenon (Lee-Boot, Pheromone, and Whitten effect). All nulliparous and multiparous models then were simultaneously mated for one night in pairs (1:1) or in groups (1:5) subsequently. Pregnancy rates were noted based on the number of models delivering pups on day 19-23 post-mating. The highest pregnancy rate was noted in group of multiparous, synchronized and monogamous rats (86.7%), followed by nulliparous, synchronized and monogamous rats (73.3%) and multiparous, synchronized and monogamous mice (60%) (p<0.05). Synchronization of the estrous and monogamous mating of rats or mice had significant roles in pregnancy rates (p<0.05). To get a high number of pregnant rats or mice models, we suggest to use the multiparous, estrous synchronized and monogamous simultaneous mating within one night.

HMGB1 release from trophoblasts contributes to inflammation during Brucella melitensis infection.

Brucella melitensis infection causes acute necrotizing inflammation in pregnant animals; however, the pathophysiological mechanisms leading to placentitis are unknown. Here, we demonstrate that high-mobility group box 1 (HMGB1) acts as a mediator of placenta inflammation in B.melitensis-infected pregnant mice model. HMGB1 levels were increased in trophoblasts or placental explant during B.melitensis infection. Inhibition of HMGB1 activity with neutralising antibody significantly reduced the secretion of inflammatory cytokines in B.melitensis-infected trophoblasts or placenta, whereas administration of recombinant HMGB1 (rHMGB1) increased the inflammatory response. Mechanistically, this decreased inflammatory response results from inhibition of HMGB1 activity, which cause the suppression of both mitogen-activated protein kinases and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) activation. Moreover, neutralising antibody to HMGB1 prevented B.melitensis infection-induced activation of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase in trophoblasts. In contrast, in vitro stimulation of trophoblasts with rHMGB1 caused activation of NADPH oxidase and increased the production of ROS, which contributes to high bacterial burden within trophoblasts or placenta. In vivo, treatment with anti-HMGB1 antibody increases the number of Brucella survival within placenta in B.melitensis-infected pregnant mice but successfully reduced the severity of placentitis and abortion.

A Measles Virus-Based Vaccine Candidate Mediates Protection against Zika Virus in an Allogeneic Mouse Pregnancy Model.

The impact of the Zika virus (ZIKV) epidemic highlights the need for vaccines that reduce or prevent infection and reliably prevent teratogenic complications. The live-attenuated measles virus (MV) vaccine strains are a promising vaccine platform, since they induce robust humoral and cellular immune responses against additional antigens and have an excellent safety record. To explore its potential to protect against ZIKV, we compared a recombinant Schwarz strain MV that encodes ZIKV prM and soluble E proteins (MV-Zika-sE) with a prototypic alum-adjuvanted whole inactivated ZIKV particle vaccine. Analysis of MV-Zika-sE-infected cells confirmed antigen expression, and the virus replicated with vaccine strain characteristics. Immunized IFNAR-/--CD46Ge mice developed E protein-specific and neutralizing antibodies, and ZIKV E-specific cellular immune responses were observed by gamma interferon (IFN-γ) enzyme-linked immunospot (ELISpot) and in vitro T cell proliferation assays. To analyze protective efficacy, vaccinated female mice were challenged with ZIKV after allogeneic mating. In MV-Zika-sE-vaccinated mice, weight gain was similar to that in uninfected mice, while no plasma viremia was detectable in the majority of the animals. In contrast, infected control animals gained less weight and experienced about 100-fold higher viremia over at least 3 days. Moreover, vaccination with MV-Zika-sE reduced the ZIKV load in different organs and the placentas and prevented infection of the fetus. Consequently, no fetal growth retardation, anemia, or death due to ZIKV infection was seen in MV-Zika-sE-vaccinated dams. In contrast, the inactivated ZIKV vaccine had little to no effect in our studies. Therefore, the MV-derived ZIKV vaccine is a promising candidate for further preclinical and clinical development.IMPORTANCE Zika virus (ZIKV) is a mosquito-borne flavivirus that causes a variety of neurological complications, including congenital birth defects. Despite the urgent need, no ZIKV vaccine has yet been licensed. Recombinant vaccine strain-derived measles viruses (MV) constitute a promising vector platform to induce immunity against foreign pathogens by expressing antigens from additional transcription units while at the same time possessing a remarkable safety profile. This concept has already been validated against different pathogens, including at least 3 other flaviviruses, and our data show that vaccination with MV expressing soluble ZIKV E protein significantly diminishes infection and prevents fetal loss or damage in an allogeneic mouse pregnancy model. It can thus be regarded as a promising emergency vaccine candidate with the potential for inclusion in routine vaccination settings in areas of endemicity to prevent teratogenic effects of circulating ZIKV during pregnancy, comparable to standard rubella virus vaccination.

Zika Virus: Origins, Pathological Action, and Treatment Strategies.

The Zika virus (ZIKV) global epidemic prompted the World Health Organization to declare it a 2016 Public Health Emergency of International Concern. The overwhelming experience over the past several years teaches us that ZIKV and the associated neurological complications represent a long-term world-wide challenge to public health. Although the number of ZIKV cases in the Western Hemisphere has dropped since 2016, the need for basic research and anti-ZIKV drug development remains strong. Re-emerging viruses like ZIKV are an ever-present threat in the 21st century where fast transcontinental travel lends itself to viral epidemics. Here, we first present the origin story for ZIKV and review the rapid progress researchers have made toward understanding of the ZIKV pathology and in the design, re-purposing, and testing–particularly in vivo–drug candidates for ZIKV prophylaxis and therapy ZIKV. Quite remarkably, a short, but intensive, drug-repurposing effort has already resulted in several readily available FDA-approved drugs that are capable of effectively combating the virus in infected adult mouse models and, most importantly, in both preventing maternal-fetal transmission and severe microcephaly in newborns in pregnant mouse models.

Endochin-Like Quinolones Exhibit Promising Efficacy Against Neospora Caninum in vitro and in Experimentally Infected Pregnant Mice.

We report on the efficacy of selected endochin-like quinolones (ELQs) against N. caninum tachyzoites grown in human foreskin fibroblasts (HFF), and in a pregnant BALB/c mouse model. Fourteen ELQs were screened against transgenic N. caninum tachyzoites expressing β-galactosidase (Nc-βgal). Drugs were added concomitantly to infection and the values for 50% proliferation inhibition (IC50) were determined after 3 days. Three compounds exhibited IC50 values below 0.1 nM, 3 ELQs had IC50s between 0.1 and 1 nM, for 7 compounds values between 1 and 10 nM were noted, and one compound had an IC50 of 22.4 nM. Two compounds, namely ELQ-316 and its prodrug ELQ-334 with IC50s of 0.66 and 3.33 nM, respectively, were previously shown to display promising activities against experimental toxoplasmosis and babesiosis caused by Babesia microti in mice, and were thus further studied. They were assessed in long-term treatment assays by exposure of infected HFF to ELQs at 0.5 μM concentration, starting 3 h after infection and lasting for up to 17 days followed by release of drug pressure. Results showed that the compounds substantially delayed parasite proliferation, but did not exert parasiticidal activities. TEM of drug treated parasites detected distinct alterations within the parasite mitochondria, but not in other parasite organelles. Assessment of safety of ELQ-334 in the pregnant mouse model showed that the compound did not interfere in fertility or pregnancy outcome. In N. caninum infected pregnant mice treated with ELQ-334 at 10 mg/kg/day for 5 days, neonatal mortality (within 2 days post partum) was found in 7 of 44 pups (15.9%), but no postnatal mortality was noted, and vertical transmission was reduced by 49% compared to the placebo group, which exhibited 100% vertical transmission, neonatal mortality in 15 of 34 pups (44%), and postnatal mortality for 18 of the residual 19 pups during the 4 weeks follow-up. These findings encourage more research on the use of ELQs for therapeutic options against N. caninum infection.

Elevated microRNA-141-3p in placenta of non-diabetic macrosomia regulate trophoblast proliferation.

BackgroundSeveral studies have reported microRNAs (miRNAs) could regulate the placental development, though the role and mechanism of miRNAs in the development of non-diabetic macrosomia (NDFMS) remains unclear.MethodsTo identify the aberrantly expressed key miRNAs in placenta of NDFMS, we employed a strategy consisting of initial screening with miRNA microarray and further validation with quantitative RT-PCR assay (qRT-PCR). In vitro cellular model and a mouse pregnancy model were used to delineate the functional effects of key miRNA on proliferation, invasion, and migration.FindingsmiR-141-3p was identified as the key miRNA with expression level significantly higher in placentas of NDFMS compared with those from normal controls. Overexpressed miR-141-3p in HTR-8/SVneo cells contributed to increased cell proliferation, invasion, and migration. miR-141-3p inhibition in HTR-8/SVneo cells resulted in decreased cell proliferation and invasion. Significantly increased infant birth weight was observed in late pregnancy of C57BL/6J mice treated with miR-141-3p agomir. However, no significant difference was found in early pregnancy of C57BL/6J mice treated with miR-141-3p agomir.InterpretationmiR-141-3p could stimulate placental cell proliferation to participate in the occurrence and development of NDFMS.

Improving the clinical relevance of a mouse pregnancy model of antiretroviral toxicity; a pharmacokinetic dosing-optimization study of current HIV antiretroviral regimens

Animal models can be useful tools for the study of HIV antiretroviral (ARV) safety/toxicity in pregnancy and the mechanisms that underlie ARV-associated adverse events. The utility and translatability of animal model-based ARV safety/toxicity data is improved if ARVs are tested in clinically relevant concentrations. The objective of this work was to improve the clinical relevance of our mouse pregnancy model of ARV toxicity, by determining the doses of currently prescribed ARV regimens that would yield human therapeutic plasma concentrations. Pregnant mice were administered increasing doses of ARV combinations by oral gavage, followed by measurement of drug concentrations in the maternal plasma and amniotic fluid. Concentrations of ten different ARVs in maternal plasma and amniotic fluid samples of pregnant mice are presented, with dosing optimization to yield human pregnancy-relevant plasma drug concentrations. We have proposed optimal dosing for different regimen component drugs to achieve human therapeutic plasma levels, so that a clinically relevant standard dosing is established. A review of related ARV pharmacokinetic studies in (pregnant/non-pregnant) rodents and human pregnancy is also shown. We hope these data will inform and encourage the use of mouse pregnancy models in the study of ARV safety/toxicity.

Novel biomarker profiles in experimental aged maternal mice with hypertensive disorders of pregnancy.

Recently, advanced maternal age (AMA) has increased in Western countries because of late marriage and advances in assisted reproductive technology. One major complication of AMA is hypertensive disorders of pregnancy (HDP). While clinical investigations into human AMA have been reported, there has been limited information obtained from basic research. In this investigation, we established the AMA mouse model using aged pregnant ICR mice. We demonstrated that the phenotypes of aged pregnant ICR mice reflect the same characteristics as human AMA. The significant findings of our investigation are as follows: (1) The AMA mouse model manifested the same complication phenotypes of human AMA, including maternal obesity, declining fertility, small for gestational age, and a higher rate of intrauterine fetal death; (2) The AMA mouse model exhibited an increasing systolic blood pressure at late gestation (108.2 ± 7.7 vs. 92.7 ± 5.7 mmHg, P < 0.01) that normalized after delivery similar to human HDP patients; and (3) While HDP and placental dysfunction are complicated, AMA mice and human HDP AMA patients manifested a low serum soluble fms-like tyrosine kinase-1 (sFlt-1) level in late gestation (AMA group vs. control group, mice, 16800.0 ± 10709.5 vs. 26611.9 ± 8702.0 pg/mL, respectively, P < 0.01; human, 8507.6 ± 3298.7 vs. 14816.9 ± 5413.5 pg/mL, respectively, P < 0.05). In conclusion, the aged pregnant mouse model resembled human AMA. The AMA mouse model was complicated with HDP despite the low serum sFlt-1 level. Our findings provide evidence that the serum sFlt-1 level does not necessarily reflect the conventional pathogenesis of HDP in aged human and murine pregnancies and may contribute to the future management of HDP in AMA.

Correction: Cervical HSV-2 infection causes cervical remodeling and increases risk for ascending infection and preterm birth.

[This corrects the article DOI: 10.1371/journal.pone.0188645.].

Animal and Human Tissue Models of Vertical Listeria monocytogenes Transmission and Implications for Other Pregnancy-Associated Infections.

Intrauterine infections lead to serious complications for mother and fetus, including preterm birth, maternal and fetal death, and neurological sequelae in the surviving offspring. Improving maternal and child heath is a global priority. Yet, the development of strategies to prevent and treat pregnancy-related diseases has lagged behind progress made in other medical fields. One of the challenges is finding tractable model systems that replicate the human maternal-fetal interface. Animal models offer the ability to study pathogenesis and host defenses in vivo However, the anatomy of the maternal-fetal interface is highly divergent across species. While many tools are available to study host responses in the pregnant mouse model, other animals have placentas that are more similar to that of humans. Here we describe new developments in animal and human tissue models to investigate the pathogenesis of listeriosis at the maternal-fetal interface. We highlight gaps in existing knowledge and make recommendations on how they can be filled.

The intervention effect of aspirin on a lipopolysaccharide-induced preeclampsia-like mouse model by inhibiting the nuclear factor-κB pathway†

Preeclampsia is a severe pregnancy-related disorder, and patients usually present with high circulating inflammatory factor levels and excessive activation of the nuclear factor-κB (NF-κB) pathway. Administration of aspirin (ASP) is effective for preventing preeclampsia, and thus, we propose that ASP might affect placental function by regulating the NF-κB pathway. Systemic lipopolysaccharide (LPS) (20 μg/kg) was used to induce preeclampsia-like pregnant mouse model, and low-dose ASP (15.2 mg/kg) was administrated. Here, we report significantly increased circulatory expression levels of the proinflammatory cytokines tumor necrosis factor-alpha, interleukin-6, and soluble Fms-related tyrosine kinase-1 in LPS-treated pregnant mice, accompanied by kidney and placental dysfunction. Low-dose ASP treatment significantly reversed the preeclampsia-like phenotype, lowering hypertension, decreasing proteinuria, and ameliorating fetal growth retardation. Moreover, the excessive activation of NF-κB signaling in mice placentae induced by LPS was significantly reduced by ASP. In JEG-3 cells, LPS activated the NF-κB signaling pathway by upregulating the expression of cyclooxygenase-2 (COX-2) and related inflammatory factors, whereas the invasion ability of JEG-3 cells was weakened. However, ASP administration impeded NF-κB signaling activation, downregulated COX-2 and inflammatory factor expression, and rescued trophoblast invasion. This study provides new evidence that low-dose ASP is beneficial for preeclampsia prevention by inhibiting NF-κB and its downstream signaling pathways in trophoblast cells.

Role of chemerin/CMKLR1 in the maintenance of early pregnancy

Chemerin is a cytokine that attracts much attention in the reproductive process. This study aimed to explore the effects of chemerin and its receptor chemokine-like receptor 1 (CMKLR1) on the maintenance of early pregnancy. The expression levels of chemerin and CMKLR1 in the decidua tissues of 20 early normal pregnant women and 20 early spontaneous abortion women were examined by Western blot and real-time polymerase chain reaction analyses. CMKLR1 receptor antagonist (α-NETA) was then intrauterinely injected into normal pregnant mice model to assess its effect on the outcome of pregnancy and the phosphorylation rate of ERK1/2 in decidua tissues.We found that the expression level of chemerin in women who had experienced early spontaneous abortion was lower than in those who had experienced normal early pregnancy (P < 0.01); conversely, CMKLR1 expression was higher in the former than in the latter (P < 0.01). In a pregnant-mouse model, the embryo resorption rate of α-NETA group was higher than that in the negative control group (61.5% vs. 10.8%) (P < 0.001). Compared with the control group, ERK1/2 phosphorylation in decidua tissues decreased in the α-NETA-treated group (P < 0.01). These results suggested that the inhibition of the chemerin/CMKLR1 signaling pathway can lead to the abortion of mouse embryos, and that chemerin/CMKLR1 may play an important role in the maintenance of early pregnancy possibly by regulating ERK1/2 phosphorylation.

Collagen Type 1 Accelerates Healing of Ruptured Fetal Membranes

Preterm premature rupture of membranes (pPROM) is a major cause of preterm birth. Recently, extracellular matrix-directed treatment is applied for wound healing. Here, we used a pregnant mouse model to test the efficacy of collagen type 1 gel for healing of the prematurely ruptured fetal membranes. Although injection of PBS into the ruptured fetal membranes resulted in 40% closure, injection of collagen type 1 improved closure rates to 90% within 72 h. Macrophages of the M2 wound healing phenotype were entrapped in the collagen layer. In primary human amnion mesenchymal cells, collagen type 1 gels activated collagen receptor discoidin domain receptor 2 (DDR2) to induce myosin light chain phosphorylation and migration of injured amnion mesenchymal cells. These findings define the mechanisms for matrix-directed therapeutics for pPROM.

28: Effectiveness of maternal supplementation of a combination of inositols, fucoxanthin and hydroxytyrosol in an obese pregnant mouse model

28: Effectiveness of maternal supplementation of a combination of inositols, fucoxanthin and hydroxytyrosol in an obese pregnant mouse model

Cervical HSV-2 infection causes cervical remodeling and increases risk for ascending infection and preterm birth.

Preterm birth (PTB), or birth before 37 weeks gestation, is the leading cause of neonatal mortality worldwide. Cervical viral infections have been established as risk factors for PTB in women, although the mechanism leading to increased risk is unknown. Using a mouse model of pregnancy, we determined that intra-vaginal HSV2 infection caused increased rates of preterm birth following an intra-vaginal bacterial infection. HSV2 infection resulted in histological changes in the cervix mimicking cervical ripening, including significant collagen remodeling and increased hyaluronic acid synthesis. Viral infection also caused aberrant expression of estrogen and progesterone receptor in the cervical epithelium. Further analysis using human ectocervical cells demonstrated a role for Src kinase in virus-mediated changes in estrogen receptor and hyaluronic acid expression. In conclusion, HSV2 affects proteins involved in tissue hormone responsiveness, causes significant changes reminiscent of premature cervical ripening, and increases risk of preterm birth. Studies such as this improve our chances of identifying clinical interventions in the future.

Biocompatibility assessment of titanium dioxide nanoparticles in mice fetoplacental unit.

As the applications of titanium dioxide nanomaterials (nTiO2 ) are growing with an ever-increasing speed, the hazardous risks of this material have become a major concern. Several recent studies have reported that nTiO2 can cross the placental barrier in pregnant mice and cause neurotoxicity in their offspring. However, the influence of these nanoparticles on the fetoplacental unit during the pregnancy is yet to be studied. The present study reports on the effects of nTiO2 on the anatomical structure of fetal brain and liver in a pregnant mice model. Moreover, changes in the size and weight of the fetus and placenta are investigated as markers of fetal growth. Lastly, the toxicity of nTiO2 in primary brain and liver is quantified. Animals treated with nTiO2 showed a disrupted anatomical structure of the fetal brain and liver. Furthermore, the fetus and placental unit in the mice treated with these nanoparticles were smaller compared to untreated controls. Toxicity analyses revealed that nTiO2 was toxic to the brain and liver cells and the mechanism of cell death was mostly necrosis. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 580-589, 2018.

Decidual Macrophage Functional Polarization during Abnormal Pregnancy due to Toxoplasma gondii: Role for LILRB4.

During gestation, Toxoplasma gondii infection produces a series of complications including stillbirths, abortions, and congenital malformations. The inhibitory receptor, LILRB4, which is mainly expressed by professional antigen-presenting cells (especially macrophages and dendritic cells) may play an important immune-regulatory role at the maternal–fetal interface. To assess the role of LILRB4 during T. gondii infection, LILRB4−/− and T. gondii infected pregnant mouse models were established. Further, human primary-decidual macrophages were treated with anti-LILRB4 neutralizing antibody and then infected with T. gondii. These in vivo and in vitro models were used to explore the role of LILRB4 in T. gondii-mediated abnormal pregnancy outcomes. The results showed that abnormal pregnancy outcomes were more prevalent in LILRB4−/− infected pregnant mice than in wild-type infected pregnant mice. In subsequent experiments, expression levels of LILRB4, M1, and M2 membrane-functional molecules, arginine metabolic enzymes, and related cytokines were assessed in uninfected, infected, LILRB4-neutralized infected, and LILRB4−/− infected models. The results demonstrated T. gondii infection to downregulate LILRB4 on decidual macrophages, which strengthened M1 activation functions and weakened M2 tolerance functions by changing M1 and M2 membrane molecule expression, synthesis of arginine metabolic enzymes, and cytokine secretion profiles. These changes contributed to abnormal pregnancy outcomes. The results of this study provide not only a deeper understanding of the immune mechanisms operational during abnormal pregnancy, induced by T. gondii infection, but also identify potential avenues for therapeutic and preventive treatment of congenital toxoplasmosis.