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1.
J Orthop Surg Res ; 19(1): 386, 2024 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-38951811

RESUMO

BACKGROUND: Bone defects, resulting from substantial bone loss that exceeds the natural self-healing capacity, pose significant challenges to current therapeutic approaches due to various limitations. In the quest for alternative therapeutic strategies, bone tissue engineering has emerged as a promising avenue. Notably, excretory proteins from Toxoplasma gondii (TgEP), recognized for their immunogenicity and broad spectrum of biological activities secreted or excreted during the parasite's lifecycle, have been identified as potential facilitators of osteogenic differentiation in human bone marrow mesenchymal stem cells (hBMSCs). Building on our previous findings that TgEP can enhance osteogenic differentiation, this study investigated the molecular mechanisms underlying this effect and assessed its therapeutic potential in vivo. METHODS: We determined the optimum concentration of TgEP through cell cytotoxicity and cell proliferation assays. Subsequently, hBMSCs were treated with the appropriate concentration of TgEP. We assessed osteogenic protein markers, including alkaline phosphatase (ALP), Runx2, and Osx, as well as components of the BMP/Smad signaling pathway using quantitative real-time PCR (qRT-PCR), siRNA interference of hBMSCs, Western blot analysis, and other methods. Furthermore, we created a bone defect model in Sprague-Dawley (SD) male rats and filled the defect areas with the GelMa hydrogel, with or without TgEP. Microcomputed tomography (micro-CT) was employed to analyze the bone parameters of defect sites. H&E, Masson and immunohistochemical staining were used to assess the repair conditions of the defect area. RESULTS: Our results indicate that TgEP promotes the expression of key osteogenic markers, including ALP, Runx2, and Osx, as well as the activation of Smad1, BMP2, and phosphorylated Smad1/5-crucial elements of the BMP/Smad signaling pathway. Furthermore, in vivo experiments using a bone defect model in rats demonstrated that TgEP markedly promoted bone defect repair. CONCLUSION: Our results provide compelling evidence that TgEP facilitates hBMSC osteogenic differentiation through the BMP/Smad signaling pathway, highlighting its potential as a therapeutic approach for bone tissue engineering for bone defect healing.


Assuntos
Diferenciação Celular , Células-Tronco Mesenquimais , Osteogênese , Ratos Sprague-Dawley , Transdução de Sinais , Toxoplasma , Células-Tronco Mesenquimais/metabolismo , Osteogênese/fisiologia , Humanos , Animais , Transdução de Sinais/fisiologia , Diferenciação Celular/fisiologia , Masculino , Toxoplasma/fisiologia , Ratos , Proteínas Smad/metabolismo , Proteínas de Protozoários/metabolismo , Proteínas Morfogenéticas Ósseas/metabolismo , Células Cultivadas
2.
BMJ Open ; 14(6): e084582, 2024 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-38950992

RESUMO

BACKGROUND: Toxoplasmosis is a zoonotic parasitic disease caused by Toxoplasma gondii (T. gondii). It has a wide host range and is capable of vertical transmission in pregnant women, which may lead to undesirable pregnancy outcomes such as congenital malformations, miscarriage, premature birth and stillbirth. This study investigated the seroprevalence of T. gondii infection among pregnant women attending the antenatal clinic at Namwala District Hospital in Southern Zambia. METHODS: This was a cross-sectional study where blood was collected, and the serum was tested for Toxoplasma IgG and IgM. A questionnaire was administered to participants on demographic characteristics and risk factors. Data were entered in Microsoft Excel and exported to STATA version 14 for analysis. RESULTS: A total of 401 women were enrolled in the study from 3 March to 5 August 2021. The seroprevalence of Toxoplasma IgG was 4.2% (n=17), while the seroprevalence of Toxoplasma IgM was 0.7% (n=3). The median age was 27 (IQR: 24-30) years, and a larger proportion had primary-level education (n=223, 55.6%). The majority (81.6%) of the women were married. None of the risk factors investigated in this study were significant for T. gondii infection. CONCLUSION: There was a low seroprevalence of T. gondii infection among pregnant women in the Namwala district of Southern Province, Zambia, and regular screening may not be warranted in this population. Continued research on toxoplasmosis is recommended to understand its epidemiology across Zambia.


Assuntos
Anticorpos Antiprotozoários , Imunoglobulina M , Complicações Parasitárias na Gravidez , Toxoplasma , Toxoplasmose , Humanos , Feminino , Zâmbia/epidemiologia , Estudos Transversais , Estudos Soroepidemiológicos , Adulto , Gravidez , Toxoplasmose/epidemiologia , Toxoplasmose/sangue , Fatores de Risco , Toxoplasma/imunologia , Adulto Jovem , Imunoglobulina M/sangue , Anticorpos Antiprotozoários/sangue , Complicações Parasitárias na Gravidez/epidemiologia , Complicações Parasitárias na Gravidez/sangue , Imunoglobulina G/sangue , Cuidado Pré-Natal
3.
Zhongguo Xue Xi Chong Bing Fang Zhi Za Zhi ; 36(3): 304-309, 2024 Jun 07.
Artigo em Chinês | MEDLINE | ID: mdl-38952318

RESUMO

OBJECTIVE: To investigate the development and dynamic changes of cysts in the brain of mice following infection with different forms of Toxoplasma gondii, so as to provide insights into for toxoplasmosis prevention and control. METHODS: ICR mice at ages of 6 to 8 weeks, each weighing 20 to 25 g, were intraperitoneally injected with tachyzoites of the T. gondii PRU strain at a dose of 1 × 105 tachyzoites per mouse, orally administered with cysts at a dose of 20 oocysts per mouse or oocysts at a dose of 200 oocysts per mouse for modeling chronic T. gondii infection in mice, and the clinical symptoms and survival of mice were observed post-infection. Mice were orally infected with T. gondii cysts at doses of 10 (low-dose group), 20 (medium-dose group), 40 cysts per mouse (high-dose group), and the effect of different doses of T. gondii infections on the number of cysts was examined in the mouse brain. Mice were orally administered with T. gondii cysts at a dose of 20 cysts per mouse, and grouped according to gender (female and male) and time points of infections (20, 30, 60, 90, 120, 150, 180 days post-infection), and the effects of gender and time points of infections on the number of cysts was examined in the mouse brain. In addition, mice were divided into the tachyzoite group (Group T), the first-generation cyst group (Group C1), the second-generation cyst group (Group C2), the third-generation cyst (Group C3) and the fourth-generation cyst group (Group C4). Mice in the Group T were intraperitoneally injected with T. gondii tachyzoites at a dose of 1 × 105 tachyzoites per mouse, and the cysts were collected from the mouse brain tissues 30 days post-infection, while mice in the Group C1 were orally infected with the collected cysts at a dose of 30 cysts per mouse. Continuous passage was performed by oral administration with cysts produced by the previous generation in mice, and the effect of continuous passage on the number of cysts was examined in the mouse brain. RESULTS: Following infection with T. gondii tachyzoites, cysts and oocysts in mice, obvious clinical symptoms were observed on days 6 to 13 and mice frequently died on days 7 to 12. The survival rates of mice were 67.0%, 87.0% and 53.0%, and the mean numbers of cysts were (516.0 ± 257.2), (1 203.0 ± 502.0) and (581.0 ± 183.1) in the mouse brain (F = 11.94, P < 0.01) on day 30 post-infection with T. gondii tachyzoites, cysts and oocysts, respectively, and the numbers of cysts in the brain tissues were significantly lower in mice infected with T. gondii tachyzoites and oocysts than in those infected with cysts (all P values < 0.01). The survival rates of mice were 87.0%, 87.0% and 60.0%, and the mean numbers of cysts were (953.0 ± 355.5), (1 084.0 ± 474.3) and (1 113.0 ± 546.0) in the mouse brain in the low-, medium- and high-dose groups on day 30 post-infection, respectively (F = 0.42, P > 0.05). The survival rates of male and female mice were 73.0% and 80.0%, and the mean numbers of cysts were (946.4 ± 411.4) and (932.1 ± 322.4) in the brain tissues of male and female mice, respectively (F = 1.63, P > 0.05). Following continuous passage, the mean numbers of cysts were (516.0 ± 257.2), (1 203.0 ± 502.0), (896.8 ± 332.3), (782.5 ± 423.9) and (829.2 ± 306.0) in the brain tissues of mice in the T, C1, C2, C3 and C4 groups, respectively (F = 4.82, P < 0.01), and the number of cysts was higher in the mouse brain in Group 1 than in Group T (P < 0.01). Following oral administration of 20 T. gondii cysts in mice, cysts were found in the moues brain for the first time on day 20 post-infection, and the number of cysts gradually increased over time, peaked on days 30 and 90 post-infection and then gradually decreased; however, the cysts were still found in the mouse brain on day 180 post-infection. CONCLUSIONS: There is a higher possibility of developing chronic T. gondii infection in mice following infection with cysts than with oocysts or tachyzoites and the most severe chronic infection is seen following infection with cysts. The number of cysts does not correlate with the severity of chronic T. gondii infection, and the number of cysts peaks in the mouse brain on days 30 and 90 post-infection.


Assuntos
Encéfalo , Camundongos Endogâmicos ICR , Toxoplasma , Toxoplasmose Animal , Animais , Camundongos , Feminino , Masculino , Encéfalo/parasitologia , Doença Crônica , Toxoplasmose Animal/parasitologia , Toxoplasma/fisiologia , Toxoplasmose/parasitologia , Modelos Animais de Doenças
4.
Zhongguo Xue Xi Chong Bing Fang Zhi Za Zhi ; 36(3): 279-285, 2024 Jun 13.
Artigo em Chinês | MEDLINE | ID: mdl-38952314

RESUMO

OBJECTIVE: To prepare and characterize the mouse polyclonal antibody against the dense granule protein 24 (GRA24) of Toxoplasma gondii, and explore its preliminary applications. METHODS: The GRA24 coding sequences of different T. gondii strains were aligned using the MEGA-X software, and the dominant peptide of the GRA24 protein was analyzed with the Protean software. The base sequence encoding this peptide was amplified using PCR assay and ligated into the pET-28a vector, and the generated GRA24 truncated protein was transformed into Escherichia coli BL21. After induction by isopropyl-beta-D-thiogalactopyranoside (IPTG), the expression and purification of the recombinant GRA24 protein was analyzed using sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE). BALB/c mice were immunized by subcutaneous injection with the purified recombinant GRA24 truncated protein to generate the polyclonal antibody, and the titer of the polyclonal antibody was measured using enzyme linked immunosorbent assay (ELISA). The specificity of the polyclonal antibody was tested using Western blotting, and the intracellular localization of the polyclonal antibody was investigated using immunofluorescence assay (IFA). RESULTS: SDS-PAGE showed successful construction of the recombinant expression plasmid, and Coomassie brilliant blue staining showed the generation of the high-purity recombinant GRA24 truncated protein. ELISA measured that the titer of the polyclonal antibody against the GRA24 truncated protein was higher than 1:208 400, and Western blotting showed that the polyclonal antibody was effective to recognize the endogenous GRA24 proteins of different T. gondii strains and specifically recognize the recombinant GRA24 truncated protein. Indirect IFA showed that the GRA24 protein secreted 16 hour following T. gondii invasion in host cells. CONCLUSIONS: The polyclonal antibody against the T. gondii GRA24 protein has been successfully prepared, which has a widespread applicability, high titers and a high specificity. This polyclonal antibody is available for Western blotting and IFA, which provides the basis for investigating the function of the GRA24 protein.


Assuntos
Anticorpos Antiprotozoários , Camundongos Endogâmicos BALB C , Proteínas de Protozoários , Toxoplasma , Animais , Toxoplasma/imunologia , Toxoplasma/genética , Proteínas de Protozoários/imunologia , Proteínas de Protozoários/genética , Camundongos , Anticorpos Antiprotozoários/imunologia , Feminino , Proteínas Recombinantes/imunologia , Especificidade de Anticorpos , Antígenos de Protozoários/imunologia , Antígenos de Protozoários/genética
5.
PLoS One ; 19(7): e0292408, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38950025

RESUMO

Co-infections are a common reality but understanding how the immune system responds in this context is complex and can be unpredictable. Heligmosomoides bakeri (parasitic roundworm, previously Heligmosomoides polygyrus) and Toxoplasma gondii (protozoan parasite) are well studied organisms that stimulate a characteristic Th2 and Th1 response, respectively. Several studies have demonstrated reduced inflammatory cytokine responses in animals co-infected with such organisms. However, while general cytokine signatures have been examined, the impact of the different cytokine producing lymphocytes on parasite control/clearance is not fully understood. We investigated five different lymphocyte populations (NK, NKT, γδ T, CD4+ T and CD8+ T cells), five organs (small intestine, Peyer's patches, mesenteric lymph nodes, spleen and liver), and 4 cytokines (IFN©, IL-4, IL-10 and IL-13) at two different time points (days 5 and 10 post T. gondii infection). We found that co-infected animals had significantly higher mortality than either single infection. This was accompanied by transient and local changes in parasite loads and cytokine profiles. Despite the early changes in lymphocyte and cytokine profiles, severe intestinal pathology in co-infected mice likely contributed to early mortality due to significant damage by both parasites in the small intestine. Our work demonstrates the importance of taking a broad view during infection research, studying multiple cell types, organs/tissues and time points to link and/or uncouple immunological from pathological findings. Our results provide insights into how co-infection with parasites stimulating different arms of the immune system can lead to drastic changes in infection dynamics.


Assuntos
Coinfecção , Citocinas , Nematospiroides dubius , Toxoplasma , Animais , Coinfecção/imunologia , Coinfecção/parasitologia , Toxoplasma/imunologia , Camundongos , Citocinas/metabolismo , Nematospiroides dubius/imunologia , Infecções por Strongylida/imunologia , Infecções por Strongylida/parasitologia , Infecções por Strongylida/mortalidade , Toxoplasmose/imunologia , Toxoplasmose/mortalidade , Toxoplasmose/complicações , Feminino , Toxoplasmose Animal/imunologia , Toxoplasmose Animal/mortalidade , Toxoplasmose Animal/parasitologia , Baço/imunologia , Baço/patologia , Baço/parasitologia , Carga Parasitária , Tecido Linfoide/imunologia , Tecido Linfoide/patologia , Tecido Linfoide/parasitologia
6.
PLoS One ; 19(7): e0306245, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38950027

RESUMO

INTRODUCTION: Toxoplasma gondii can cause symptomatic toxoplasmosis in immunodeficient hosts, including in people living with human immunodeficiency virus (PLWH), mainly because of the reactivation of latent infection. We assessed the prevalence of toxoplasmosis and its associated risk factors in PLWH in the Asia-Pacific region using data from the TREAT Asia Human Immunodeficiency Virus (HIV) Observational Database (TAHOD) of the International Epidemiology Databases to Evaluate AIDS (IeDEA) Asia-Pacific. METHODS: This study included both retrospective and prospective cases of toxoplasmosis reported between 1997 and 2020. A matched case-control method was employed, where PLWH diagnosed with toxoplasmosis (cases) were each matched to two PLWH without a toxoplasmosis diagnosis (controls) from the same site. Sites without toxoplasmosis were excluded. Risk factors for toxoplasmosis were analyzed using conditional logistic regression. RESULTS: A total of 269/9576 (2.8%) PLWH were diagnosed with toxoplasmosis in 19 TAHOD sites. Of these, 227 (84%) were reported retrospectively and 42 (16%) were prospective diagnoses after cohort enrollment. At the time of toxoplasmosis diagnosis, the median age was 33 years (interquartile range 28-38), and 80% participants were male, 75% were not on antiretroviral therapy (ART). Excluding 63 out of 269 people without CD4 values, 192 (93.2%) had CD4 ≤200 cells/µL and 162 (78.6%) had CD4 ≤100 cells/µL. By employing 538 matched controls, we found that factors associated with toxoplasmosis included abstaining from ART (odds ratio [OR] 3.62, 95% CI 1.81-7.24), in comparison to receiving nucleoside reverse transcriptase inhibitors plus non-nucleoside reverse transcriptase inhibitors, HIV exposure through injection drug use (OR 2.27, 95% CI 1.15-4.47) as opposed to engaging in heterosexual intercourse and testing positive for hepatitis B virus surface antigen (OR 3.19, 95% CI 1.41-7.21). Toxoplasmosis was less likely with increasing CD4 counts (51-100 cells/µL: OR 0.41, 95% CI 0.18-0.96; 101-200 cells/µL: OR 0.14, 95% CI 0.06-0.34; >200 cells/µL: OR 0.02, 95% CI 0.01-0.06), when compared to CD4 ≤50 cells/µL. Moreover, the use of prophylactic cotrimoxazole was not associated with toxoplasmosis. CONCLUSIONS: Symptomatic toxoplasmosis is rare but still occurs in PLWH in the Asia-Pacific region, especially in the context of delayed diagnosis, causing advanced HIV disease. Immune reconstitution through early diagnosis and ART administration remains a priority in Asian PLWH.


Assuntos
Infecções por HIV , Toxoplasmose , Humanos , Masculino , Fatores de Risco , Adulto , Feminino , Toxoplasmose/epidemiologia , Toxoplasmose/complicações , Infecções por HIV/epidemiologia , Infecções por HIV/complicações , Ásia/epidemiologia , Estudos Retrospectivos , Estudos de Casos e Controles , Pessoa de Meia-Idade , Prevalência , Estudos Prospectivos , Toxoplasma
7.
Parasit Vectors ; 17(1): 284, 2024 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-38956725

RESUMO

BACKGROUND: Toxoplasma gondii infection affects a significant portion of the global population, leading to severe toxoplasmosis and, in immunocompromised patients, even death. During T. gondii infection, disruption of gut microbiota further exacerbates the damage to intestinal and brain barriers. Therefore, identifying imbalanced probiotics during infection and restoring their equilibrium can regulate the balance of gut microbiota metabolites, thereby alleviating tissue damage. METHODS: Vimentin gene knockout (vim-/-) mice were employed as an immunocompromised model to evaluate the influence of host immune responses on gut microbiota balance during T. gondii infection. Behavioral experiments were performed to assess changes in cognitive levels and depressive tendencies between chronically infected vim-/- and wild-type (WT) mice. Fecal samples were subjected to 16S ribosomal RNA (rRNA) sequencing, and serum metabolites were analyzed to identify potential gut probiotics and their metabolites for the treatment of T. gondii infection. RESULTS: Compared to the immunocompetent WT sv129 mice, the immunocompromised mice exhibited lower levels of neuronal apoptosis and fewer neurobehavioral abnormalities during chronic infection. 16S rRNA sequencing revealed a significant decrease in the abundance of probiotics, including several species of Lactobacillus, in WT mice. Restoring this balance through the administration of Lactobacillus murinus and Lactobacillus gasseri significantly suppressed the T. gondii burden in the intestine, liver, and brain. Moreover, transplantation of these two Lactobacillus spp. significantly improved intestinal barrier damage and alleviated inflammation and neuronal apoptosis in the central nervous system. Metabolite detection studies revealed that the levels of various Lactobacillus-related metabolites, including indole-3-lactic acid (ILA) in serum, decreased significantly after T. gondii infection. We confirmed that L. gasseri secreted much more ILA than L. murinus. Notably, ILA can activate the aromatic hydrocarbon receptor signaling pathway in intestinal epithelial cells, promoting the activation of CD8+ T cells and the secretion of interferon-gamma. CONCLUSION: Our study revealed that host immune responses against T. gondii infection severely disrupted the balance of gut microbiota, resulting in intestinal and brain damage. Lactobacillus spp. play a crucial role in immune regulation, and the metabolite ILA is a promising therapeutic compound for efficient and safe treatment of T. gondii infection.


Assuntos
Lesões Encefálicas , Microbioma Gastrointestinal , Camundongos Knockout , Toxoplasma , Animais , Camundongos , Toxoplasma/imunologia , Lesões Encefálicas/imunologia , Probióticos/administração & dosagem , Encéfalo/imunologia , Lactobacillus , Modelos Animais de Doenças , Hospedeiro Imunocomprometido , Toxoplasmose/imunologia , RNA Ribossômico 16S/genética , Masculino , Intestinos/imunologia
8.
Parasit Vectors ; 17(1): 299, 2024 Jul 10.
Artigo em Inglês | MEDLINE | ID: mdl-38987795

RESUMO

BACKGROUND: Toxoplasma gondii infection causes adverse pregnancy outcomes by affecting the expression of immunotolerant molecules in decidual immune cells. Galectin-9 (Gal-9) is widely expressed in decidual macrophages (dMφ) and is crucial for maintaining normal pregnancy by interacting with the immunomodulatory protein T-cell immunoglobulin and mucin domain-containing molecule 3 (Tim-3). However, the effects of T. gondii infection on Gal-9 expression in dMφ, and the impact of altered Gal-9 expression levels on the maternal-fetal tolerance function of decidual natural killer (dNK) cells, are still unknown. METHODS: Pregnancy outcomes of T. gondii-infected C57BL/6 and Lgals9-/- pregnant mice models were recorded. Expression of Gal-9, c-Jun N-terminal kinase (JNK), phosphorylated JNK (p-JNK), and Forkhead box protein O1 (FOXO1) was detected by western blotting, flow cytometry or immunofluorescence. The binding of FOXO1 to the promoter of Lgals9 was determined by chromatin immunoprecipitation-polymerase chain reaction (ChIP-PCR). The expression of extracellular signal-regulated kinase (ERK), phosphorylated ERK (p-ERK), cAMP-response element binding protein (CREB), phosphorylated CREB (p-CREB), T-box expressed in T cells (T-bet), interleukin 10 (IL-10), and interferon gamma (IFN-γ) in dNK cells was assayed by western blotting. RESULTS: Toxoplasma gondii infection increased the expression of p-JNK and FOXO1 in dMφ, resulting in a reduction in Gal-9 due to the elevated binding of FOXO1 with Lgals9 promoter. Downregulation of Gal-9 enhanced the phosphorylation of ERK, inhibited the expression of p-CREB and IL-10, and promoted the expression of T-bet and IFN-γ in dNK cells. In the mice model, knockout of Lgals9 aggravated adverse pregnancy outcomes caused by T. gondii infection during pregnancy. CONCLUSIONS: Toxoplasma gondii infection suppressed Gal-9 expression in dMφ by activating the JNK/FOXO1 signaling pathway, and reduction of Gal-9 contributed to dysfunction of dNK via Gal-9/Tim-3 interaction. This study provides new insights for the molecular mechanisms of the adverse pregnancy outcomes caused by T. gondii.


Assuntos
Galectinas , Células Matadoras Naturais , Macrófagos , Camundongos Endogâmicos C57BL , Toxoplasma , Toxoplasmose , Animais , Feminino , Gravidez , Galectinas/genética , Galectinas/metabolismo , Camundongos , Células Matadoras Naturais/imunologia , Macrófagos/imunologia , Toxoplasma/imunologia , Toxoplasmose/imunologia , Decídua/imunologia , Camundongos Knockout , Receptor Celular 2 do Vírus da Hepatite A/genética , Receptor Celular 2 do Vírus da Hepatite A/metabolismo , Resultado da Gravidez , Proteína Forkhead Box O1/genética , Proteína Forkhead Box O1/metabolismo
9.
Cell Mol Life Sci ; 81(1): 294, 2024 Jul 09.
Artigo em Inglês | MEDLINE | ID: mdl-38977495

RESUMO

The obligate intracellular parasite Toxoplasma gondii causes life-threatening toxoplasmosis to immunocompromised individuals. The pathogenesis of Toxoplasma relies on its swift dissemination to the central nervous system through a 'Trojan Horse' mechanism using infected leukocytes as carriers. Previous work found TgWIP, a protein secreted from Toxoplasma, played a role in altering the actin cytoskeleton and promoting cell migration in infected dendritic cells (DCs). However, the mechanism behind these changes was unknown. Here, we report that TgWIP harbors two SH2-binding motifs that interact with tyrosine phosphatases Shp1 and Shp2, leading to phosphatase activation. DCs infected with Toxoplasma exhibited hypermigration, accompanying enhanced F-actin stress fibers and increased membrane protrusions such as filopodia and pseudopodia. By contrast, these phenotypes were abrogated in DCs infected with Toxoplasma expressing a mutant TgWIP lacking the SH2-binding motifs. We further demonstrated that the Rho-associated kinase (Rock) is involved in the induction of these phenotypes, in a TgWIP-Shp1/2 dependent manner. Collectively, the data uncover a molecular mechanism by which TgWIP modulates the migration dynamics of infected DCs in vitro.


Assuntos
Movimento Celular , Células Dendríticas , Proteína Tirosina Fosfatase não Receptora Tipo 11 , Proteína Tirosina Fosfatase não Receptora Tipo 6 , Proteínas de Protozoários , Toxoplasma , Toxoplasma/metabolismo , Proteína Tirosina Fosfatase não Receptora Tipo 11/metabolismo , Proteína Tirosina Fosfatase não Receptora Tipo 11/genética , Células Dendríticas/metabolismo , Células Dendríticas/parasitologia , Animais , Proteína Tirosina Fosfatase não Receptora Tipo 6/metabolismo , Proteínas de Protozoários/metabolismo , Proteínas de Protozoários/genética , Humanos , Camundongos , Quinases Associadas a rho/metabolismo , Toxoplasmose/metabolismo , Toxoplasmose/parasitologia , Toxoplasmose/patologia , Camundongos Endogâmicos C57BL
10.
Int J Mol Sci ; 25(13)2024 Jun 25.
Artigo em Inglês | MEDLINE | ID: mdl-39000057

RESUMO

Toxoplasma gondii, an important opportunistic pathogen, underscores the necessity of developing novel therapeutic drugs and identifying new drug targets. Our findings indicate that the half-maximal inhibitory concentrations (IC50) of KU60019 and CP466722 (abbreviated as KU and CP) against T. gondii are 0.522 µM and 0.702 µM, respectively, with selection indices (SI) of 68 and 10. Treatment with KU and CP affects the in vitro growth of T. gondii, inducing aberrant division in the daughter parasites. Transmission electron microscopy reveals that KU and CP prompt the anomalous division of T. gondii, accompanied by cellular enlargement, nuclear shrinkage, and an increased dense granule density, suggesting potential damage to parasite vesicle transport. Subsequent investigations unveil their ability to modulate the expression of certain secreted proteins and FAS II (type II fatty acid synthesis) in T. gondii, as well as including the dot-like aggregation of the autophagy-related protein ATG8 (autophagy-related protein 8), thereby expediting programmed death. Leveraging DARTS (drug affinity responsive target stability) in conjunction with 4D-Label-free quantitative proteomics technology, we identified seven target proteins binding to KU, implicated in pivotal biological processes such as the fatty acid metabolism, mitochondrial ATP transmission, microtubule formation, and Golgi proteins transport in T. gondii. Molecular docking predicts their good binding affinity. Furthermore, KU has a slight protective effect on mice infected with T. gondii. Elucidating the function of those target proteins and their mechanism of action with ATM kinase inhibitors may potentially enhance the treatment paradigm for toxoplasmosis.


Assuntos
Proteínas Mutadas de Ataxia Telangiectasia , Inibidores de Proteínas Quinases , Toxoplasma , Toxoplasma/efeitos dos fármacos , Toxoplasma/enzimologia , Animais , Camundongos , Proteínas Mutadas de Ataxia Telangiectasia/metabolismo , Proteínas Mutadas de Ataxia Telangiectasia/antagonistas & inibidores , Inibidores de Proteínas Quinases/farmacologia , Inibidores de Proteínas Quinases/química , Toxoplasmose/tratamento farmacológico , Toxoplasmose/parasitologia , Humanos , Proteínas de Protozoários/metabolismo , Proteínas de Protozoários/antagonistas & inibidores , Feminino
11.
Rom J Ophthalmol ; 68(2): 198-201, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39006325

RESUMO

Objective: To present a case of ocular toxoplasmosis. Materials and methods: A sixteen-year-old female patient presented to our clinic with complaints regarding decreased vision in her right eye (BCVA 0.5), starting five days before the exam. Her anamnestic data revealed a previous history of ocular toxoplasmosis in her left eye. OCT scans of the inner retina identified a huge cystic space, located posterior to the inner line, off the outer plexiform layer, with a small amount of hyperreflective foci. Other features of OCT included membranous-like structures on inner borders and elongation and splitting of the inner segment/outer segment junction. In later stages, beginning signs of retinitis and scaring could be observed. Results: The patient was treated with sulfamethoxazole/trimethoprim and prednisolone. After two weeks, total regression occurred and visual acuity and OCT remained stable for 6 months (BCVA 1.0). Discussion: Ocular toxoplasmosis can cause significant vision loss due to retinitis and scarring. Following treatment with sulfamethoxazole/trimethoprim and prednisolone, the patient's condition improved significantly and her visual acuity remained stable. Conclusion: On clinical examination and using OCT, rare morphological cystoid spaces (CS) can be identified as huge outer retina cysts (HORC), which are pathognomonic for posterior uveitis. Abbreviations: HORC = huge outer retinal cyst, OCT = optical coherence tomography, BCVA = best corrected visual acuity, CS = cyst space, OPL = outer plexiform layer, HRF = hyper reflective foci, RPE = retinal pigment epithelium, IS = inner segment, OS = outer segment, ERM = epiretinal membrane, PORT = punctate outer retinal toxoplasmosis, ELM = external limiting membrane.


Assuntos
Tomografia de Coerência Óptica , Toxoplasmose Ocular , Acuidade Visual , Humanos , Feminino , Toxoplasmose Ocular/diagnóstico , Toxoplasmose Ocular/tratamento farmacológico , Toxoplasmose Ocular/parasitologia , Tomografia de Coerência Óptica/métodos , Adolescente , Combinação Trimetoprima e Sulfametoxazol/uso terapêutico , Infecções Oculares Parasitárias/diagnóstico , Infecções Oculares Parasitárias/parasitologia , Infecções Oculares Parasitárias/tratamento farmacológico , Angiofluoresceinografia/métodos , Prednisolona/uso terapêutico , Retina/parasitologia , Retina/patologia , Glucocorticoides/uso terapêutico , Fundo de Olho , Toxoplasma/isolamento & purificação
12.
Parasite ; 31: 38, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38995111

RESUMO

In total, 901 dairy cow sera and data were collected from 51 farms in Nakhon Pathom, Ratchaburi and Kanchanaburi provinces (Western Region of Thailand). Serum samples were processed via the multispecies ELISA method to detect IgG antibodies against Toxoplasma gondii infection. The results demonstrated that the calculated true prevalence was 1.48% (95% CI, 0.64-2.75%) for the individual-level and 29.41% (95% CI, 18.71-43%) for the farm-level. The univariate risk factor analysis showed that the number of total owned cats, the presence of stray cats, and the frequency of cleaning per day were significant factors (p < 0.2). These three factors were subjected to logistic regression analysis, and the results revealed that the frequency of cleaning farms per day was a potential risk factor for T. gondii-seropositive farms (OR = 2.745, 95% CI, 1.15-8.69, p = 0.02). The frequency of cleaning might increase the T. gondii oocyst distribution within the barn area, thus increasing the possibility of infection. Our findings show that T. gondii continues to circulate in the dairy cow population in the western part of Thailand. The presence of cats on farms was not found to be associated with T. gondii infection, but the high frequency of cleaning the floor was, and contributed to the potential risk of infection.


Title: Prévalence et facteurs de risque de l'infection à Toxoplasma gondii chez les bovins laitiers de la région occidentale de la Thaïlande. Abstract: Au total, 901 sérums de vaches laitières et des données ont été collectés dans 51 fermes des provinces de Nakhon Pathom, Ratchaburi et Kanchanaburi (région occidentale de la Thaïlande). Les échantillons de sérum ont été traités via la méthode ELISA multi-espèces pour détecter les anticorps IgG contre l'infection à Toxoplasma gondii. Les résultats ont démontré que la prévalence réelle calculée était de 1,48 % (IC à 95 %, 0,64­2,75 %) au niveau individuel et de 29,41 % (IC à 95 %, 18,71­43 %) au niveau des exploitations. L'analyse factorielle a montré que le nombre total de chats possédés, la présence de chats errants et la fréquence quotidienne de nettoyage étaient des facteurs significatifs (p < 0,2). Ces trois facteurs ont été soumis à une analyse de régression logistique et les résultats ont révélé que la fréquence quotidienne de nettoyage des exploitations était un facteur de risque potentiel pour les exploitations séropositives à T. gondii (OR = 2,745, IC à 95 % = 1,15­8,69, p = 0,02). La fréquence du nettoyage pourrait favoriser la répartition des oocystes de T. gondii dans les étables, augmentant ainsi le risque d'infection. Nos résultats indiquent que T. gondii continue de circuler dans la population de vaches laitières de l'ouest de la Thaïlande. La présence de chats dans les fermes n'a pas été associée à l'infection à T. gondii, mais la fréquence élevée du nettoyage du sol l'était et contribuait au risque potentiel d'infection.


Assuntos
Anticorpos Antiprotozoários , Doenças dos Bovinos , Ensaio de Imunoadsorção Enzimática , Imunoglobulina G , Toxoplasma , Toxoplasmose Animal , Animais , Bovinos , Tailândia/epidemiologia , Toxoplasmose Animal/epidemiologia , Fatores de Risco , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/parasitologia , Toxoplasma/imunologia , Anticorpos Antiprotozoários/sangue , Feminino , Gatos , Estudos Soroepidemiológicos , Imunoglobulina G/sangue , Ensaio de Imunoadsorção Enzimática/veterinária , Indústria de Laticínios , Prevalência , Doenças do Gato/epidemiologia , Doenças do Gato/parasitologia , Modelos Logísticos
13.
PLoS Negl Trop Dis ; 18(7): e0012286, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38959260

RESUMO

BACKGROUND: Habitat modification and land use changes impact ecological interactions and alter the relationships between humans and nature. Mexico has experienced significant landscape modifications at the local and regional scales, with negative effects on forest cover and biological biodiversity, especially in the Yucatan peninsula in southeastern Mexico. Given the close relationship between landscape modification and the transmission of zoonotic and vector-borne diseases, it is essential to develop criteria for identifying priority zoonoses in the south of the country. METHODOLOGY/PRINCIPAL FINDINGS: We reviewed 165 published studies on zoonotic and vector-borne diseases in the region (2015-2024). We identified the most frequent vectors, reservoirs, and hosts, the most prevalent infections, and the factors associated with transmission risk and the anthropogenic landscape modification in urban, rural, ecotone, and sylvatic habitats. The most relevant pathogens of zoonotic risk included Trypanosoma cruzi, arboviruses, Leishmania, Rickettsia, Leptospira, and Toxoplasma gondii. Trypanosoma cruzi was the vector-borne agent with the largest number of infected vertebrate species across habitats, while Leishmania and arboviruses were the ones that affected the greatest number of people. Dogs, cats, backyard animals, and their hematophagous ectoparasites are the most likely species maintaining the transmission cycles in human settlements, while rodents, opossums, bats, and other synanthropic animals facilitate connection and transmission cycles between forested habitats with human-modified landscapes. Pathogens displayed different prevalences between the landscapes, T. cruzi, arbovirus, and Leptospira infections were the most prevalent in urban and rural settlements, whereas Leishmania and Rickettsia had similar prevalence across habitats, likely due to the diversity and abundance of the infected vectors involved. The prevalence of T. gondii and Leptospira spp. may reflect poor hygiene conditions. Additionally, results suggest that prevalence of zoonotic and vector-borne diseases is higher in deforested areas and agricultural aggregates, and in sites with precarious health and infrastructure services. CONCLUSIONS: Some hosts, vectors, and transmission trends of zoonotic and vector-borne diseases in the YP are well known but others remain poorly recognized. It is imperative to reinforce practices aimed at increasing the knowledge, monitoring, prevention, and control of these diseases at the regional level. We also emphasize the need to perform studies on a larger spatio-temporal scale under the socio-ecosystem perspective, to better elucidate the interactions between pathogens, hosts, vectors, environment, and sociocultural and economic aspects in this and many other tropical regions.


Assuntos
Doenças Transmitidas por Vetores , Zoonoses , Animais , Humanos , Zoonoses/transmissão , Zoonoses/epidemiologia , Doenças Transmitidas por Vetores/transmissão , Doenças Transmitidas por Vetores/epidemiologia , Prevalência , México/epidemiologia , Ecossistema , Trypanosoma cruzi/isolamento & purificação , Vetores de Doenças , Reservatórios de Doenças/microbiologia , Leptospira/isolamento & purificação , Leptospira/genética , Leptospira/classificação , Doença de Chagas/transmissão , Doença de Chagas/epidemiologia , Toxoplasma , Arbovírus/fisiologia , Leishmania/isolamento & purificação , Leishmaniose/transmissão , Leishmaniose/epidemiologia
14.
Turkiye Parazitol Derg ; 48(2): 128-132, 2024 Jun 30.
Artigo em Inglês | MEDLINE | ID: mdl-38958491

RESUMO

Toxoplasma gondii (T. gondii) is an obligate intracellular, zoonotic protozoan parasite of interest to physicians and veterinarians with its highly complex structure. It is known to infect about one-third of the world's population. Since it is a zoonotic disease, it is necessary to keep the animal population under control in order to prevent human exposure. Many studies have been conducted on the detection of T. gondii and it has been determined that there are three clonal groups consisting of types 1, 2, 3. Developments in molecular studies have led to changes in the taxonomy and new developments in parasitic diseases. It has helped in diagnosis, treatment, development of antiparasitic drugs and research on resistance. They also provided research on vaccine studies, genetic typing and phylogenetics of parasitic diseases. Conventional polymerase chain reaction (PCR), real-time PCR and genotyping studies conducted today increase our knowledge about T. gondii. Methods such as B1, SAG1, SAG2, GRA1, 529-bp repeat element, OWP genes and 18S rRNAs are mostly used in PCR, and methods such as MS, MLST, PCR-RFLP, RAPD-PCR and HRM are used in genotyping. Toxoplasmosis is a disease that is within the framework of the concept of one health and must attract attention, has not yet been eradicated in the world and needs joint studies for humans, animals and ecosystems to be eradicated. This can only be possible by establishing interdisciplinary groups, conducting surveys and training.


Assuntos
Toxoplasma , Toxoplasmose Animal , Toxoplasmose , Toxoplasma/genética , Toxoplasma/classificação , Animais , Humanos , Toxoplasmose/parasitologia , Toxoplasmose Animal/parasitologia , Toxoplasmose Animal/diagnóstico , Zoonoses/parasitologia , Genótipo
15.
Biomed Environ Sci ; 37(6): 647-660, 2024 Jun 20.
Artigo em Inglês | MEDLINE | ID: mdl-38988115

RESUMO

Toxoplasma gondii( T. gondii or Tg), is an obligatory intracellular parasite with humans as its intermediate hosts. In recent years, significant correlations between T. gondii infection and schizophrenia have been reported, including the possible mediating mechanisms. Currently, mechanisms and hypotheses focus on central neurotransmitters, immunity, neuroinflammation, and epigenetics; however, the exact underlying mechanisms remain unclear. In this article, we review the studies related to T. gondii infection and schizophrenia, particularly the latest research progress. Research on dopamine (DA) and other neurotransmitters, the blood-brain barrier, inflammatory factors, disease heterogeneity, and other confounders is also discussed. In addition, we also summarized the results of some new epidemiological investigations.


Assuntos
Esquizofrenia , Toxoplasma , Toxoplasmose , Esquizofrenia/parasitologia , Esquizofrenia/etiologia , Humanos , Toxoplasmose/complicações , Toxoplasmose/epidemiologia , Toxoplasmose/parasitologia , Animais
16.
J Infect Dev Ctries ; 18(6): 964-971, 2024 Jun 30.
Artigo em Inglês | MEDLINE | ID: mdl-38991003

RESUMO

INTRODUCTION: Since the Syrian Civil War began in 2011, the official number of refugees under temporary protection in Turkiye is reported to be 3,522,036 in 2023. Most of the Syrians living outside the refugee camps have worse conditions in terms of access to healthcare centers and social opportunities, compared to those living in camps. The Sanliurfa province hosts the third highest number of Syrians (370,291) in Turkiye. There are no data about the seroprevalence of Toxoplasma gondii (T. gondii), rubella (rub), or cytomegalovirus (CMV) among Syrian refugees in Sanliurfa. We aimed to investigate the seroprevalence of T. gondii, rub, and CMV infections among female Syrian refugees of reproductive age (15-49 years) living in Sanliurfa province. METHODOLOGY: A cross-sectional study was conducted in different districts of Sanliurfa. A total of 460 households were selected using the probability sampling method. One married female Syrian refugee aged between 15 and 49 years, was chosen in each household, leading to a sample size of 410 female Syrian refugees. The seropositivity of T. gondii, CMV, and rub IgM and IgG in blood samples were analyzed using enzyme immunoassays (Abbott Architect, Illinois, USA). RESULTS: The seropositivity rates of T. gondii, CMV, and rubella IgM and IgG were 4.4% and 59.8%; 3.9%; and 99%; and 1.9%, and 99.5%, respectively. CONCLUSIONS: A screening program should be implemented for T. gondii, CMV, and rub infections for Syrian refugees. Seronegative women should be vaccinated against rub and educated about the transmission and preventive routes of toxoplasmosis and CMV infection.


Assuntos
Infecções por Citomegalovirus , Refugiados , Rubéola (Sarampo Alemão) , Toxoplasmose , Humanos , Feminino , Refugiados/estatística & dados numéricos , Adulto , Estudos Soroepidemiológicos , Toxoplasmose/epidemiologia , Adolescente , Adulto Jovem , Rubéola (Sarampo Alemão)/epidemiologia , Síria/epidemiologia , Síria/etnologia , Pessoa de Meia-Idade , Estudos Transversais , Infecções por Citomegalovirus/epidemiologia , Turquia/epidemiologia , Toxoplasma/imunologia , Anticorpos Antivirais/sangue , Citomegalovirus/imunologia , Citomegalovirus/isolamento & purificação , Imunoglobulina M/sangue
17.
Vet Parasitol Reg Stud Reports ; 53: 101052, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-39025537

RESUMO

The feline population is extensive in urban areas worldwide, comprising stray and domestic cats. Cats, acting as reservoirs, can transmit various zoonotic organisms to humans, which can cause significant public health issues. We evaluated the seroprevalence of zoonotic pathogens in stray cats in an urban area of northeast Spain (the city of Zaragoza) to assess potential risks to human health. A total of 88 sampled cats (52 females and 36 males) underwent antibody evaluation using the indirect immunofluorescence technique. Seroprevalence rates were determined for IgG antibodies to Bartonella henselae (36.3%), Toxoplasma gondii (31.8%), Rickettsia felis (14.7%), Rickettsia typhi (9%), and Leishmania infantum (10.2%). Our results confirmed the presence in stray cats of antibodies against all those pathogens, indicating that they all circulate in the feline population in Zaragoza. Male cats exhibited a higher predisposition to T. gondii, whereas females showed an increased likelihood of contracting B. henselae. This difference may be attributed to distinct behaviors according to sex. Our findings underscore the importance of maintaining and intensifying surveillance coupled with preventive measures against zoonotic pathogens in cats. They highlight the need for comprehensive control strategies designed to mitigate public health risks associated with feline populations.


Assuntos
Bartonella henselae , Doenças do Gato , Toxoplasma , Toxoplasmose Animal , Zoonoses , Animais , Gatos , Espanha/epidemiologia , Estudos Soroepidemiológicos , Doenças do Gato/epidemiologia , Doenças do Gato/parasitologia , Doenças do Gato/microbiologia , Masculino , Feminino , Toxoplasma/imunologia , Toxoplasma/isolamento & purificação , Bartonella henselae/imunologia , Bartonella henselae/isolamento & purificação , Toxoplasmose Animal/epidemiologia , Zoonoses/epidemiologia , Zoonoses/parasitologia , Anticorpos Antiprotozoários/sangue , Leishmania infantum/imunologia , Leishmania infantum/isolamento & purificação , Rickettsia typhi/isolamento & purificação , Rickettsia typhi/imunologia , Anticorpos Antibacterianos/sangue , Rickettsia felis/isolamento & purificação , Humanos
18.
Vet Parasitol Reg Stud Reports ; 53: 101063, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-39025547

RESUMO

Seroprevalence studies on cats are essential for monitoring the occurrence of Toxoplasma gondii infection. The present research investigated anti-T. gondii antibodies, risk factors, clinical signs, hematology and serum biochemistry in cats from different regions of Rio de Janeiro. An overall 18.7% (17/91) of the cats were seroreactive, and age was associated with increased chances of seroprevalence of anti-T. gondii antibodies. Clinical signs, hematology and serum biochemistry parameters did not help achieve an antemortem diagnosis of cat toxoplasmosis. The parasite circulates in cats from three major regions of Rio de Janeiro, and the present data set will contribute to future epidemiological studies in this endemic state of Brazil.


Assuntos
Anticorpos Antiprotozoários , Doenças do Gato , Toxoplasma , Toxoplasmose Animal , Gatos , Animais , Brasil/epidemiologia , Estudos Soroepidemiológicos , Doenças do Gato/epidemiologia , Doenças do Gato/parasitologia , Toxoplasmose Animal/epidemiologia , Toxoplasmose Animal/parasitologia , Toxoplasma/imunologia , Fatores de Risco , Anticorpos Antiprotozoários/sangue , Feminino , Masculino
19.
Vet Parasitol Reg Stud Reports ; 53: 101072, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-39025550

RESUMO

Toxoplasma gondii is one of the world's most widespread polyxenic protozoan parasites that affect all warm-blooded animals, including humans. This survey aims to study, for the first time in Algeria, the seroprevalence of Toxoplasma infection in zoo animals. The study included eight animal species of which 54 serum samples were collected from 30 Australian goats (Capra hircus), four bulls (Bos taurus), one dromedary (Camelus dromedarius), three cuffed sheep (Ammotragus lervia), seven donkeys (Equus asinus), one pony (Equus ferus), four bearded horses (Equus ferus caballus) and four rabbits (Oryctolagus cuniculus). The presence of antibodies to T. gondii was determined using the ID Screen® Toxoplasmosis Indirect Multispecies ELISA kit (IDVet, Grabels, France). A total of 8/54 (14.8%) samples were seropositive, including 5/28 (17.9%) males and 3/26 (11.5%) females. The seroprevalence was 6.7%, 50%, 25% and 75% in Capra hircus, Bos Taurus, Equus ferus caballus, and Oryctolagus cuniculus, respectively. No cases were observed in Camelus dromedarius, Ammotragus lervia, Equus asinus, and Equus ferus. This study indicates, for the first time in Algeria, the seroprevalence of T. gondii in zoo animals.


Assuntos
Animais de Zoológico , Anticorpos Antiprotozoários , Toxoplasma , Toxoplasmose Animal , Animais , Toxoplasmose Animal/epidemiologia , Toxoplasmose Animal/parasitologia , Argélia/epidemiologia , Estudos Soroepidemiológicos , Animais de Zoológico/parasitologia , Toxoplasma/imunologia , Toxoplasma/isolamento & purificação , Feminino , Masculino , Anticorpos Antiprotozoários/sangue , Cabras , Bovinos , Ensaio de Imunoadsorção Enzimática/veterinária , Cavalos/parasitologia , Coelhos/parasitologia , Ovinos
20.
Rev Bras Parasitol Vet ; 33(2): e006024, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38985053

RESUMO

Neospora caninum is a major cause of reproductive loss in cattle worldwide as it leads to abortion and animal repositioning. Although Toxoplasma gondii does not cause a reproductive problem in cattle, consuming raw or uncooked beef poses the risk of transmission. This study aimed to evaluate the occurrence of anti-N. caninum and anti-T. gondii antibodies in dairy cattle in the West and Northwest regions of São Paulo State, Brazil. A total of 653 serum samples from dairy cows were analyzed using an indirect immunofluorescence assay (IFA). Epidemiological data from the farms were associated with the serological results of the animals by logistic regression based on the presence of antibodies. The frequencies of the antibodies against N. caninum and T. gondii were 41.6% (272/653) and 11.5% (75/653), respectively. A statistically significant association was observed between: the serum anti-N. caninum antibodies and breed, history of food supplementation for calves, introduction of outside animals that later presented reproductive problems, and history of reproductive problems by the trimester of gestation. The present study highlights the importance of neosporosis in dairy cattle in the study regions and that the inclusion of this parasite in the investigation of animals with reproductive disorders is important.


Assuntos
Anticorpos Antiprotozoários , Doenças dos Bovinos , Coccidiose , Neospora , Toxoplasma , Toxoplasmose Animal , Animais , Bovinos , Neospora/imunologia , Brasil/epidemiologia , Coccidiose/veterinária , Coccidiose/epidemiologia , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/parasitologia , Toxoplasmose Animal/epidemiologia , Toxoplasmose Animal/diagnóstico , Fatores de Risco , Estudos Soroepidemiológicos , Toxoplasma/imunologia , Feminino , Anticorpos Antiprotozoários/sangue , Indústria de Laticínios , Técnica Indireta de Fluorescência para Anticorpo/veterinária
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