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      Serological and molecular detection of Neospora caninum and Toxoplasma gondii in human umbilical cord blood and placental tissue samples

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          Abstract

          Neosporosis primarily affects cattle and dogs and is not currently considered a zoonotic disease. Toxoplasmosis is a zoonosis with a worldwide distribution that is asymptomatic in most cases, but when acquired during pregnancy, it can have serious consequences. The seropositivity rates determined by the indirect fluorescent antibody test for Neospora caninum ( N. caninum) and Toxoplasma gondii ( T. gondii) were 24.3% (49 samples) and 26.8% (54 samples), respectively. PCR positivity for N. caninum was observed in two samples of cord blood (1%) using the Nc5 and ITS1 gene, positivity for T. gondii was observed in 16 samples using the primer for the B1 gene (5.5% positivity in cord blood and 2.5% positivity in placental tissue). None of the samples showed structures characteristic of tissue cysts or inflammatory infiltrate on histopathology. Significant associations were observed only between N. caninum seropositivity and the presence of domestic animals (p = 0.039) and presence of dogs (p = 0.038) and between T. gondii seropositivity and basic sanitation (p = 0.04). This study obtained important findings regarding the seroprevalence and molecular detection of N. caninum and T. gondii in pregnant women; however, more studies are necessary to establish a correlation between risk factors and infection.

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          Most cited references57

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          Toxoplasma invasion of mammalian cells is powered by the actin cytoskeleton of the parasite.

          Toxoplasma gondii is an obligate intracellular parasite that invades a wide range of vertebrate host cells. We demonstrate that invasion is critically dependent on actin filaments in the parasite, but not the host cell. Invasion into cytochalasin D (CD)-resistant host cells was blocked by CD, while parasite mutants invaded wild-type host cells in the presence of drug. CD resistance in Toxoplasma was mediated by a point mutation in the single-copy actin gene ACT1. Transfection of the mutant act1 allele into wild-type Toxoplasma conferred motility and invasion in the presence of CD. We conclude that host cell invasion by Toxoplasma, and likely by related Apicomplexans, is actively powered by an actin-based contractile system in the parasite.
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            Direct and sensitive detection of a pathogenic protozoan, Toxoplasma gondii, by polymerase chain reaction.

            We applied the polymerase chain reaction to detection of the pathogenic protozoan Toxoplasma gondii based on our identification of a 35-fold-repetitive gene (the B1 gene) as a target. Using this procedure, we were able to amplify and detect the DNA of a single organism directly from a crude cell lysate. This level of sensitivity also allowed us to detect the B1 gene from purified DNA samples containing as few as 10 parasites in the presence of 100,000 human leukocytes. This is representative of the maximal cellular infiltration (10(5)/ml) in 1 ml of cerebrospinal fluid obtained from patients with toxoplasmic encephalitis. The B1 gene is present and conserved in all six T. gondii strains tested to date, including two isolates from patients with acquired immunodeficiency syndrome. No signal was detected by using this assay and DNAs from a variety of other organisms, including several which might be found in the central nervous system of an immunocompromised host. This combination of sensitivity and specificity should make detection of the B1 gene based on polymerase chain reaction amplification a very useful method for diagnosis of toxoplasmosis both in immunocompromised hosts and in congenitally infected fetuses.
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              Advances in the life cycle of Toxoplasma gondii.

              J.P. Dubey (1998)
              This paper reviews recent studies on the life cycle of Toxoplasma gondii. Tachyzoites, bradyzoites, and sporozoites are the three infectious stages of T. gondii. Humans and animals become infected mainly by ingesting bradyzoites or oocytes. After ingestion, both bradyzoites and sporozoites convert to tachyzoites inside tissues. The conversion of tachyzoites to bradyzoites and bradyzoites to tachyzoites is of biological and clinical significance because bradyzoites are less susceptible to chemotherapy and reactivation of bradyzoites to tachyzoites is considered the cause of fatal toxoplasmosis in AIDS patients. Of all the methods currently available to assess stage conversion of T. gondii, feeding infective stages to cats is the most reliable method. Felidae, the definitive hosts of T. gondii excrete oocysts 3-10 days after ingesting tissue cysts/bradyzoites, > or = 18 days after ingesting oocysts, and > or = 13 days after ingesting tachyzoites.
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                Author and article information

                Contributors
                renato.andreotti@embrapa.br
                Journal
                Sci Rep
                Sci Rep
                Scientific Reports
                Nature Publishing Group UK (London )
                2045-2322
                3 June 2020
                3 June 2020
                2020
                : 10
                : 9043
                Affiliations
                [1 ]ISNI 0000 0001 2163 5978, GRID grid.412352.3, Programa de Pós-Graduação em Doenças Infecciosas e Parasitárias- Universidade Federal de Mato Grosso do Sul (UFMS), ; Campo Grande, MS Brasil
                [2 ]ISNI 0000 0004 0541 873X, GRID grid.460200.0, Laboratório de Biologia Molecular do Carrapato, Departamento de Sanidade Animal, Embrapa Gado de Corte, ; Campo Grande, MS Brasil
                [3 ]ISNI 0000 0004 0462 7943, GRID grid.441738.d, Professor do Curso de Medicina Veterinária do Centro Universitário da Grande Dourados-UNIGRAN, ; Dourados, Brasil
                [4 ]ISNI 0000 0004 0541 873X, GRID grid.460200.0, Bolsista de Pós-Doutorado- FUNDAPAM, Laboratório de Biologia Molecular do Carrapato, Departamento de Sanidade Animal, Embrapa Gado de Corte, ; Campo Grande, MS Brasil
                [5 ]Laboratório de Toxonologia e Plantas Medicinais-Uniderp Agrárias, Campo Grande, Brasil
                [6 ]ISNI 0000 0004 0541 873X, GRID grid.460200.0, Empresa Brasileira de Pesquisa Agropecuária, Embrapa Gado de Corte, ; Campo Grande, MS Brasil
                Article
                65991
                10.1038/s41598-020-65991-1
                7271125
                32493968
                4c2f3c60-311d-4633-a19e-b37ec7abc951
                © The Author(s) 2020

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 3 December 2019
                : 13 May 2020
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                © The Author(s) 2020

                Uncategorized
                molecular biology,diseases
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                molecular biology, diseases

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