Technique for the detection of Toxoplasma gondii antigens in mouse urine

Fachado, Alberto; Fonte, Luis; Rojas, Lázara; Alberti, Esteban; Machin, Rodolfo

doi:10.1590/s0074-02761990000100010

Cited by 5 publications

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“…This was more pronounced in compounds 4a and 6. The parasite count results were in accordance with the results obtained from the Co-A test, which served as the best means of diagnosis of recent Toxoplasma infection, and considered a marker of cure (Fachado et al, 1990;Raizman and Neva, 1975). In the present study, no clumping was visible for compounds 4a and 6.…”

Section: Discussionsupporting

confidence: 89%

Synthesis and evaluation of some pyrimidine analogs against toxoplasmosis

et al. 2008

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Toxoplasmosis is a worldwide infection caused by the obligate intracellular protozoan parasite Toxoplasma gondii. Some pyrimidine analogs were synthesized to be evaluated for their antitoxoplasma effects in animal experiments. Results were assessed by studying parasite density, estimation of serum Toxoplasma antigen, studying the ultrastructural changes of the parasite, and the histopathological changes of the affected organs and inhibitory activity for dihydrofolate reductase (DHFR) isolated from Cryptosporidium parvum. The result showed that 4 out of 12 synthesized compounds have promising antitoxoplasma potentials. The animals that received these four compounds showed statistically significant decrease in the mean number of the parasite count in the liver and the spleen when compared to the corresponding control group. Moreover, Toxoplasma antigen was very low or even absent in the serum of animals receiving these compounds. Light microscopic examination of the peritoneal exudates of animals receiving these compounds showed stoppage of movement and deformity in shape of the tachyzoites, whereas scanning electron microscopy revealed that the organisms had lost their crescent shape, with dimples and deep ridges on their surfaces. Very mild histopathological changes were noticed in liver, spleen, and lungs of the groups of animals receiving these compounds in comparison to the other groups. Thus, these compounds proved their effectiveness in eradication of the experimental Toxoplasma infection and inhibition of DHFR from C. parvum.

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Section: Discussionsupporting

confidence: 89%

Synthesis and evaluation of some pyrimidine analogs against toxoplasmosis

et al. 2008

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“…Já foi demonstrado que o coelho mostra-se um bom modelo experimental para o estudo da toxoplasmose (FACHADO et al, 1990;HITT e FILICE, 1992;WATANABE et al,1993;BLACKWELL et al, 1994;PEIFFER et al, 1994;HUNTER et al, 1995), porém * Avaliação variando de: + = pouco freqüente / +++++ = 100% de freqüência ** Grupo I: Inoculado *** Grupo II: Não inoculado (controle) Os sinais clínicos observados, como hipertermia, estertores pulmonares, apatia, taquipnéia e secreção Tabela 6 -Freqüência de alterações microscópicas encontradas em exames histopatológicos das coelhas pertencentes aos grupos inoculados com 1 x 10 4 oocistos de T. gondii (n=9) e não inoculado (n=3). Sharma et al (1973).…”

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Toxoplasmose experimental em coelhas gestantes

Filho¹,

Moraes²,

Moraes³

et al. 2001

Sem. Ci. Agr.

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A toxoplasmose congênita constitui-se numa das formas de maior importância da doença tanto em humanos quanto em animais domésticos, e que se manifesta com maior freqüência durante os dois primeiros trimestres da gestação. Por outro lado, quando adquirida no terceiro trimestre, ocorrem infecções subclínicas no feto ou mesmo ausência de infecção (DESMONTS e COUVREUR, 1974).Em humanos, quando a primoinfecção ocorre durante a gestação, aproximadamente 40% dos fetos se infectam. Dentre estes, uma porcentagem de 15% morrem ou apresentam sintomatologia severa ao nascerem, 19% demonstram sinais clínicos moderados e o restante apresenta infecção subclínica (VITOR, 1994).A transmissão congênita do T. gondii também foi observada em animais utilizados para consumo humano. O T. gondii é descrito como parasito habitual de ovinos, caprinos, suínos e bovinos em várias partes do mundo (VITOR e PINTO, 1991;WARNEKULASURIYA et al., 1998;ESTEBAN et al., 1999). A importância dessa enfermidade nos animais de interesse zootécnico é evidenciada quando são consideradas as perdas econômicas decorrentes, principalmente, de abortos, natimortos e crias debilitadas, devido à transmissão congênita do parasito (LINKLATER e DYSON, 1979;McSPORAN et al., 1985).Apesar da infecção por T. gondii ser relativamente freqüente em coelhos domésticos, em muitos países, a manifestação clínica da toxoplasmose é aparentemente rara nesta espécie (DUBEY et al., 1992).A epidemiologia da toxoplasmose está descrita em lebres selvagens e em coelhos domésticos na Escandinávia e nos EUA (DUBEY et al., 1992), em coelhos selvagens na Alemanha (MANGOLD et al., 1999) e República Tcheca (LITERAK e RYCHLIK, 1999). O coelho constitui-se em reservatório de T. gondii, cuja infecção é mantida graças à transmissão intra-uterina do protozoário (HEJLICEK e LITERAK, 1995). Svoboda e Svobodova (1987) ressaltaram a importância da carne de fígado de coelhos infectados na transmissão da toxoplasmose para cães e gatos.Resumo: Neste ensaio foram comparados aspectos clínicos, obstétricos, imunológicos e patológicos entre coelhas gestantes inoculadas com oocistos de Toxoplasma gondii e coelhas gestantes não inoculadas. Os resultados obtidos demonstram ser a coelha gestante altamente sensível à infecção toxoplásmica. Aborto e óbito foram observados em 100% das coelhas infectadas, antes do término da gestação. Os aumentos máximos de temperatura corporal foram observados entre o quarto e o nono dia pós-inoculação (DPI). Alterações hematológicas também foram observadas. Devido ao caráter hiperagudo decorrente da infecção toxoplásmica, detectou-se resposta imune humoral nas coelhas infectadas somente no 7 o DPI. Os exames anátomo-histopatológicos demonstraram, principalmente, alterações hepáticas, renais e pulmonares. Com base nos dados obtidos no estudo, comprovou-se que coelhas gestantes são extremamente sensíveis a oocistos de T. gondii, apresentando severas alterações em diversos órgãos, assim como aborto e óbito prematuro. Palavras-chave: Toxoplasma gondii, coelhas gestantes, aborto. Abs...

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“…Recently, some researchers have described the use of T. gondii DNA sequence for the diagnosis of toxoplasmosis using the polymerase chain reaction (PCR) (Dupcuy-Camet et al 1993). We have reported the use of coagglutination assay to detect T. gondii antigens in the urine of infected mice (Fachado et al 1990) and also in the urine of AIDS patients (Fachado et al 1992 We have developed the coagglutination technique using urine of AIDS patients with acute cerebral toxoplasmosis. In order to confirm the toxoplasmic origin of the antigens detected by CoAToxo, the immunoblot was employed.…”

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confidence: 99%

Toxoplasma gondii Antigenuria in Patients with Acquired Immune Deficiency Syndrome

Fachado

Fonseca²,

Fonte³

et al. 1997

Mem. Inst. Oswaldo Cruz

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A longitudinal study was performed with sera and urine of patients with acquired immune deficiency syndrome (AIDS), taken before, during and after clinicallyToxoplasma gondii is recognized as an important pathogen during pregnancy and the perinatal period (Remington & Desmont 1983). It has recently gained additional interest as a cause of life threatening opportunistic diseases in immunocompromised individuals, including patients with acquired immune deficiency syndrome (AIDS) and transplant recipients (Postman et al. 1988). Toxoplasmosis is a major opportunistic parasitic disease in AIDS patients. Toxoplasmic encephalitis (TE) can only be diagnosed very late since the classical serological tests based on the detection of anti-T. gondii IgG and IgM are inefficient in these patients (Luft & Remington 1988). More sensitive diagnostic tests for the early diagnosis of TE are needed.The detection of antigens or parasites of T. gondii in different biological fluids is an accurate and rapid diagnosis. Recently, some researchers have described the use of T. gondii DNA sequence for the diagnosis of toxoplasmosis using the polymerase chain reaction (PCR) (Dupcuy-Camet et al. 1993). We have reported the use of coagglutination assay to detect T. gondii antigens in the urine of infected mice (Fachado et al. 1990) and also in the urine of AIDS patients (Fachado et al. 1992 We have developed the coagglutination technique using urine of AIDS patients with acute cerebral toxoplasmosis. In order to confirm the toxoplasmic origin of the antigens detected by CoAToxo, the immunoblot was employed. The results also were correlated with clinical symptoms of toxoplasmosis, response to specific chemotherapy and anatomopathologic exam if the patient died. MATERIALS AND METHODSProduction of the soluble antigenic extract -A soluble antigenic extract was prepared from the RH strain of T. gondii. The trophozoites were taken from peritoneal exudates of mice, three days after infection. The exudate was centrifuged at 650xg for 10 min in 2 ml of 0.1M phosphate buffer saline (PBS), pH 7.2-7.4 to eliminate host cell residues. The supernatant was discarded and the pellet resuspended with 2ml PBS and repeatedly passed through a 26 gauge needle in order to break the macrophages. It was again centrifuged at 160xg during 10 min at 4ºC and the supernatant centrifuged at 650xg during 10 min at 4ºC. The pellet was washed three times in 50 ml PBS. The suspension was then sonicated at 4ºC to obtain the parasites antigens, and centrifuged at 650xg during 2 hr at 4ºC . The protein concentration of the T. gondii antigens suspension was determined by the method of Lowry et al. (1951).Production of antiserum -The anti-Toxoplasma antiserum was prepared by dilution of 0.5 ml of the T. gondii antigens (1.3 mg/ml of protein) in the same volume of PBS. The diluted antigenic solu-

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Technique for the detection of Toxoplasma gondii antigens in mouse urine

Cited by 5 publications

References 0 publications

Synthesis and evaluation of some pyrimidine analogs against toxoplasmosis

Synthesis and evaluation of some pyrimidine analogs against toxoplasmosis

Toxoplasmose experimental em coelhas gestantes

Toxoplasma gondii Antigenuria in Patients with Acquired Immune Deficiency Syndrome

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