Immunohistochemical detection of Tritrichomonas foetus in experimentally infected mice

National University of La Plata, Eva Perón, Buenos Aires, Argentina
Pesquisa Veterinária Brasileira (Impact Factor: 0.36). 01/2000; 20(1). DOI: 10.1590/S0100-736X2000000100006
Source: DOAJ


The need to intensify knowledge of the pathogenesis of bovine genital trichomoniasis (BGT) led to the use of alternative animal models such as the mouse. Nevertheless, it is necessary to elucidate the dynamics of the infection in this animal species, evaluating different stages of the colonization and evolution of the pathological alterations. The immunohistochemistry (IHC) offers advantages over the routine histopathological staining techniques for the detection of the protozoan in tissues, cellular detritus and inside the macrophages. The goal of the present study was to demonstrate the presence of Tritrichomonas foetus in the reproductive tract of infected mice using an IHC technique. Female BALB/c mice were infected with a suspension of T. foetus by intravaginal route, in the estrum phase, detected by exfoliative vaginal cytology. After 10 weeks, the animals were sacrificed; uterus and vagina were fixed and histologically processed. Some slides were stained with HE. The rest of the slides were processed for IHC. An immunoadsorbed polyclonal serum against T. foetus was used. The avidine-biotine technique (HistoMouse, Zymed[tm]) was employed. The histopathological studies showed a dilation of the uterine glands, presence of macrophages in the lumen of the organ and inner part of the endometrial glands. No T. foetus was identified using this method. The IHQ allowed additionally the identification of the protozoan in the endometrium, endometrial glands, uterine lumen and inside neutrophils and macrophages. The cytological studies stained with IHC showed either isolated T. foetus adhered to epithelial cells or inside macrophages. This technique proves to be a useful tool for the study of the pathogenesis of bovine genital trichomoniasis (BGT) in an experimental model.

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Available from: Enrique L Portiansky, Jul 03, 2014
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    • "Analysis of the kinetics of infection in this murine model required a method of identifying the organism in infected tissue. Traditional histological stains do not allow clear recognition of these protozoa, so immunohistochemistry was employed to detect the membrane and £agellum of T. foetus (Monteavaro et al., 2000). These studies enabled us to identify the parasite within host cells, including macrophages, dendritic cells and the endometrial epithelia. "
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    ABSTRACT: An experimental murine model of bovine genital tritrichomonosis is described. Female mice were inoculated per vaginam with Tritrichomonas foetus and a sample of the study population was killed every 3 days up to 60 days post-infection. Microscopical changes in the reproductive organs were assessed and immunohistochemistry was used to detect T. foetus within these tissues. Lectin histochemistry was used to determine changes in the expression of carbohydrates within the reproductive mucosa. A range of microscopical changes were detected in the uterine endometrium by 10 days post-inoculation and these were associated with the presence of the protozoan. The endometrial changes included endometritis and ulceration, mucosal atrophy and glandular metaplasia, and were similar to those reported in naturally infected cows. Changes in lectin binding were recognized first in the vagina where there was increased binding of Ulex europaeus agglutinin-1 (UEA-1) which was maximal on day 16 post-inoculation. Within the uterus, there was increased binding of soy bean agglutinin (SBA) which was maximal on day 19 post-inoculation, and of peanut agglutinin (PNA) which was maximal on day 16 post-inoculation. These changes in carbohydrate expression parallel the infection kinetics, since they appeared first in the vagina and later in the uterus. The changes may reflect either a host reaction against the infection or the production of enzymes by T. foetus, which act to enhance adhesion and colonization of the genital organs by the organism. The kinetics and pathogenesis of this murine infection are similar to those of the natural bovine disease, suggesting that this model system may be valuable for further studies of this disease.
    Full-text · Article · Feb 2008 · Journal of Comparative Pathology
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    ABSTRACT: The present work reviews aspects related to the diffusion of bovine trichomonosis, emphasizing not only on its morphological and ultrastructural characteristics, but also on the pathogenesis of Tritrichomonas foetus: its effects on bovine genital tract and immune response. Cytotoxical and enzymatic factors and their relationship with antigenic changes provoked by host-parasite interaction are also mentioned. The existing link between the hormonal and immune system during pregnancy and T. foetus' pathogenicity is also discussed in this work. In addition, humoral and mucosal immunity is considered in male and female naturally infected with T. foetus. The immune response against whole cell, membrane and subunit vaccines in vaccinated cattle is regarded/ taken into account too. Recent findings related to diagnosis problems due to non-specific protozan, isolated from preputial smegma of virgin bulls, and the use of other test such as PCR, immunohistochemistry and electronic microscopy, are also included. Finally, recommendations about this disease's control and treatment are also made.
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