Histologic and histomorphometric study of bone repair under acute Trypanosoma cruzi infection in rats (original) (raw)
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RESUMEN El Trypanosoma cruzi (T. cruzi) es un protozoario intracelular que causa Trypanosomoniasis Americana (Enfermedad de Chagas). El objetivo del presente trabajo fue el estudio histo-patológico del efecto de la infección aguda por Trypanosoma cruzi sobre la reparación del tejido óseo. Se utilizaron ratas Wistar macho que fueron asignadas a dos grupos: Grupo Control (GC n =20) y Grupo Experimental (GE n =20). Los animales de ambos grupos, bajo anestesia general intraperitoneal, fueron sometidos a t 0 , a exodoncia del primer molar inferior derecho, en el GE fueron inoculados,a t 0 por vía subcutánea en la región inguinal izquierda con 0.1 mL de 10 5 tripomastigotes de la cepa virulenta Tulahuén de Trypanoso-ma cruzi. A los animales del GC se les administró el volumen equivalente de solución salina por vía subcutánea. A los ani-males de ambos grupos se les practicó la eutanasia a los 15 días. Se resecaron las mandíbulas, se fijaron en solución de formol al 10%, se radiografiaron, ...
Trypanosoma cruzi: experimental parasitism of bone and cartilage
Trypanosoma cruzi causes Chagas' disease, a systemic infection that affects cells of meso-, endo-, and ectodermic origin. However, as far as we know, the presence of T. cruzi stages in bone has not been reported previously, and it has scarcely been investigated in cartilage. We inoculated 7-and 20-day-old (8 and 15 g) NMRI albino mice i.p. with metacyclic trypomastigotes from Rhodnius prolixus used for xenodiagnosis of mice previously infected with mammalian, human, and triatomines isolates, characterized by randomly amplified polymorphic DNA as zymodeme 1 (equivalent to T. cruzi I). Tissular parasitism (quantified according to the number of pseudocysts/50 fields 400×) showed amastigotes, intermediate forms, or trypomastigotes in sternum chondroblasts, osteoblasts, macrophages, and fibroblasts; chondrocyte and osteocyte invasion was rare. All isolates parasitized bone marrow macrophages, with few amastigotes. We observed marked associated myotropism, with or without inflammatory infiltration; there were small numbers of intensely parasitized mononuclear cells in perichondrium and periosteum. We discuss the results in relation to the marked differences of the T. cruzi tropism toward the different types of sternum cells, and, additionally, we outline the possibility of transmitting parasitized bone marrow through transplants. The fact of finding parasite stages in sternum bone and cartilage may be considered important due to the studies on Chagas' disease paleoparasitology that are based on histological and molecular analysis.
In vivo infection by Trypanosoma cruzi: a morphometric study of tissue changes in mice
Parasitology Research, 2013
Nifurtimox and benznidazole, medications currently used for the treatment of the Chagas disease, are not always successful. We determine whether (−)-cubebin and (−)-hinokinin could be used as alternative drugs for the treatment of parasitic infections by Trypanosoma cruzi. To this end, male BALB/c mice were treated with both drugs, and the nuclear parameters (largest diameter, smallest diameter, and perimeter) were determined from slides prepared from the spleen, liver, and heart. The cytotoxicity of the substances was determined after 24-h treatment. Results revealed increased cell nuclei in untreated infected animals as compared to uninfected mice. The values obtained for infected animals treated with (−)-cubebin and (−)-hinokinin were close to those observed for uninfected mice. For the spleen, perimeter values of 10.85 μm (p < 0.01) and 10.90 μm (p<0.05) were obtained for mice treated with (−)-cubebin 50 mg/kg and (−)-hinokinin 20 mg/kg, respectively, whereas untreated infected animals furnished a perimeter of 11.76 μm. As for the liver, perimeter values of 19.06 μm (p<0.01) and 18.61 μm (p<0.001) were achieved for mice treated with (−)-cubebin 50 mg/kg and (−)-hinokinin 20 mg/kg, respectively, whereas a perimeter of 18.54 μm was obtained for untreated infected animals. The cytotoxicity assays demonstrated that (−)-cubebin and (−)-hinokinin does not display toxicity. Therefore, (−)-cubebin and (−)-hinokinin are promising therapeutic agents and could be used in future clinical studies concerning treatment of the Chagas disease. Even if the karyometry is not used frequently, it can complement other methods, such as PCR, and furthermore, it is a simple method which is easily possible to analyze the activity of substances in the tissues of treated infected animals compared to uninfected animals.
Revista da Sociedade Brasileira de Medicina Tropical, 1983
Mice were infected with blood forms of 17 Trypanosoma cruzi strains recently isolated from chronic patients, which were dassified as of low, medium or high virulence on grounds of the prepatent period, parasitemia and mortality at the acute phase. A total of 212 mice were studied after 3, 6, 9 and 12 months of infection. In the chronic phase, intracellular parasites were detected in 11.0%,27.9%and 54.0,% of mice inoculated, respectively, with the low, medium and high virulent strains (r= 0.98, p < 0.005). Heart fibrosis was also related to virulence, affecting 5.7%, 11.6%and30.8% (r = 0.98, p < 0.001) of the mice inoculated with the above strains; a similar relationship was observed between intensity and frequency of the heart inflammatory reaction and the severity of infection at its early stage. Necrotizing arteritis was detected in 12.2% of the inoculated animals and this lesion was related to the infection duration rather than to strain characteristics. Inflammatory lesion...
Molecular and biochemical parasitology, 2018
In the present work, we evaluated the effect of mixed Trypanosoma cruzi infections, studying the biological distribution of the different parasites in blood, heart and skeletal muscle during the acute phase. Albino Swiss mice were infected with different parasite strain/isolates or with a combination of them. The parasites in the different tissues were typified through specific PCR, population variability was analyzed through RFLP studies and parasitological and histopathological parameters were evaluated. We found a predominance of TcII and TcVI in all tissues samples respect to TcV and different parasite populations were found in circulation and in the tissues from the same host. These results verify the distribution of parasites in host tissues from early stages of infection and show biological interactions among different genotypes and populations of T. cruzi.
Research in Veterinary Science, 2014
Trypanosoma brucei brucei Trypanosoma evansi Clastogenenicity Micronucleated polychromatic erythrocyte Bone marrow Micro nucleus assay A B S T R A C T The clastogenic effect of mixed infection of Trypanosoma evansi and Trypanosoma brucei brucei in the bone marrow (BM) cells of Wistar albino rats was investigated. Clastogenic effects were observed in the BM cells using the micronucleus assay. The findings indicate that T. evansi, T. b. brucei and mixed infection with both parasites induced the formation of micronucleated polychromatic erythrocyte (MN-PCEs) in the BM cells significantly (P < 0.05) by 60, 63 and 81 micronuclei/1000 PCE respectively. Mixed infection induced formation of MN-PCEs increase by about 1.33 fold when compared with single infections of T. b. brucei and T. evansi. These data give a preliminary evidence of possible genotoxic effects in trypanosomiasis.
Bulletin of Experimental Biology and Medicine, 2013
Comparative histopathological study and analysis of parasite load in different muscle groups were carried out in BALB/c mice during the acute phase of Chagas disease. Activities of C104 clone of T. cruzi strain TPAP/MX/2002/Albarrada and the parental strain were compared. Panoramic 2D-microscopy imaging of sample surface was used and quantitative analysis of parasitism and pathologic damage was performed. The infection rates in various muscle groups were as follows: myocardium=abdominal muscles=lumbar muscles=femoral muscles←diaphragm for the clone and myocardiumabdominal muscles=lumbar muscles=femoral muscles→ diaphragm for the parental strain.
PLOS Neglected Tropical Diseases, 2015
Oral transmission of Chagas disease has been documented in Latin American countries. Nevertheless, significant studies on the pathophysiology of this form of infection are largely lacking. The few studies investigating oral route infection disregard that inoculation in the oral cavity (Oral infection, OI) or by gavage (Gastrointestinal infection, GI) represent different infection routes, yet both show clear-cut parasitemia and heart parasitism during the acute infection. Herein, BALB/c mice were subjected to acute OI or GI infection using 5x10 4 culture-derived Trypanosoma cruzi trypomastigotes. OI mice displayed higher parasitemia and mortality rates than their GI counterparts. Heart histopathology showed larger areas of infiltration in the GI mice, whereas liver lesions were more severe in the OI animals, accompanied by higher Alanine Transaminase and Aspartate Transaminase serum contents. A differential cytokine pattern was also observed because OI mice presented higher proinflammatory cytokine (IFN-γ, TNF) serum levels than GI animals. Real-time PCR confirmed a higher TNF, IFN-γ, as well as IL-10 expression in the cardiac tissue from the OI group compared with GI. Conversely, TGF-β and IL-17 serum levels were greater in the GI animals. Immunolabeling revealed macrophages as the main tissue source of TNF in infected mice. The high mortality rate observed in the OI mice paralleled the TNF serum rise, with its inhibition by an anti-TNF treatment. Moreover, differences in susceptibility between GI versus OI mice were more clearly related to the host response than to the effect of gastric pH on parasites, since infection in magnesium hydroxide-treated mice showed similar results. Overall, the present study provides conclusive evidence that the initial site of parasite entrance critically affects host immune response and disease outcome. In light of the occurrence of oral Chagas disease outbreaks, our results raise important implications in terms of the current view of the natural disease course and host-parasite relationship.
Comparative histopathological study and analysis of parasite load in different muscle groups were carried out in BALB/c mice during the acute phase of Chagas disease. Activities of C104 clone of T. cruzi strain TPAP/MX/2002/Albarrada and the parental strain were compared. Panoramic 2D-microscopy imaging of sample surface was used and quantitative analysis of parasitism and pathologic damage was performed. The infection rates in various muscle groups were as follows: myocardium=abdominal muscles=lumbar muscles=femoral muscles←diaphragm for the clone and myocardiumabdominal muscles=lumbar muscles=femoral muscles→ diaphragm for the parental strain.
BioMed Research International, 2014
A murine model was used to study the histopathological aspects and cytokine expression levels in skeletal muscle provoked by the infection with Mexican TcI strains. BALB/c mice were inoculated with the virulent Querétaro strain and the nonvirulent Ninoa strain. Parasite numbers were counted in blood and skeletal muscle at different times post-infection, and real time-PCR expression levels of the cytokines IL-12, IL-4, IL-10, IFN-, and TNF-were evaluated. In the acute phase of infection, a high parasitic load, both in blood and skeletal muscle, was detected. The histopathological analyses showed an exacerbated inflammation and granulomatous-like infiltrate with the Querétaro strain. Interestingly, extensive calcification areas were observed in the skeletal muscle surrounded by inflammatory infiltrates. TNF-and IL-10 expression exhibited a significant increase at the peak of infection. In summary, Querétaro strain, a Mexican TcI strain, is virulent enough to induce high inflammation and calcification in skeletal muscle of the hind limbs, which could be related to high expression levels of TNF-.