Pneumonia intersticial crônica em cães naturalmente infectados com Leishmania (Leishmania) chagasi: estudo histopatológico e morfométrico

Eighteen mongrel dogs of unknown age and naturally infected with Leishmania (Leishmania) chagasi, were obtained from the City Hall of Belo Horizonte, Brazil. Four dogs were used as control. Lung samples were obtained and immediately fixed in formalin. The histopathological picture of all lung tissue sections was a chronic and diffuse interstitial pneumonitis. The thickened inter-alveolar septa were characterized by the cellular exudate (mostly macrophages, lymphocytes and plasmocytes) associated with collagen deposition. Morphometric analysis showed greater septal thickness in the infected animals than in controls. In fact, the morphometric study of collagen stained with ammoniac silver confirmed a larger deposition of collagen in the infected animals. The parasitologic method was carried out during the study of the lesions on the slides. However, we did not observe any correlation between the histopathologic and morphometric data and the clinical status of the animals. We conclude that the pulmonary lesions observed in all naturally infected dogs were correlated with the disease and that the morphometric method used was satisfactory for the analysis of septal thickness and of increased collagen deposition, confirming the presence of fibrosis.Neste estudo, foram utilizados dezoito cães naturalmente infectados com Leishmania (Leishmania) chagasi, sem raça e idade definidos, cedidos pelo Centro de Contole de Zoonoses da Cidade de Belo Horizonte. Quatro cães foram utilizados como controle. Durante a necropsia, amostras do pulmão foram coletadas, e imediatamente fixadas em formalina. A histopatologia mostrou principalmente uma pneumonite intersticial crônica, produtiva, difusa e intensa. Os septos inter-alveolares espessados, eram constituídos principalmente por um infiltrado de células inflamatórias, em sua maioria, macrófagos, linfócitos e plasmócitos, além de fibroblastos e fibras colágenas. A análise morfométrica do espessamento septal mostrou diferença significativa entre animais infectados e controle. De fato, a análise morfométrica do colágeno, corado pela prata, confirmou um aumento na deposição de colágeno em todos os animais infectados. O exame parasitológico foi ralizado durante a análise histopatológica, em todos os casos estudados. Contudo neste trabalho, não observamos qualquer correlação entre os achados histopatológicos e morfométricos quando relacionados aos cães em diferentes formas clínicas. Concluímos que as lesões, encontradas nos cães naturalmente infectados, tem correlação com a doença, e que o método de análise morfométrica utilizado apresentou resultados satisfatórios quanto à análise do espessamento septal e ao aumento da deposição de colágeno, confirmando a fibrose

Contribuição ao conhecimento do quadro anatomopatológico do coração na doença de Chagas

Os conhecimentos anatomopatológicos da fase aguda humana da doença de Chagas são baseados, quase que em sua totalidade, em trabalhos realizados há várias décadas. Nos últimos anos, mesmo os patologistas de áreas endêmicas da tripanossomíase cruzi não têm tido oportunidade de necropsiar chagásicos falecidos nessa fase da moléstia. Isto parece importante, visto que é necessário interpretar os achados morfológicos, tendo como base os novos conhecimentos adquiridos pela patologia, bem como, especificamente, colocá-los no contexto das novas idéias que se tem sobre a doença de Chagas. Por estas razões, pareceu-nos de interesse relatar os achados de necropsia por nós realizada, recentemente, em chagásico falecido na fase aguda e chamar a atenção para dados morfológicos cardíacos, que podem contribuir para melhor conhecimento da forma em questão da doença

Comparison of paraffin-embedded skin biopsies from different anatomical regions as sampling methods for detection of Leishmania infection in dogs using histological, immunohistochemical and PCR methods

BACKGROUND: We compared skin biopsy samples from different anatomical regions for detecting Leishmania in dogs, using histological (HE), immunohistochemical (IHC) and polymerase chain reaction (PCR) techniques. RESULTS: The sensitivity was 82.8 percent for PCR, 62.1 percent for IHC and 44.8 percent for HE. These methods do not appear to depend on the clinical status of the animal or the anatomical source of the skin sample; there is no "best region" for any method. However, PCR was more effective than IHC and HE for ear and nose skin samples whereas IHC was better than HE for nose samples. There was weak agreement between PCR and HE for all tissue samples; good agreement between PCR and IHC for ear and abdomen samples, and weak agreement for nose; and optimal agreement between IHC and HE for ear and abdomen and good agreement for nose samples. CONCLUSION: The PCR on ear skin could be the best procedure for diagnosing canine visceral leishmaniasis. The good agreement between PCR and IHC indicates that IHC can be used as an alternative method. Finally, tissue samples from ears, nose and abdomen, particularly ears and nose, are potentially useful for diagnosing canine visceral leishmaniasis independently of the animal's clinical status

Different infective forms trigger distinct immune response in experimental Chagas disease.

Although metacyclic and blood trypomastigotes are completely functional in relation to parasite-host interaction and/or target cell invasion, they differ in the molecules present on the surface. Thus, aspects related to the variability that the forms of T. cruzi interacts with host cells may lead to fundamental implications on the immune response against this parasite and, consequently, the clinical evolution of Chagas disease. We have shown that BT infected mice presented higher levels of parasitemia during all the acute phase of infection. Moreover, the infection with either MT or BT forms resulted in increased levels of total leukocytes, monocytes and lymphocytes, specifically later for MT and earlier for BT. The infection with BT forms presented earlier production of proinflammatory cytokine TNF-α and later of IFN-γ by both T cells subpopulations. This event was accompanied by an early cardiac inflammation with an exacerbation of this process at the end of the acute phase. On the other hand, infection with MT forms result in an early production of IFN-γ, with subsequent control in the production of this cytokine by IL-10, which provided to these animals an immunomodulatory profile in the end of the acute phase. These results are in agreement with what was found for cardiac inflammation where animals infected with MT forms showed intense cardiac inflammation later at infection, with a decrease in the same at the end of this phase. In summary, our findings emphasize the importance of taking into account the inoculums source of T. cruzi, since vectorial or transfusional routes of T. cruzi infection may trigger distinct parasite-host interactions during the acute phase that may influence relevant biological aspects of chronic Chagas disease

Increase of reactive oxygen species by desferrioxamine during experimental Chagas' disease.

Oxidative stress is common in inflammatory processes associated with many diseases including Chagas' disease. The aim of the present study was to evaluate, in a murine model, biomarkers of oxidative stress together with components of the antioxidant system in order to provide an overview of the mechanism of action of the iron chelator desferrioxamine (DFO). The study population comprised 48 male Swiss mice, half of which were treated daily by intraperitoneal injection of DFO over a 35-day period, while half were administered sterile water in a similar manner. On the 14th day of the experiment, 12 DFO-treated mice and an equal number of untreated mice were experimentally infected with Trypanosoma cruzi. Serum concentrations of nitric oxide and superoxide dismutase and hepatic levels of total glutathione, thiobarbituric acid reactive species and protein carbonyl, were determined on days 0, 7, 14 and 21 post-infection. The results obtained revealed that DFO enhances antioxidant activity in the host but also increases oxidative stress, indicating that the mode of action of the drug involves a positive contribution to the host together with an effect that is not beneficial to the parasite

Análise quantitativa das lesões cardíacas na cardiomiopatia chagásica crônica canina

Lesions observed in chronic chagasic cardiopathy frequently produce electrocardiographic alterations and affect cardiac function. Through a computerized morphometrical analysis we quantified the areas occupied by cardiac muscle, connective and adipose tissues in the right atrium of dogs experimentally infected with Trypanosoma cruzi. All of the infected dogs showed chronic myocarditis with variable reduction levels of cardiac muscle, fibrosis and adipose tissue replacement. In the atrial myocardium of dogs infected with Be78 and Be62 cardiac muscle represented 34 and 50%, fibrosis 28 and 32% and adipose tissue 38 and 18%, respectively. The fibrosis observed was both diffuse and focal and mostly intrafascicular, either partially or completely interrupting the path of muscle bundles. Such histological alterations probably contributed to the appearance of electrocardiographic disturbances verified in 10 out 11 dogs which are also common in human chronic chagasic cardiopathy. Fibrosis was the most important microscopic occurrence found since it produces rearrangements of collagen fibers in relation to myocardiocytes which causes changes in anatomical physiognomy and mechanical behavior of the myocardium. These abnormalities can contribute to the appearance of cardiac malfunction, arrythmias and congestive cardiac insufficiency as observed in two of the analyzed dogs. Strain Be78 caused destruction of atrial cardiac muscle higher than that induced by strain Be62.As lesões observadas na cardiopatia chagásica crônica frequentemente produzem alterações eletrocardiográficas e afetam a função cardíaca. Através de uma análise morfométrica computadorizada nós quantificamos as áreas ocupadas por músculo cardíaco, tecido conjuntivo fibroso e tecido adiposo no átrio direito de cães experimentalmente infectados pelo Trypanosoma cruzi. Todos os cães infectados apresentaram miocardite crônica fibrosante com graus variáveis de redução de músculo cardíaco, fibrose e substituição por tecido adiposo. No miocárdio atrial dos cães infectados pelas cepas Be78 e Be62 foram observados 34 e 50% de músculo cardíaco, 28 e 32% de fibrose e, 38 e 18% de tecido adiposo, respectivamente. A fibrose observada era tanto difusa quanto focal e, principalmente intrafascicular interrompendo total ou parcialmente o percurso dos feixes musculares. Tais alterações histológicas provavelmente contribuiram para o surgimento dos distúrbios eletrocardiográficos verificados em 10 dos 11 cães estudados e que são comuns na cardiopatia chagásica crônica humana. De todos os achados microscópicos encontrados, a fibrose foi a mais importante por produzir rearranjos na fibras colágenas em relação aos miocardiócitos, modificando a fisionomia anatômica e o comportamento mecânico do miocárdio. Tais anormalidades estruturais podem contribuir para o surgimento à disfunção cardíaca, arritmias e à insuficiência cardíaca congestiva como verificado em dois cães analisados. A cepa Be78 produziu uma destruição de músculo cardíaco atrial estatisticamente superior à induzida pela cepa Be62

Trypanosoma cruzi: desferrioxamine decreases mortality and parasitemia in infected mice through a trypanostatic effect.

Desferrioxamine (DFO) is a potent iron chelator that is also known to modulate inflammation and act as an efficient antioxidant under normal conditions and under oxidative stress. Many in vitro and in vivo studies have shown the efficacy of DFO in the treatment of viral, bacterial and protozoan infections. DFO is known to reduce the intensity of Trypanosoma cruzi infections in mice even during a course of therapy that is not effective in maintaining anaemia or low iron levels. To further clarify these findings, we investigated the action of DFO on mouse T. cruzi infection outcomes and the direct impact of DFO on parasites. Infected animals treated with DFO (5 mg/animal/day) for 35 days, beginning 14 days prior to infection, presented lower parasitemia and lower cumulative mortality rate. No significant effect was observed on iron metabolism markers, erythrograms, leukograms or lymphocyte subsets. In the rapid method for testing in vivo T. cruzi susceptibility, DFO also induced lower parasitemia. In regard to its direct impact on parasites, DFO slightly inhibited the growth of amastigotes and trypomastigotes in fibroblast culture. Trypan blue staining showed no effects of DFO on parasite viability, and only minor apoptosis in trypomastigotes was observed. Nevertheless, a clear decrease in parasite mobility was detected. In conclusion, the beneficial actions of DFO on mice T. cruzi infection seem to be independent of host iron metabolism and free of significant haematological side effects. Through direct action on the parasite, DFO has more effective trypanostatic than trypanocidal properties

Estudo, ao microscópio óptico e eletrônico, do rim de caes natural e experimentalmente infectados com Leishmania (Leishmania) chagasi

Two naturally infected dogs (male and fema lei from Teófilo Otoni (MG Brazili were maintained for 18 months in our laboratory. Two other dogs, two months old males were infected with 1 x 10(6) promastigotes of MHO BR 70 BH46 Leishinania (Leishmanial chagasi strain, endo venous route, and autopsied after 10 months and two years. The main findings concerning the kidney were: (1) focal or diffuse mesangial glomerulo nephritis with proliferative and enlargement of mesangial cells; (2) increase in thickness of basement membrane with electron dense deposits: (3) chronic interstitial nephritis with intense exudation of plasmocytes: (4) cloud swelling of renal tubules. The authors discuss the probable pathogenetic mechanisms.Os autores estudam os rins de 4 cães infectados com Leishmania (Leishmania) chagasi. Dois animais (um macho e uma fêmea) naturalmente infectados foram sacrificados 18 meses após sua permanência no laboratório. Dois machos foram inoculados por via endovenosa, com lxlO6 promastigotas da cepa MHO/BR/70/BH46 e sacrificados após 18 meses e 2 anos, respectivamente. Em todos os animais os rins estavam lesados. As alterações encontradas foram: (1) glomerulonefrite mesangioproliferativa focal ou difusa, com pronunciada hipertrofia e hiperplasia das células mesangiais e com alargamento da matriz; (2) espessamento da membrana basal com depósitos eletrondensos; (3) nefrite intersticial intertubular crônica com exsudação plasmocitária intensa. (4) degeneração albuminosa dos túbulos renais. Baseados nos achados os autores discutem os prováveis mecanismos patogenéticos