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The aim of this study was to evaluate the diffuse intralobular fibrosis in dogs naturally infected with Leishmania (Leishmania) chagasi. One hundred five infected animals with positive serologic tests for Leishmania were divided into two clinical groups: 69 symptomatic animals and 36 asymptomatic. Special staining with Gomori, Heidenhain, Silver, and Picrosirius Red was applied to characterize fibrilopoesis. The tissue parasite load was measured by immunohistochemistry and associated histomorphometric analyses. Intralobular fibrosis was observed in all dogs, and more collagen deposition was confirmed in the infected animals than in the controls by these histomorphometric studies. There were significant differences among the distinct clinical groups. In fact, symptomatic dogs showed an increased collagen deposition in the liver compared with asymptomatic ones. A peculiar diffuse intralobular fibrosis, where the collagen fibers encircled small groups of hepatocyte(s), was observed in two cases (1.9%).

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The present data show a fast and efficient biological sample processing method for the extraction of thiamine (vitamin B1) and its mono-(TMP) and di-(TDP) phosphate esters from hippocampus, thalamus and prefrontal cortex (PFC) and blood sample of the rodents. In addition, using the hippocampus and standards of these three compounds we validated an isocratic fluorescence HPLC procedure for a simultaneous detection of them in a single chromatogram within a total run time of about 12 min. Reproducibility for TDP, TMP and B1 was 2.66%, 4.50% and 7.43% (intraday) and 37.54%, 25.39% and 25.87% (interday), respectively. Recovery assays were between 96.0% and 101.7%. The calibration curves were linear and the concentrations of the three compounds, all in nanomolar range, were determined in the brain areas and in the blood samples. When compared to the current methods in the literature, this new method provides information on essential variables, such as linearity range and limit of detection, reproducibility and stability of thiamine, TMP and TDP in rat brain samples. The present data on sample processing and B1 and its phosphate ester level determinations are the first to be validated using hippocampus samples of rats.

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