BEA treatment in infected animals with and without T2D
After two months of infection, treatment with BEA was started in groups of mice with TB or T2D/TB administrated each other day .02 mg/kg by intra-tracheal route to produce a direct effect on the infected lung, and the same amount of BEA administrated by subcutaneous route for systemic distribution promoting hepatic effect. TB group received BEA only by intratracheal instillation. After 30 and 60 days of treatment, groups of six animals were euthanized by exsanguination under anesthesia with 210 mg/kg of intraperitoneal pentobarbital, inside in a cabinet of biosecurity level III. Three left lungs per time were perfused with absolute ethanol, fixed, and prepared for histopathological studies. After eliminating hilar lymph nodes and thymic tissues, five lungs were frozen and kept to −80◦C for bacilli load quantification and three lungs were used to determine the expression of diverse genes by RT-PCR. Two independent experiments were performed. Animals were monitored daily and humanely euthanized if exhibited respiratory insufficiency or substantial weight loss.