In broiler production, the litter is reused for consecutives flocks, and it is treated during down time between flocks to reduce its microbial load. Although covering the litter with a plastic canvas is a common litter treatment in the field, there is little scientific information available on its efficacy. The aim of this study was to evaluate the effects of covering broiler litter with a plastic canvas for eight days on litter microbiological, physical, and chemical parameters, and on the intestinal microbiota and immunity of broilers. In the first trial, reused litter from a previous flock was distributed into three treatments, with six replicates each: L1 (negative control, litter free from Salmonella Enteritidis (SE) and Eimeria maxima (EM) and not covered), L2 (positive control, litter with SE and EM, and not covered), and L3 (litter with SE and EM, and covered with plastic canvas for eight days). Litter total bacteria, Enterobacteria, Lactobacillus, SE, and EM counts, and litter pH, temperature, moisture, and ammonia emission were determined on days 1 and 8. In the second trial, broilers were housed on those litters according to the treatments described above, and their intestinal microbiota, gut CD4+ and CD8+ lymphocytes and macrophages, and liver and intestinal proinflammatory interleukin (IFN-γ, IL-1β e IL-18) levels were evaluated on days 14 and 28. A significant reduction of litter bacterial populations was observed in the litter covered with plastic canvas. A significantly higher mRNA IFN-γ gene expression (12.5-fold) was observed in the jejunum and liver of broilers reared on the litter with Enterobacteria counts. No EM reduction was observed in the covered litter. Covering reused broiler litter with plastic canvas reduces initial litter bacterial load as a result of the interaction between physical and chemical parameters.