Dogs with food allergies and enteropathies may require hydrolyzed diets to prevent or reduce clinical signs, therefore the protein sources used in these diets must be previously characterized and evaluated in healthy dogs. This study aimed to investigate the effects of a hydrolyzed chicken liver powder-based diet (HCLP) versus a poultry by-product meal and bovine meat and bone meal-based diet (Control), on complete blood count (CBC), cytokine, immunoglobulins responses (assessed on days 0, 15, 30 and 45), and fecal microbiota (assessed on day 45) in healthy adult dogs. The CBC did not differ between diets (P>0.05), remaining within reference range. Total plasma IL-4 concentrations were decreased over time independent of the dietary treatment (P<0.001). Total plasma IgA decreased on day 30 compared to days 0 and 45 in dogs fed the control diet (P<0.001). Total plasma IgE concentrations were reduced on days 30 and 45 in dogs fed the control diet, and on days 15 vs 30 and 15 vs 45 in dogs fed HCLP diet (P = 0.001). The 16S rRNA gene sequencing showed similar species richness and abundances of phyla and genera between diets (P>0.05). β-diversity principal coordinate analysis plots demonstrated that HCLP group had a higher similarity than control. Based on our results, healthy adult dogs fed a HCLP based diet maintained normal values for hematological and immunological characteristics, and fecal microbiota after 45 days of feeding.