Brucellosis presents a notable challenge due to the intracellular persistence and adaptability of Brucella species within macrophages, complicating effective treatment strategies. Nanotechnology-based approaches predominantly CSNPs offer promising strategy for targeted drug delivery and improved therapeutic efficiency. CS and its derivatives serve as advantageous vaccine adjuvants and delivery vehicles, attributed to their significant positive charge and capacity to traverse biological barriers while binding to both drugs and cells. CSNPs have applications in mucosal delivery, especially in nasal administration, serving as mucosal vaccine adjuvants in various species including cattle, rabbits, mice, pigs, and chickens. CSNPs loaded with Brucella abortus (B. abortus) malate dehydrogenase (Mdh) induced pro-inflammatory cytokine production and systemic Immunoglobulin A (IgA) in mice through intranasal administration, demonstrating efficacy in eliciting strong immune responses, particularly against B. abortus infections. The evaluation of antimicrobial properties of CSNPs containing gentamicin through minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) assays demonstrated the efficacy of this formulation in eradicating both B. abortus and Brucella melitensis (B. melitensis) strains. The intraperitoneal administration of N-trimethyl chitosan/urease NPs enhances specific immune responses and increases protective efficacy against B. abortus and B. melitensis. The findings highlight the potential of CSNPs as a novel platform for brucellosis treatment, offering advantages compared to conventional treatment methods.