Protocolo Experimental in vivo para Testes Antibacterianos e de Potencialização de Antibióticos em Danio rerio (Zebrafish)

• DOI: https://doi.org/10.22533/at.ed.540132510112

• Palavras-chave: Infecção experimental, Resistência microbiana, Ensaios pré-clínicos, Farmacodinâmica experimental.

• Keywords: Experimental infection, Microbial resistance, Pre-clinical trials, Experimental pharmacodynamics.

• Abstract: The increasing microbial resistance has driven the search for compounds of natural or synthetic origin with antimicrobial activity or capable of potentiating the action of antibiotics. In this context, the in vivo model using the Danio rerio fish (zebrafish) has gained prominence due to its high genetic homology with humans and its well-characterized immunological response. Thus, this chapter proposes the standardization of an in vivo experimental protocol for evaluating the antibacterial activity and synergistic potential of natural and synthetic substances, using adult zebrafish as a biological model. The protocol includes the breeding and acclimatization of the animals, preparation of culture media and bacterial inoculum, definition of concentrations of the tested compounds based on the Minimum Inhibitory Concentration (MIC), and intramuscular infection procedures. It also includes oral treatment with antibiotics, evaluated products, or their combinations with antibiotics at subinhibitory concentrations. Furthermore, the protocol presents techniques for animal euthanasia, as well as tissue dissection and plating methods. Therefore, this protocol allows the evaluation of the therapeutic efficacy of compounds in a real biological environment, considering factors of absorption, metabolism, and immune response, which are not possible to observe in in vitro assays.