Adaptation and development of an experimental model for studies of inflammation in Danio rerio (zebrafish)

• Grant number: 13/25971-9
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: February 01, 2014
• End date: November 30, 2015
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Pathology
• Principal Investigator: Marco Antonio de Andrade Belo
• Grantee: Melque Ferrari de Oliveira
• Host Institution: Universidade Camilo Castelo Branco (UNICASTELO). Campus Descalvado. Descalvado , SP, Brazil

Abstract

Summary of Proposed WorkThis study aims to adapt and develop an experimental protocol for studying the acute inflammatory reaction in Danio rerio ( " zebrafish " ) , induced by inoculation of Aeromonas hydrophila aerocistite . 60 zebrafish ( Danio rerio ) , from the same spawning , which will be packaged in 6 aquaria ( n = 10 ) , with a capacity of 100L of water each , supplied with running water devoid of chlorine will be used . After acclimatization period of 15 days, the fish will be anesthetized in aqueous solution of benzocaine ( 1:10,000 ) for delivery of 10¼L of inoculum of A. hydrophila living in the swim bladder in concentration 3 x 109 CFU . Six , 12 , 18 , 24 , 36 and 48 hours after inoculation , euthanasia ten fish per period is accomplished by deep anesthesia in aqueous solution of benzocaine ( 1:500 ) . The swim bladder is washed thoroughly with PBS containing 0.09% EDTA for collection of exudate and an aliquot of this volume will be transferred to a Neubauer chamber to count the total inflammatory cells by light microscopy. For differential counting of thrombocytes , lymphocytes , macrophages, and granulocytes is placed a small aliquot of the exudate for histological slide over an extended embodiment , subsequent to dye fixation with May - Grunwald -Giemsa - Wright . Although our laboratory have experience in the art as with other teleost pacu ( Piaractus mesopotamicus ) and tilapia ( Oreochromis niloticus ), the administration of micro - inoculum in the swim bladder to the small size of the zebrafish , the standardization of the anesthesia and the determination of kinetic cell response in the inflamed focus on the six proposed periods represent the challenge of this job step . For among the species of teleost above , a difference was observed in the evolution of the inflammatory response . However , the domain of this technique in the future Denio rerio represent an important tool in the study of mediators and modulators , as well as in understanding and elucidating the pathophysiological mechanisms involved in the acute inflammatory reaction.

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

BELO, MARCO A. A.; OLIVEIRA, MELQUE F.; OLIVEIRA, SUSANA L.; ARACATI, MAYUMI F.; RODRIGUES, LETICIA F.; COSTA, CAMILA C.; CONDE, GABRIEL; GOMES, JULIANA M. M.; PRATA, MARIANA N. L.; BARRA, AYSLAN; et al. Zebrafish as a model to study inflammation: A tool for drug discovery. BIOMEDICINE & PHARMACOTHERAPY, v. 144, DEC 2021. (13/25971-9, 19/19939-1, 18/07098-0)

CHARLIE-SILVA, IVES; FEITOSA, NATALIA MARTINS; MENDONCA GOMES, JULIANA MOREIRA; DE MELO HOYOS, DANIELA CHEMIM; MATTIOLI, CRISTIANO CAMPOS; ETO, SILAS FERNANDES; FERNANDES, DAYANNE CARLA; DE ANDRADE BELO, MARCO ANTONIO; SILVA, JULIANA DE OLIVEIRA; BRANCO DE BARROS, ANDRE LUIS; et al. Potential of mucoadhesive nanocapsules in drug release and toxicology in zebrafish. PLoS One, v. 15, n. 9, SEP 24 2020. (19/19939-1, 13/25971-9, 18/07098-0)

CHARLIE-SILVA, IVES; FEITOSA, NATALIA M.; PONTES, LETICIA G.; FERNANDES, BIANCA H.; NOBREGA, RAFAEL H.; GOMES, JULIANA M. M.; PRATA, MARIANA N. L.; FERRARIS, FAUSTO K.; MELO, DANIELA C.; CONDE, GABRIEL; et al. Plasma proteome responses in zebrafish following lambda-carrageenan-Induced inflammation are mediated by PMN leukocytes and correlate highly with their human counterparts. FRONTIERS IN IMMUNOLOGY, v. 13, p. 21-pg., 2022-09-29. (13/25971-9, 19/19939-1)