Standardization and determination of basal values of Chromosomal Aberrations (CA) and Mitotic Index (MI) on Caiman latirostris (Broad- snouted caiman) lymphocytes as markers of genotoxicity and cytotoxicity

POLETTA GISELA L.; CHACÓN, CF; LÓPEZ GONZÁLEZ EVELYN C.; SIROSKI PABLO A.

Santa Fe

Congreso; 25th Working Meeting of the Crocodile Specialist Group, Santa Fe, Argentina May 7th to 10th 2018.; 2018

PROYECTO YACARE / IUCN-SSC Crocodile Specialist Group (CSG)

Chromosomal Aberrations (CA) and Mitotic Index (MI) are short-term tests that enable the early detection of effects produced by agents that cause chromosomal damage and / or alterations in cell kinetics, giving great information about their mechanisms of action. The aim of this study was to standardize and determine basal values of CA and MI on Caiman latirostris (broad-snouted caiman) lymphocytes, for their use as markers of genotoxicity and cytotoxicity in order to include them into the battery of tests applied routinely in this and other native reptile species.Blood samples were obtained from the spinal vein of three juveniles of C. latirostris and whole blood cultures were made under the optimal conditions previously determined for C. latirostris, to obtain the appropriate lymphocyte stimulation rate and metaphases quality and quantity necessary for the analysis of CA and MI. In order to induce CA, whole blood was exposed in vitro to the known alkylating agent Methyl methanesulfonate (MMS) at 10 and 20 µM, and a negative control (NC) without exposure was used as reference. All treatments were performed in duplicate.With the aim to obtain more information regarding genotoxic effects of MMS, the determination of the frequency of micronuclei (FMN) was included as a complementary biomarker to the induction of CA.The results indicated significant differences in the MI (p