Micronuclei and Nuclear Buds Induced by Cyclophosphamide in Crocodylus moreletii as Useful Biomarkers in Aquatic Environments

SIMPLE SUMMARY: Crocodiles are territorial reptiles that are exposed to both aquatic and terrestrial environments. Analyzing their state of health and the environment in which they live is essential to detect changes that could affect them by exposure to agents that damage their genetic material, putting their health or other species that share said habitat, including humans, at risk. There are several ways to determine exposure to harmful agents, a very simple and direct one is to analyze the nuclei of blood cells under a microscope, a test that can also be carried out directly in their habitat and that gives rapid results on the effects of agents on those at that moment they are exposed. This study demonstrates the possibility of analyzing the blood of Crocodylus moreletii to quickly analyze its exposure to toxic agents in a sample of its blood by evaluating two abnormal structures in its cells under the microscope, demonstrating that evidence of damage can be observed only by analyzing a drop of their blood. ABSTRACT: Micronuclei (MN) are used to assess genotoxic exposure, whereas nuclear buds (NBs) have been linked to genotoxic events. Crocodylus moreletii was studied to identify MN and NBs. Three groups were formed: Group 1 (water) and groups 2 and 3 (7 or 10 mg/kg of cyclophosphamide). A drop of blood was obtained daily from the claw tip at 0 to 120 h. Spontaneous micronucleated erythrocytes (MNEs) and erythrocytes with nuclear buds (NBEs) were counted. The frequencies of micronucleated young erythrocytes (MNYEs) and NB young erythrocytes (NBYEs) were evaluated, including the ratio of young erythrocytes (YE)/1000 total erythrocytes. No significant differences were observed in the YE proportion on sampling days; group 1 did not show differences for any parameter, whereas group 2 showed significant differences in MNEs and NBEs, and group 3 showed differences in NBEs and NBYEs. Some mitotic activity in circulation was observed in YEs. In conclusion, NBEs could be a more sensitive biomarker to genotoxic damage than MNEs. The identification of these biomarkers leads us to propose Crocodylus moreletii as a possible environment bioindicator because these parameters could be useful to analyze the in vivo health status of these reptiles and for biomonitoring genotoxic pollutants in their habitats.