Abstract
Simple SummaryCrocodiles 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.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.
Highlights
Aquatic environmental pollution is a serious and growing problem that affects all human activities
No significant differences were observed in the young erythrocytes (YE) proportion values in sampling days
Significant differences were observed in the 7 mg/Kg group in nuclear bud formation in erythrocytes (NBEs) at 120 h (p < 0.03), while in the 10 mg group, significant differences were observed in NBEs at 24 h (p < 0.01), 48 h (p < 0.001), 72 h (p < 0.01), and 96 h (p < 0.03), and in NB young erythrocytes (NBYEs) at 72 h (p < 0.05) and 96 h (p < 0.01)
Summary
Aquatic environmental pollution is a serious and growing problem that affects all human activities. Aquatic ecosystems are regularly the ultimate recipient of many of the pollutants produced by natural and anthropogenic sources [1]. Reliable and practical methods that detect the presence of genotoxic contaminants are scarce [4]. Nuclear protrusions termed “buds” have been described as potential biomarkers of genotoxicity [17,18,19,20]. In leukocytes [17,18] and erythrocytes of parrots [20], nuclear buds (NBs) are markers of genotoxicity, and they can be observed in preparations used to evaluate the presence of micronuclei (MN). In culture of human and pig lymphocytes, the number of NBs increased with the administration of mitomycin-C, so it was proposed that
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