Comet assay protocol for Bombus atratus fat body and pericardial cells (Hymenoptera, bombini) at a safe concentration of mercury
| Autor: | Letícia Ceschi-Bertoli, Fábio Camargo Abdalla, Paulo José Balsamo, Felipe Augusto Pinto Vidal |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Environmental Engineering
Health Toxicology and Mutagenesis Fat Body 0208 environmental biotechnology Population chemistry.chemical_element 02 engineering and technology 010501 environmental sciences medicine.disease_cause 01 natural sciences chemistry.chemical_compound medicine Animals Environmental Chemistry Ecotoxicology Bioassay education Cells Cultured 0105 earth and related environmental sciences education.field_of_study Chromatography Bombus atratus Acridine orange Public Health Environmental and Occupational Health Mercury General Medicine General Chemistry Bees Pollution 020801 environmental engineering Mercury (element) Comet assay chemistry Environmental Pollutants Comet Assay Pericardium Brazil Genotoxicity DNA Damage |
| Zdroj: | Chemosphere. 261:127752 |
| ISSN: | 0045-6535 |
| DOI: | 10.1016/j.chemosphere.2020.127752 |
| Popis: | The decline of the Bombus population is closely related to the presence of environmental pollutants. Among these pollutants, trace metals represent a major concern, which includes mercury, a known genotoxic substance. The induction of genotoxicity may be demonstrated by the comet assay (a.k.a. single-cell gel electrophoresis), a simple and sensitive method for DNA damage estimating. The current work provided, for the first time, a protocol of comet assay for Bombus atratus using mercury as a standard chemical at safe concentrations according to the Environment National Council of Brazil, and the World Health Organization. Bees were collected and divided into three groups (n = 11 each), in which the exposed groups received a 0.2 ppb or a 1 ppb of mercury solution, and the control group received water. The bioassay was performed for 48 h at controlled temperature and humidity conditions, according to the OECD guideline toxicological test method for B. terrestris. The samples were stained with different dyes to observe the efficacy of each one. Variations of parameters in methodology, such as concentration and time of exposure to lysis solution as well as the electrophoretic process, allowed the observation of comets at different levels. DAPI and acridine orange presented an unstable fluorescence, and silver nitrate dye was more effective. Therefore, the comet assay was shown to be an effective method to evaluate genotoxic effects in bees. The obtained results may be helpful for the establishment of a suitable protocol for future genotoxicity assessment in neotropical bees using different doses of xenobiotics. |
| Databáze: | OpenAIRE |
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