A general report on the use of the Allium test as cytotoxicological and genotoxicological assay is proposed, with particular emphasis about the standardization of the test in several common applications. The intraspecific variation in Allium cepa has been overlooked, as in most investigations no mention is made about origin and denomination of the onion cultivar used. A standardization of the used material would allow a better generalization of the results, since we cannot be sure that all cultivars would give the same response. A more frequent use of transmission electron microscopy (TEM) investigation is proposed. Even if relatively time consuming and not available in all laboratories, it may help to better understand the mechanism of cytotoxicity, since many morphological characters may appear similar but be arisen from different processes observable only with TEM. About statistical testing, tests other than chi-squared may be used in case of a lower amount of data. The most commonly used statistical tests are the parametric tests ANOVA and Student's t, and the nonparametric tests Kruskal-Wallis and Mann-Whitney U, for analysis of variance. Tests should be used also to assess the minimal sample dimension for obtaining significance, since data collection (microscope observation) appears to be one of the main bottle necks of the test. Also the use of the Allium test for testing liposomes and other nanovectors for drug delivery is proposed, in order to assess the cytotoxicity of these types of medium and the possible increase in cytotoxicity of the associated drug.
For the first time a study was conducted of the effects of the change in the concentration of sodium chloride in water on the biomarkers of oxidative stress (enzymes: catalase (CAT), glutathione reductase (GR), glutathione-S-transferase (GST), and the levels of reduced glutathione (GSH), and a marker of peroxidation lipids (LPO) – malonic dialdehyde (MDA)) in the gills of the freshwater mollusc Anodonta cygnea. Adult specimens of A. cygnea of both sexes with a shell length of 70–100 mm were selected for the study, a total of 36 individuals. The molluscs were collected in one of the bays on the southwestern coast of the Volga reaches of the Rybinsk Reservoir (reservoir of the cascade of the upper Volga). The molluscs were kept in the laboratory in aquariums with river water and sand collected from the place of the molluscs' collection. During the experiment, a solution of NaCl 3 g/l in river water was introduced into the container with molluscs. Samples of molluscs’ tissues were taken before application of the saline solution, 40 and 120 min after changing the salt concentration to 3 g/l, and 20 and 60 min after changing the NaCl solution to river water. The change in the concentration of NaCl in water (0–3 g/l) did not affect the content of water-soluble protein in the gills of molluscs during short-term exposure. The change in NaCl concentration in water caused the intensification of LPO processes in the gills of A. cygnea. The change in the activity of the enzymes of the antioxidant system and the increase in the concentration of GSH in A. cygnea gills, indicates their involvement in protecting freshwater molluscs from NaCl-induced oxidative stress. The change in the functioning of the glutathione system may be a compensatory mechanism for changing the NaCl concentration in water. Such biomarkers of oxidative stress as malonic dialdehyde and enzymes of the antioxidant system are sensitive indicators of changes in NaCl concentration in water in freshwater bivalve molluscs, for example A. cygnea.
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