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 non-parametric 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.

Silver nanoparticles are one of the most commonly used nanomaterials. However, there remains insufficient information on their genotoxic effects. The goal of this study was to investigate cytotoxic and genotoxic effects of chitosan-coated AgNPs using the well-established Allium test. Root tip cells were treated with solutions at different concentrations (1, 2.5, 5 and 50 mg l⁻¹) of chitosan-coated AgNP (size: 10–30 nm; organic coat: 2–5 nm). On the same slide mitotic abnormalities, chromosome aberrations and micronuclei were detected. Also mitotic and phase indexes were analyzed. No cytotoxic or genotoxic effects were found at concentrations below 5 mg l⁻¹. The absence of induction of chromosomal and mitotic abnormalities by chitosan-capped AgNPs at low concentrations is possibly due to the capping, which may partly protect the cells from direct interaction with the AgNPs. Mitotic and chromosomal abnormalities and micronuclei were detected at a concentration of 50 mg l⁻¹. Significant increase in mitotic index was found at 5 and 50 mg l⁻¹ concentrations of AgNP. The data demonstrated that chitosan-coated AgNP exhibit both clastogenic and aneugenic activity.

Heracleum sosnowskyi is one of the most aggressive alien species in Europe. However, no information is available on its genotoxic effects. The goal of this study was to investigate cytological aberrations and genotoxic effects of H. sosnowskyi juice using the well-established Allium test. Root tip cells were treated with varying concentrations (1, 5, 10, 50, 100 and 300 ml l^–1) of solution in water of H. sosnowskyi juice. The same slides with root tips were used for mitotic abnormalities, chromosome aberrations and micronuclei scoring. Mitotic and phase indexes were also analyzed. No genotoxic effects were found at a concentration of 1 ml l^–1. Mitotic abnormalities were detected at concentrations of 10, 50, 100, and 300 ml l^–1. Chromosomal abnormalities and micronuclei were detected at concentrations of 50, 100, and 300 ml l^–1. Significant decrease in mitotic index was found at 5, 10, 50, 100, and 300 ml l^–1 concentrations of H. sosnowskyi juice. Lagging chromosomes were the most common abnormalities. The data demonstrated that H. sosnowskyi juice exhibits both clastogenic and aneugenic activity. However, aneugenic activity is more pronounced.

Biomarkers of oxidative stress such as catalase (CAT), glutathione S-transferase (GST), glutathione reductase (GR) activity, and malondialdehyde and reduced glutathione content, as well as heavy metal concentrations (HM: Pb, V, Cr, Mn, Co, Ni, Cu, Zn, and Cd), were studied in Dreissena polymorpha tissues. Mussels were collected on three sites located on the Rybinskoe Reservoir different in levels of anthropogenic pressure: the most polluted sites were 1 and 2 and site 3 was relatively clean. Mussels from sites 1 and 2 had higher concentrations of HM (Pb, V, Cr, Mn, Ni, Cu, and Mn) and their response to the pollutant action was manifested in increased processes of lipid peroxidation (LPO), the activation of CAT, and elevated level of GHS.

Cyanobacteria, which develop abundantly in aquatic ecosystems, can be harmful to humans and animals not only by releasing toxins that cause poisoning but also by provoking cytogenetic effects. The influence of the mass development of cyanobacteria on the genotoxic properties of natural water has been studied in model ecosystems (microcosms) with different compositions of biotic components (zooplankton, amphipods and fish). The validated plant test system “Allium test” was used in this study. Genotoxic effects were detected at microcystin concentrations below those established by the World Health Organization (WHO) for drinking water. In all experimental treatments, cells with disorders such as polyploidy and mitotic abnormalities associated with damage to the mitotic spindle, including c-mitosis, as well as lagging chromosomes were found. Genotoxic effects were associated with the abundance of cyanobacteria, which, in turn, depended on the composition of aquatic organisms in the experimental ecosystem. Fish, to a greater extent than other aquatic animals, maintain an abundance of cyanobacteria. After one month, in microcosms with fish, mitotic abnormalities and polyploidy continued to be detected, whereas in other treatments, there were no statistically significant genotoxic effects. In microcosms with amphipods, the number and biomass of cyanobacteria decreased to the greatest extent, and only one parameter of genotoxic activity (frequency of polyploidy) significantly differed from the control.