Application of the single-cell gel electrophoresis or comet assay has revolutionized the field of genetic ecotoxicology or eco-genotoxicology. It is a rapid, sensitive and relatively inexpensive method providing the opportunity to study DNA damage (including oxidative damage), repair and cell death (apoptosis) in different cell types without prior knowledge of karyotype and cell turnover rate. The assay has, however, often attracted criticism for its lack of ecotoxicological relevance. In addition, in contrast to genetic toxicology where rapid technical progress has been made to improve cell- and tissue-specific adoption of the assay, only limited advancement has been made to transfer the methodologies to ecotoxicological studies. While reviewing the recent information available in the literature and underscoring the importance of induced genetic damage in natural species, the aims of this article are to (i) highlight and judiciously analyse the ecotoxicological relevance of the assay; (ii) attempt to correlate the comet response with other relevant biological responses or biomarkers; (iii) identify the technical challenges and various factors affecting its application in order to make it reliable, reproducible and robust; (iv) critically compare the technical developments in genetic toxicology and genetic ecotoxicology and (v) evaluate the future developments with respect to applications of the assay. It is suggested that while complementing other ecotoxicological parameters and further improving the methodologies, the comet assay will continue to play an important role in genetic ecotoxicology to determine induced genetic damage, which has significant consequences for short- and long-term survival of the natural or wild species. Information obtained through integrated studies using simultaneous applications of multiple biomarkers on different wild organisms could also provide an holistic dimension of toxicological impact of environmental contaminants for the protection of human health.

/pdf/ecotoxicological-applications-and-significance-of-the-comet-1i32mo0302.pdf

Ecotoxicological applications and significance of the comet assay

This review considers the potential of the Comet assay (or Single Cell Gel Electrophoresis, SCGE) to evaluate the environmental impact of genotoxins in aquatic environments. It focuses on in vivo and in situ studies that have been carried out in various marine and freshwater sentinel species, published in the last 5 years. A large number of the studies reviewed report that the Comet assay is more sensitive when compared with other biomarkers commonly used in genetic ecotoxicology, such as sister chromatid exchanges or micronucleus test. Due to its high sensitivity, the Comet assay is widely influenced by laboratory procedures suggesting that standard protocols are required for both fish and mussel cells. However, there are still a wide variety of personalised Comet procedures evident in the literature reviewed, making comparison between published results often very difficult. Standardization and inter-laboratory calibration of the Comet assay as applied to aquatic species will be required if the Comet assay is to be used routinely by national bodies charged with monitoring water quality.

The Comet assay for the evaluation of genotoxic impact in aquatic environments.

This review discusses several research studies that employed comet assay to evaluate the environmental impact of genotoxins in aquatic environments. It focuses on in vivo and in situ studies of aquatic animals. New chemicals are being added each year to the existing burden of toxic substances in the environment. Excessive agricultural and industrial activities adversely affect biodiversity, threatening the survival of species in a particular habitat, as well as posing disease risks to humans. Some of the chemicals, e.g., pesticides and heavy metals, may be genotoxic to the sentinel species and/or to non-target species, causing deleterious effects in somatic or germ cells. Comet assay is a quick, sensitive, and low-cost technique for detecting DNA strand breakage. However, the comet assay has much more to offer than being an assay for testing DNA strand breaks in animal organs. The use of repair enzymes increases the range of DNA lesions that can be detected with the assay. Comparing data from studies that employed different approaches, such as empirical scoring or comet tail lengths, comet assay is one of the challenging techniques to be utilized in environmental studies. The relative amount of DNA in the comet tail indicates DNA break intensity. The assay has been modified to detect various base alterations by including the digestion of nucleoids with a lesion-specific endonuclease. The determination of DNA damage in these indicator species using the comet test would thus offer information on the genotoxic potential of their habitat at an early stage. This would enable intervention techniques to prevent or mitigate adverse health impacts in sentinel animals and humans.

/pdf/an-overview-of-comet-assay-application-for-detecting-dna-1kh4g5y8.pdf

An Overview of Comet Assay Application for Detecting DNA Damage in Aquatic Animals

Pesticide residues with genotoxic potential reach the aquatic environment and constitute a major issue that give rise to concerns at local, regional, national and global scales. Fish serves as an excellent genetic model for the genetic hazard assessment as they are very sensitive to changes in their environment. As there is a close consortium of DNA damage, mutation and induction of various types of genetic disorders, genotoxicity tests like chromosomal aberration test, micronuclei and comet assay are gaining credence and since past few decades many tests have been developed for evaluating genetic alterations in aquatic organisms. These tests rely on the premise that any change to DNA may have enduring and ardent consequences. Thus, the first aim of the genotoxicology is to describe the outcome produced by toxic substances in various test species, but only from the genetic point of view and to draw conclusions that can be extrapolated to man. This review summarizes the genotoxicity tests developed till date and the role of piscine model in genotoxicology. The text also delves the latest knowledge and thinking on these cardinal approaches for the assessment of aquatic environmental health, management and conservation, besides providing useful repository for the researchers especially dealing with aquatic genotoxicity tests. The perspectives for further research on the use of genotoxicology tests were also highlighted.

https://www.longdom.org/open-access/an-introduction-about-genotoxicology-methods-as-tools-for-monitoring-aquatic-ecosystem-present-status-and-future-perspectives-2150-3508-1000158.pdf

An introduction about Genotoxicology Methods as Tools for Monitoring Aquatic Ecosystem: Present status and Future perspectives

Environmental damages or disturbances to genetic material are studied under the scope of several areas, such as toxicology, molecular biology, biochemistry, and genetics. These chemical or physical injuries on the DNA structure can promote changes and damages that are evaluated by genotoxic studies. Genotoxicity is the harmful action of a xenobiotic agent (chemical or physical) on DNA resulting in genetic mutation. This harm may result in tumors (somatic cells) or be transmitted from generation to generation (germline cells), affecting future generations developing diseases or malformations. The results of damaging activities of these chemical agents range from single and double-strand breaks, cross-linking between DNA bases or between DNA bases and proteins, and formation of DNA adducts when a chemical compound is covalently linked to DNA (Preston and Hoffmann 2001). Contaminants may give rise to chain breaks in four different ways —direct binding of genotoxic compounds; through the effects of oxygen radicals; as a result of interaction with reactive metabolites; and as a consequence of enzymatic action of excision repair mechanism (Eastman and Barry 1992; Speit and Hartmann 1999). The damages described above

/pdf/genotoxicity-and-mutagenicity-2flqo0xt4t.pdf

Genotoxicity and Mutagenicity

Effects of algal extracts (Polysiphonia fucoides) on rainbow trout (Oncorhynchus mykiss): a biomarker approach.

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Effects of algal extracts (Polysiphonia fucoides) on rainbow trout (Oncorhynchus mykiss): a biomarker approach.