Cytotoxic and estrogenic activity of chlorpyrifos and its metabolite 3,5,6-trichloro-2-pyridinol. Study of marine yeasts as potential toxicity indicators.

Chlorpyrifos (CP) is one of the organophosphate insecticides most used worldwide today. Although the main target organ for CP is the nervous system triggering predominantly neurotoxic effects, it has suggested other mechanisms of action as cytotoxicity and endocrine disruption. The risk posed by the pesticide metabolites on non-target organisms is increasingly recognized by regulatory agencies and natural resource managers. In the present study, cytotoxicity and estrogenic activity of CP, and its principal metabolite 3,5,6-trichloro-2-pyridinol (TCP) have been evaluated by in vitro assays, using two mammalian cell lines (HEK293 and N2a), and a recombinant yeast. Results indicate that TCP is more toxic than CP for the two cell lines assayed, being N2a cells more sensitive to both compounds. Both compounds show a similar estrogenic activity being between 2500 and 3000 times less estrogenic than 17ß-estradiol. In order to find new toxicity measurement models, yeasts isolated from marine sediments containing CP residues have been tested against CP and TCP by cell viability assay. Of the 12 yeast strains tested, 6 of them showed certain sensitivity, and a concentration-dependent response to the tested compounds, so they could be considered as future models for toxicity tests, although further investigations and proves are necessary.

Acute toxicity of chlorpyrifos and its metabolite 3,5,6-trichloro-2-pyridinol alone and in combination using a battery of bioassays.

Acute toxicity of chlorpyrifos (CP) and its principal metabolite 3,5,6-trichloro-2-pyridinol (TCP) alone and in combination have been evaluated using a test battery comprising aquatic organisms from different trophic levels: luminescent marine bacteria Aliivibrio fischeri, freshwater unicellular alga Pseudokirchneriella subcapitata, and cladoceran Daphnia magna. As expected, D. magna was the more sensitive organism to the compounds tested, being CP more toxic than its metabolite. On the contrary, TCP was found to be more toxic than its parental compound to A. fischeri and P. subcapitata. In all cases, the mixture of CP and its metabolite was more toxic than the compounds tested separately, multiplying between 5 and 200 times CP toxicity level and up to 15 times TCP toxicity level. These results indicate that the co-existence of parent chemical and its degradation product in the environment can result in a synergic interaction involving high risk to the aquatic ecosystems. Graphical abstract.

Ecotoxicological assessment of perchlorate using in vitro and in vivo assays.

Perchlorate is an inorganic ion widespread in the environment, generated as a natural and anthropogenic pollutant, with known endocrine disruption properties in the thyroid gland. Nonetheless, there are few reports of its ecotoxicological impact on wildlife. The aim of this study was to evaluate the adverse effects of KClO4 exposure on different cell lines, HEK, N2a, and 3T3, as well as in ecological models such as Vibrio fischeri, Pseudokirchneriella subcapitata, Daphnia magna, and Eisenia fetida. Perchlorate exhibited similar toxicity against tested cell lines, with LC50 values of 19, 15, and 19 mM for HEK, N2a, and 3T3, respectively; whereas in V. fischeri, the toxicity, examined as bioluminescence reduction, was considerably lower (EC50 = 715 mM). The survival of the freshwater algae P. subcapitata was significatively impaired by perchlorate (LC50 = 72 mM), and its effect on the lethality in the crustacean D. magna was prominent (LC50 = 5 mM). For the earthworm E. fetida, the LC50 was 56 mM in soil. In this organism, perchlorate induced avoidance behavior, weight loss, and decreased egg production and hatchling, as well as morphological and histopathological effects, such as malformations, dwarfism, and necrosis. In conclusion, perchlorate toxicity varies according to the species, although E. fetida is a sensitive model to generate information regarding the toxicological impact of KClO4 on biota.