On its own, chemical analysis is not enough to derive conclusions on the water quality and/or status, which can be valuably complemented by laboratory bioassays. Single chemical, maximum permissible values, and the sum of pesticide concentrations do not take into account possible patterns of synergism, antagonism, dose level dependencies, or even the dominance of several chemicals within a mixture. In addition, several species from different levels in trophic chains are recommended due to differences in species' sensitivities to chemical compounds that are present.
This study examined the effects of three widely used pesticides that have been previously detected in aquatic systems neighbouring agricultural fields on the early-life stages of the zebrafish Danio rerio. Tests involving single exposures and binary combinations of the s-triazine herbicides (atrazine and terbuthylazine) and the organophosphate insecticide chlorpyrifos were performed. Several endpoints, such as swimming behaviour, morphological abnormalities and mortality, were studied. In addition, the inhibition of acetylcholinesterase (AChE) activity was investigated in order to evaluate the mode of action and toxicity of chlorpyrifos in the presence of these herbicides. Results indicate that both binary mixtures elicited synergistic responses on the swimming behaviour of zebrafish larvae. Moreover, although the herbicides were not effective inhibitors of the AChE on their own, a synergistic inhibition of the enzyme activity was obtained by exposure to mixtures with chlorpyrifos. We observed a correlation between impairment of swimming behaviour of the larvae and inhibition of AChE activity. This study supports previous studies concerning the risk assessment of mixtures since the toxicity may be underestimated when looking only at the single toxicants and not their mixtures.
Previous work showed the existence of ecotoxicity of water samples from the Alqueva reservoir due to the presence of the herbicides atrazine, simazine, terbuthylazine and metolachlor. In the present study we examine the effects of these herbicides singly and as binary mixtures on the growth rate of the microalgae Pseudokirchneriella subcapitata. Usually, the toxicity of mixtures is evaluated in relation to the reference models CA (concentration addiction) and IA (independent action). In this study CA model was selected to evaluate the joint effects of s-triazine herbicides on the growth of algae due to their similar mode of action. Moreover, IA reference model was chosen to evaluate the joint toxicity of the chloroacetanilide metolachlor and the s-triazine herbicides due to their different mode of action. In this study dose ratio was the common deviation obtained on both reference models. In the binary mixtures between atrazine/simazine and atrazine/terbuthylazine the increase of the mixtures toxicity (synergism) was mainly due to atrazine. Also, in the binary mixture between atrazine and metolachlor, atrazine was responsible for the increase (synergism) of the mixture toxicity. In the cases of the binary mixtures between simazine/metolachlor and terbuthylazine/metolachlor, the increase of the mixtures toxicity (synergism) was detected when metolachlor was the herbicide dominant, and antagonism was detected when simazine and terbuthylazine were dominant in both mixtures. This study represents an important step to understand the interactions among herbicides detected previously in the waters of the Alqueva reservoir.
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