Imidacloprid is a widely used pesticide that belongs to the class of neonicotinoids. There is a piece of rising evidence that neonicotinoids exert cytotoxic effects in non-target organisms including vertebrate species such as mammals. Nevertheless, dose-limiting toxicity and molecular mechanisms of neonicotinoids' deleterious effects are still poorly understood. In accord to imidacloprid fate in the environment, the most of used pesticide is absorbed in the soil. Therefore, earthworms, which are prevailing soil organisms, could be considered as a target of neonicotinoids toxicity. The earthworm’s simple nervous system is a prospective model for neurotoxicological studies. We exposed earthworms to imidacloprid in a paper contact test with a doses range of 0.1‑0.4 µg/cm2 for 14 days. In the present work, we studied the imidacloprid effect on oxidative stress generation and neuronal marker neuron-specific enolase (NSE) expression. The exposure to imidacloprid induced a dose-dependent decrease in NSE. Both reactive oxygen species production and lipid peroxidation level were upregulated as well. Observed NSE decline suggests imidacloprid-caused disturbance in earthworm neuron cells. Obtained data have shown that relatively low doses of imidacloprid are potent to induce cytotoxicity in neurons. Furthermore, neurotoxicity could be recognized as one of an individual scenario of the general imidacloprid toxicity. Thus, presented results suggest the cytotoxicity of imidacloprid low doses in non-target organisms and hypothesize that NSE downregulation could be estimated as a biomarker of neonicotinoid cytotoxicity in a nervous system of non-insect species.
Currently, most amphibian populations in the world exist under the influence of numerous stress factors. Among them, the main factors that affect almost all terrestrial animals, namely, the fragmentation of habitats, environmental pollution and anthropic transformation of landscapes. Moreover, those factors are joined by negative causes that affect only amphibians – specific viral and fungal infections: ranaviruses (Ambystoma tigrinum virus (ATV), Bohle iridovirus (BIV), and frog virus 3) (also dangerous for some reptiles) and chytrid fungi (Batrachochytrium dendrobatidis and B. salamandrivorans). All these factors are one of the main reasons for the current global decline of amphibian populations in the world. In today's world, agricultural chemicals are one of the most important in terms of toxicity, environmental emissions and total area of impact. Among them, a significant proportion are formed by synthetic insecticides, which include pyrethroids and neonicotinoids. Pollution by these substances has a negative impact on amphibian populations, despite the relatively short period of their life in the environment. The vast majority of studies on the effects of pyrethroid and neonicotinoid insecticides were made in laboratory experiments with the larval stages of tailless amphibians. Tadpoles are easy to get in sufficient quantities and kept in the laboratory. Cypermethrin reduces the viability of tadpoles and causes precocious metamorphosis of survived larvae. However, in a combination with other pesticides, it delays metamorphosis. In addition, embryos were more resistant to pyrethroids than tadpoles. Pyrethroid pesticides cause spasms indicating adverse neurological effects. Formation of oral apparatus abnormalities in tadpoles, anisochromasia and increasing number of immature erythrocytes are also caused by pyrethroids. Neonicotinoids show similar effects. Under the action of imidacloprid the erythrocytes are also disturbed that is showed up in the DNA damage and micronuclei formation. Neurological disorders are manifested in the ability to perceive or respond to a predator, disorientation, erratic movement and loss of balance. Pyrethroids and neonicotinoids have been shown cause a variety of disorders: increase mortality and reduce survival of tadpoles; have a teratogenic effect and affect the metamorphosis and morphological parameters of amphibians; change many biochemical parameters that characterize the protein metabolism and oxidative stress; have genotoxic effects and affect the state of the nervous system and animal behaviour. Most of these parameters are proposed for use as biomarkers of pesticide intoxication.
Imidacloprid is one of the most widely used insecticides in the world. The neurotoxicity of imidacloprid in adult amphibians has not been studied thoroughly. We investigated the expression of glial fibrillary acidic protein (GFAP), neurofilament light chain (NfL) and angiostatin in the amphibian brain to identify valid biomarkers of low dose imidacloprid exposure. For the experiment, 30 individuals of the marsh frog Pelophylax ridibundus were selected. The amphibians were divided into five groups. The duration of the experiment was 7 and 21 days. The exposure concentrations were 10 and 100 µg/L. The results of the study revealed a decrease in the expression of GFAP after 7 days in the exposure groups of 10 and 100 μg/L. An increase in the level of NfL was observed in the group exposed to 10 μg/L after 21 days of the experiment. The angiostatin level was increased after 7 days at 10 µg/L and after 21 days at 100 µg/L. The data obtained indicate that low concentrations of imidacloprid can cause neurotoxic effects in the brain of P. ridibundus. Such effects can have a significant impact on amphibian populations. According to the results of the study of the expression level of GFAP, NfL and angiostatin, it can be stated that imidacloprid has a neurotoxic effect on adult marsh frogs. The studied indicators can be promising biomarkers of environmental pollution by neonicotinoids.
Imidacloprid is a widely used pesticide that belongs to the class of neonicotinoids. There is a piece of rising evidence that neonicotinoids exert cytotoxic effects in non-target organisms including vertebrate species such as mammals. Nevertheless, dose-limiting toxicity and molecular mechanisms of neonicotinoids' deleterious effects are still poorly understood. In accord to imidacloprid fate in the environment, the most of used pesticide is absorbed in the soil. Therefore, earthworms, which are prevailing soil organisms, could be considered as a target of neonicotinoids toxicity. The earthworm’s simple nervous system is a prospective model for neurotoxicological studies. We exposed earthworms to imidacloprid in a paper contact test with a doses range of 0.1‑0.4 µg/cm2 for 14 days. In the present work, we studied the imidacloprid effect on oxidative stress generation and neuronal marker neuron-specific enolase (NSE) expression. The exposure to imidacloprid induced a dose-dependent decrease in NSE. Both reactive oxygen species production and lipid peroxidation level were upregulated as well. Observed NSE decline suggests imidacloprid-caused disturbance in earthworm neuron cells. Obtained data have shown that relatively low doses of imidacloprid are potent to induce cytotoxicity in neurons. Furthermore, neurotoxicity could be recognized as one of an individual scenario of the general imidacloprid toxicity. Thus, presented results suggest the cytotoxicity of imidacloprid low doses in non-target organisms and hypothesize that NSE downregulation could be estimated as a biomarker of neonicotinoid cytotoxicity in a nervous system of non-insect species.
Agricultural activity in the global world is accompanied by the use of a significant number of synthetic insecticides for the control of insect pests. Pyrethroid and neonicotinoid insecticides are among the widely used insecticides in many countries for the control of crop pests. They are a generation of synthetic insecticides that have replaced the more environmentally stable organophosphorus and organochlorine compounds. Pyrethroid and neonicotinoid insecticides were thought to have low toxicity to vertebrates, leading to their widespread use and increased production. However, many studies have emerged in recent decades that have shown that, under certain conditions, these substances can cause significant damage to the internal systems of amphibians. Recently, special studies have also revealed the toxic effects of pyrethroids and neonicotinoids on the post-metamorphic stages of amphibians, which had previously been ignored. It has also been noted that abnormalities in gastrointestinal tract functions occur, leading to abnormalities in the digestive system. Pyrethroid and neonicotinoid insecticides have been shown to affect the biochemical and histological parameters of amphibians. The possible genotoxicity of these insecticides resulted in producing erythrocytes with abnormal nuclei and an increased number of micronuclei in amphibian cells. Meanwhile, changes in the activity of antioxidant enzymes and increases in lipid peroxidation products could be used as biomarkers of oxidative stress in amphibians under the influence of pyrethroid and neonicotinoid insecticides. The available literature also indicates that these insecticides appear to affect the nervous system of amphibians and induce changes in their behaviour. At the same time, our data suggest that it is neuromolecular biomarkers that can be practised to determine the toxic effects of insecticides on non-target species. Such biomarkers can be used in the context of the low-dose influence of insecticides, which however requires additional research on amphibians.
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