Under laboratory conditions, the effects of thiamethoxam were investigated in larvae, pupae and emerging honey bees after exposure at larval stages with different concentrations in the food (0.00001 ng/μL, 0.001 ng/μL and 1.44 ng/μL). Thiamethoxam reduced the survival of larvae and pupae and consequently decreased the percentage of emerging honey bees. Thiamethoxam induced important physiological disturbances. It increased acetylcholinesterase (AChE) activity at all developmental stages and increased glutathione-S-transferase (GST) and carboxylesterase para (CaEp) activities at the pupal stages. For midgut alkaline phosphatase (ALP), no activity was detected in pupae stages, and no effect was observed in larvae and emerging bees. We assume that the effects of thiamethoxam on the survival, emergence and physiology of honey bees may affect the development of the colony. These results showed that attention should be paid to the exposure to pesticides during the developmental stages of the honey bee. This study represents the first investigation of the effects of thiamethoxam on the development of A. mellifera following larval exposure.

This study sought to evaluate the toxicological effects of graphene oxide (GO) through tests with Danio rerio (zebrafish) embryos, considering the influence of the base washing treatment and the interaction with natural organic matter (i.e., humic acid, HA). A commercial sample of GO was refluxed with NaOH to remove oxidation debris (OD) byproducts, which resulted in a base washed GO sample (bw-GO). This process decreased the total oxygenated groups in bw-GO and its stability in water compared to GO. When tested in the presence of HA, both GO and bw-GO stabilities were enhanced in water. Although the embryo exposure showed no acute toxicity or malformation, the larvae exposed to GO showed a reduction in their overall length and acetylcholinesterase activity. In the presence of HA, GO also inhibited acid phosphatase activity. Our findings indicate a mitigation of material toxicity after OD removal. The difference in the biological effects may be related to the materials’ bioavailability and biophysicochemical interactions. This study reports for the first time the critical influence of OD on the GO material biological reactivity and HA interaction, providing new data for nanomaterial environmental risk assessment and sustainable nanotechnology.

In polluted environments organisms are commonly exposed to a combination of chemicals with different modes of action, and their effects can be additionally modified by natural abiotic conditions. One possible mechanism for interactions in mixtures is via toxicokinetics, as chemicals may alter the uptake, distribution, biotransformation and/or elimination of each other, and all these processes can be affected by temperature. In this study, the effect of temperature (T) on the toxicokinetics of copper (Cu) and chlorpyriphos (CHP), applied either singly or in binary mixtures, was studied in the earthworm Eisenia fetida. The experiments were conducted at 10 or 20 °C and the earthworms were exposed to environmentally realistic concentrations of Cu and/or CHP for 16 d, followed by a depuration period of 4 d in uncontaminated soil. The earthworms were sampled for body Cu and/or CHP concentrations and acetylcholinesterase (AChE) activity measurements. The CHP degradation rate in the soil was substantially higher at 20 °C and in soil treated with Cu. The significant (p < 0.05) inhibition of AChE activity in the earthworms exposed to CHP was found. The effect of Cu was significant only at p < 0.1. No synergistic effect of the parallel CHP and Cu exposure was found. Four days after transferring the earthworms to uncontaminated soil, the AChE activity recovered to the level observed in control animals. The temperature effect on the toxicokinetic parameters was more pronounced for CHP than for Cu. In the case of CHP, the assimilation rate constant (kA) was significantly higher at 20 °C than at 10 °C, both in CHP-only and CHP + Cu treatments. A similar trend was found for the elimination rate constant (kE), but the difference was statistically significant only for non-Cu treatments. In the case of Cu, the general trend of higher kA and kE at 20 °C and in the absence of CHP was observed.

Habitat loss and environmental pollution are among the main causes responsible for worldwide biodiversity loss. The resulting species and population declines affect all vertebrates including reptiles. Especially in industrialized countries, pollution by agrochemicals is of remarkable importance. Here, habitat loss has historically been associated with expansion of agriculture. Species persisting in such environments do not only need to cope with habitat loss, but more recently, also with chemical intensification, namely pesticide exposure. In this study, we examined effects of different fungicide and herbicide applications on the common wall lizard (Podarcis muralis) in grape-growing areas. We used three enzymatic biomarkers (GST, GR, AChE) and for the first time saliva from buccal swabs as a minimal-invasive sampling method for detection. Our results demonstrate absorption of substances by lizards and effects of pesticide exposure on enzymatic activities. Our findings are in accordance with those of previous laboratory studies, although samples were retrieved from natural habitats. We conclude that buccal swabs could become a useful tool for the detection of pesticide exposure in reptiles and have the potential to replace more invasive methods, such as organ extraction or cardiac puncture. This is an important finding, as reptiles are non-target organisms of pesticide applications, and there is a strong need to integrate them into pesticide risk assessments.

Microcystin-LR (MCLR) has been reported to cause developmental neurotoxicity in zebrafish, but there are few studies on the mechanisms of MCLR-induced transgenerational effects of developmental neurotoxicity. In this study, zebrafish were exposed to 0, 1, 5, and 25 μg/L MCLR for 60 days. The F1 zebrafish embryos from the above-mentioned parents were collected and incubated in clean water for 120 h for hatching. After examining the parental zebrafish and F1 embryos, MCLR was detected in the gonad of adults and F1 embryos, indicating MCLR could potentially be transferred from parents to offspring. The larvae also showed a serious hypoactivity. The contents of dopamine, dihydroxyphenylacetic acid (DOPAC), serotonin, gamma-aminobutyric acid (GABA) and acetylcholine (ACh) were further detected, but only the first three neurotransmitters showed significant reduction in the 5 and 25 μg/L MCLR parental exposure groups. In addition, the acetylcholinesterase (AChE) activity was remarkably decreased in MCLR parental exposure groups, while the expression levels of manf, bdnf, ache, htr1ab, htr1b, htr2a, htr1aa, htr5a, DAT, TH1 and TH2 genes coincided with the decreased content of neurotransmitters (dopamine, DOPAC and serotonin) and the activity of AChE. Neuronal development related genes, α1-tubulin, syn2a, mbp, gfap, elavl3, shha and gap43 were also measured, but gap43 was the gene only up-regulated. Our results demonstrated MCLR could be transferred to offspring, and subsequently induce developmental neurotoxicity in F1 zebrafish larvae by disturbing the neurotransmitter systems and neuronal development.

In this study, we demonstrated an assay with turn-on fluorescence for monitoring cerebrospinal acetylcholinesterase (AChE) fluctuation as a biomarker for organophosphorus pesticides (OPs) poisoning and management based on single layer MnO2 nanosheets with graphene quantum dots (GQDs) as signal readout. Initially, the fluorescence of GQDs was quenched by MnO2 nanosheets mainly due to the inner filter effect (IFE). However, with the presence of reductive thiocholine (TCh), the enzymatic product, hydrolyzed from acetylthiocholine (ATCh) by AChE, the redox reaction between MnO2 and TCh occurred, leading to the destruction of the MnO2 nanosheets, and thereby IFE was diminished gradually. As a consequence, the turn-on fluorescence of GQDs with the changes in the spectrum of the dispersion constituted a new mechanism for sensing of cerebrospinal AChE. With the method developed here, we could monitor cerebrospinal AChE fluctuation of rats exposed to OPs before and after therapy, and could thereby open up the pathway to a new sensing platform for better understanding the mechanism of brain dysfunctions associate with OPs poisoning.

This field study is intended to propose a global methodology to assess and monitor the water quality of the gulf of Morbihan, a littoral ecosystem under increasing anthropic pressure. To this end, the Locmariaquer site, where Crassostrea gigas is extensively cultivated, was selected to perform a one-year follow-up of tissular glutathione S-transferase and acetylcholinesterase specific activities in this filter feeder organism. Calculation of an integrated index, corresponding to the ratio of the two enzymes activities, allowed to discriminate from the environmental noise, several clusters which could be representative environmental stress, potentially latent pollution. Moreover, the estrogenic activity was assessed in water samples collected at Locmariaquer and other strategic sites of the gulf. The results evidenced a low estrogenic-disrupting compound contamination of waters. Overall, this methodology produced an accurate outlook of a basal state for the gulf and could be developed in the context of a chronic monitoring of this site.

Amphibians are the most threatened vertebrate group with a third of currently known species endangered with extinction, as a result of climate change, habitat loss, disease-introduced exotic species, and pollution. Because of their vulnerability, they have often been used as environmental quality indicators, as well as laboratory models for toxicological research. Given the sensitivity of amphibians to changes in their surrounding environment, including pollution, it was deemed important to define a non-lethal technique based on the evaluation of a set of biomarkers in different tissues of neotenic individuals of Ambystoma velasci. The levels of acetylcholinesterase (AChE), butyrylcholinesterase (BChE), carboxylesterase (CaE), alkaline and acid phosphatases (ALP, ACP), glutathione s-transferase (GST), 7-ethoxyresorufin-O-deethylase (EROD), and superoxide dismutase (SOD) activities, as well as the oxygen radical absorption capacity (ORAC) were measured in tail, gills, liver, plasma, and brain samples. Significant tissue-specific differences were observed for all biomarkers with the exception of ACP. The highest values of specific activity for most biomarkers were detected in the liver. However, the levels measured in gills were very close to those observed in the liver and showed fewer variations than other tissues. These findings suggest that the sampling of gills could be used to evaluate pollution biomarkers in salamanders without apparent harm, as this tissue quickly regenerates.

The polychlorinated dibenzo-p-dioxins/dibenzofurans (PCDD/Fs) are persistent environmental pollutants. Recently, there have been an increasing demand to assess different biomarkers as early alarming indicators of environmental pollution. This study is the first to investigate the effects of PCDD/Fs on acetylcholinesterase (AChE) activity and histopathology of the body wall (epidermis, circular, and longitudinal muscles) of earthworm Eisenia andrei (Oligochaeta, Lumbricidae) using acute filter paper toxicity test. It is also exploring the selected biomarkers as a potential tool for evaluating soil quality. Earthworms were exposed to 0, 0.5, 1.0, and 1.5 ng/cm² PCDD/Fs. The treated worms expressed progressive abnormal morphological signs with incrementing doses of dioxins, such as coiling, curling, and body swelling. At the highest dose, some worms demonstrated skin discoloration, loss of body segmentation, and body part detachment. The AChE activity was significantly decreased (p < 0.001) in all treated animals compared to control. The mixture induced circular muscle hyperplasia at 0.5 ng/cm². However, the mixture at 1.5 ng/cm² caused epidermal atrophy with cell pyknosis and necrosis in all layers of the body wall. Image analysis revealed significant reduction in the thickness of the epidermis (p < 0.001) at all doses with relative to control. Herein, we report that 48 h of acute exposure of E. andrei to dioxins/furans induced morphological changes, reduced the activity of AChE, and induced histopathological alterations. The outcomes can be utilized as endpoints that could be added to earthworm’s standardized short tests for ecotoxicity studies.

Little is known about the potential toxicity of Cu nanoparticles (nCu), Cr nanoparticles (nCr), and their mixtures to aquatic organisms. To fill this gap, a comprehensive toxicity assessment was conducted using Daphnia magna as a test organism, including a 48-h acute toxicity test, a 21-day chronic test, and a feeding experiment. Four biomarkers were estimated after exposure to nCu, nCr, and their mixtures for 7 days, including acetylcholinesterase (AChE), catalase (CAT), superoxide dismutase (SOD), and glutathione S-transferase (GST). The results at all endpoints showed that nCu was more toxic than nCr. The 48-h median lethal concentration values of nCu and nCr were 0.63 and 1.57 mg/L, respectively. Significant inhibition of reproduction and growth of D. magna was found, and the intrinsic rate of natural increase was a sensitive parameter for nCu and nCr during the 21-day exposure. A concentration-dependent decrease in filtration and ingestion was observed which was consistent with inhibition of reproduction and growth of D. magna. The biochemical responses revealed an increase in GST activity and decrease in AChE activity, while SOD and CAT activities were increased at low concentrations and decreased at high concentrations for all exposures. Collectively, our results confirmed that nanoscale Cu and Cr can exert negative effects at different levels on D. magna.