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 ofthe honey bee. This study represents the first investigation of the effects of thiamethoxam on the development of A. mellifera following larval exposure.

Understanding the metabolic defense and compensation to maintain homeostasis is crucial for assessing the potential health risk of organic pollutants in crops. Currently, limited understanding is available regarding the targeted metabolic pathways and response mechanism under contaminant stress. This study showed that ciprofloxacin (CIP) at the environmental concentrations (1, 5, 25, 50 mg/L) did not significantly inhibit growth or cause severe oxidative damage to rice (Oryza sativa L.). Instead, the increment in CIP concentration induced a series of sequential metabolic disorders, which were characterized predominantly by primary and secondary metabolic disturbances, including phenylpropanoid biosynthesis, the carbohydrate, lipid and amino acid metabolism. After CIP in vivo exceeded a certain threshold level (>0.29 mg/g dry weight), β-glucosidases (BGLUs) mediated the transition from the activation of the genes related to phenylpropanoid biosynthesis to the inhibition of the genes related to carbohydrate metabolism in rice. In particular, starch and sucrose metabolism showed the most profound perturbation stressed by environmental concentrations of CIP (5 mg/L) and other tested organic pollutants (10 μg/L of tricyclazole, thiamethoxam, polybrominated diphenyl ethers, and polychlorinated biphenyls). Besides, the key genes encoding endoglucanase and BGLU were significantly downregulated (|log₂FC| > 3.0) under 100 μg/L of other tested organic pollutants, supporting the transition from the activation of secondary defense metabolism to the disruption of primary energy metabolism. Thus, in addition to bioaccumulation, changes in BGLU activity and starch and sucrose metabolism can reflect the potential adverse effects of pollutants on rice. This study explained the stepwise metabolic and transcriptional responses of rice to organic pollutants, which provided a new reference for the comprehensive evaluation of their environmental risks.

Neonicotinoid pesticide use is widespread and highly debated, as evidenced by recent attention received from the public, academics and pesticide regulatory agencies. However, relatively little is known about the physiological effects of neonicotinoid insecticides on aquatic vertebrates. Amphibians (larval stages in particular) are excellent vertebrate bioindicators in aquatic systems due to their risk of exposure and sensitivity to environmental stressors. Previous work with wood frog (Rana sylvatica) tadpoles exposed to formulated products containing thiamethoxam or clothianidin in outdoor mesocosms found significant shifts in leukocyte profiles, suggesting the tadpoles were physiologically stressed. The main objective of the present study was to characterize this stress response further using complementary measures of stress after exposure to clothianidin on northern leopard frogs (Rana pipiens) during their aquatic larval stages. Laboratory static-renewal exposures were conducted over eight weeks with the technical product clothianidin at 0, 0.23, 1, 10 and 100 μg/L, and diquat dibromide at 532 μg/L was used as a positive control. We assessed tadpole leukocyte profiles and measures of oxidative stress as these sub-lethal alterations could affect amphibian fitness. We found changes in several types of leukocytes at 1 and 10 μg/L, suggesting that these tadpoles exhibited signs of mild physiological stress. Clothianidin also induced an oxidative stress response at 0.23, 1 and 100 μg/L. However, we found no differences in survival, growth, development time or hepatosomatic index in frogs exposed to clothianidin. Our study indicates that tadpoles chronically exposed to clothianidin have increased stress responses, but in the absence of concentration-response relationships and effects on whole-organism endpoints, the implications on the overall health and fitness of these changes are unclear.

Aquatic pollution from emerging organic contaminants (EOCs) is of key environmental importance in India and globally, particularly due to concerns of antimicrobial resistance, ecotoxicity and drinking water supply vulnerability. Here, using a broad screening approach, we characterize the composition and distribution of EOCs in groundwater in the Gangetic Plain around Patna (Bihar), as an exemplar of a rapidly developing urban area in northern India. A total of 73 EOCs were detected in 51 samples, typically at ng.L⁻¹ to low μg.L⁻¹ concentrations, relating to medical and veterinary, agrochemical, industrial and lifestyle usage. Concentrations were often dominated by the lifestyle chemical and artificial sweetener sucralose. Seventeen identified EOCs are flagged as priority compounds by the European Commission, World Health Organisation and/or World Organisation for Animal Health: namely, herbicides diuron and atrazine; insecticides imidacloprid, thiamethoxam, clothianidin and acetamiprid; the surfactant perfluorooctane sulfonate (and related perfluorobutane sulfonate, perfluorohexane sulfonate, perfluorooctanoic acid and perfluoropentane sulfonate); and medical/veterinary compounds sulfamethoxazole, sulfanilamide, dapson, sulfathiazole, sulfamethazine and diclofenac. The spatial distribution of EOCs varies widely, with concentrations declining with depth, consistent with a strong dominant vertical flow control. Groundwater EOC concentrations in Patna were found to peak within ∼10 km distance from the River Ganges, indicating mainly urban inputs with some local pollution hotspots. A heterogeneous relationship between EOCs and population density likely reflects confounding factors including varying input types and controls (e.g. spatial, temporal), wastewater treatment infrastructure and groundwater abstraction. Strong seasonal agreement in EOC concentrations was observed. Co-existence of limited transformation products with associated parent compounds indicate active microbial degradation processes. This study characterizes key controls on the distribution of groundwater EOCs across the urban to rural transition near Patna, as a rapidly developing Indian city, and contributes to the wider understanding of the vulnerability of shallow groundwater to surface-derived contamination in similar environments.

Neonicotinoids (NEOs) have become the most widely used insecticides in the world. However, information on the NEO concentrations in tap water based on nationwide surveys is limited in China. In this study, the levels of six NEOs were measured in tap water samples collected from 38 cities in China. Across all sampling locations, the overall frequency of detection for at least one NEO was 100%, indicating that NEOs are ubiquitous in tap water from China. Imidacloprid was the most abundant NEO (median: 7.59 ng/L), followed by (in decreasing order) clothianidin (5.46 ng/L), dinotefuran (4.55 ng/L), thiamethoxam (4.50 ng/L), acetamiprid (2.72 ng/L), and thiacloprid (0.38 ng/L). Significantly positive correlations (r = 0.655–0.902, p < 0.001) among all pairs of NEOs were observed, which showed that the sources of NEOs in tap water were common or related. Across all sampling locations, regional differences in concentrations (ΣNEOs; sum of six NEOs varied from 0.79 ng/L to 415 ng/L) were observed within China, showing an increasing trend from northern to southern China. Although the estimated daily intake of NEOs were much lower than the reference dose, the potential health risk of NEOs via tap water consumption should raise more public concern considering the high detection rates of NEOs in tap water.

It is necessary to understand the interactions between different pesticides in ecotoxicology because pesticides never appear as individual compounds but rather in combinations with other compounds. In this study, we planned to explicate the combined toxic effect of myclobutanil (MYC) and thiamethoxam (THI) on the zebrafish (Danio rerio) by adopting multiple biomarkers. Results unraveled that the 96-h LC₅₀ values of MYC to D. rerio at various life phases ranged from 5.2 to 10.3 mg L⁻¹, which were lower than those of THI ranging from 147 to 246 mg L⁻¹. Combinations of MYC and THI exhibited synergetic toxicity to zebrafish embryos. The activities of antioxidative enzymes (T-SOD, Cu/Zn-SOD and POD) and detoxification enzyme (GST) were obviously varied in most of the MYC, THI and combined exposures compared to the control. The mRNA expressions of eight genes (Cu-sod, cas3, il-8, cxcl, erα, crh, cyp17 and dio1) involved in antioxidation, apoptosis, immunity and endocrine were obviously altered in the combined exposure of MYC and THI compared to their individual exposures. Our findings hinted the threats when YMC and THI co-existed, which would be beneficial for the risk assessments of pesticide mixtures.

Production of chrysanthemum (Dendranthema grandiflora) in greenhouses often requires intensive pesticide use, which raises serious concerns over food safety and human health. This study investigated uptake, translocation and residue dissipation of typical fungicides (metalaxyl-M and fludioxonil) and insecticides (cyantraniliprole and thiamethoxam) in greenhouse chrysanthemum when applied in soils. Chrysanthemum plants could absorb these pesticides from soils via roots to various degrees, and bioconcentration factors (BCFLS) were positively correlated with lipophilicity (log Kₒw) of pesticides. Highly lipophilic fludioxonil (log Kₒw = 4.12) had the greatest BCFLS (2.96 ± 0.41 g g⁻¹), whereas hydrophilic thiamethoxam (log Kₒw = −0.13) had the lowest (0.09 ± 0.03 g g⁻¹). Translocation factors (TF) from roots to shoots followed the order of TFₗₑₐf > TFₛₜₑₘ > TFfₗₒwₑᵣ. Metalaxyl-M and cyantraniliprole with medium lipophilicity (log Kₒw of 1.71 and 2.02, respectively) and hydrophilic thiamethoxam showed relatively strong translocation potentials with TF values in the range of 0.29–0.81, 0.36–2.74 and 0.30–1.03, respectively. Dissipation kinetics in chrysanthemum flowers followed the first-order with a half-life of 21.7, 5.5, 10.0 or 8.2 days for metalaxyl-M, fludioxonil, cyantraniliprole and thiamethoxam, respectively. Final residues of these four pesticides, including clothianidin (a primary toxic metabolite of thiamethoxam), in all chrysanthemum flower samples were below the maximum residue limit (MRL) values 21 days after two soil applications each at the recommended dose (i.e., 3.2, 2.1, 4.3 and 4.3 kg ha⁻¹, respectively). However, when doubling the recommended dose, the metabolite clothianidin remained at concentrations greater than the MRL, despite that thiamethoxam concentration was lower than the MRL value. This study provided valuable insights on the uptake and residues of metalaxyl-M, fludioxonil, cyantraniliprole and thiamethoxam (including its metabolite clothianidin) in greenhouse chrysanthemum production, and could help better assess food safety risks of chrysanthemum contamination by parent pesticides and their metabolites.

During their lifetime honey bees (Apis mellifera) rarely experience optimal conditions. Sometimes, a simultaneous action of multiple stressors, natural and chemical, results in even greater effect than of any stressor alone. Therefore, integrative investigations of different factors affecting honey bees have to be carried out. In this study, adult honey bees exposed to thiamethoxam in larval and/or adult stage and infected with Nosema ceranae were examined. Newly emerged bees from colonies, non-treated or treated with thiamethoxam, were organized in six groups and kept in cages. Thiamethoxam treated bees were further exposed to either thiamethoxam or Nosema (groups TT and TN), or simultaneously to both (group TTN). Newly emerged bees from non-treated colonies were exposed to Nosema (group CN). From both, treated and non-treated colonies two groups were organized and further fed only with sugar solution (groups C and TC). Here, we present the expression profile of 19 genes in adult worker honey bees comprising those involved in immune, detoxification, development and apoptosis response. Results showed that gene expression patterns changed with time and depended on the treatment. In group TC at the time of emergence the majority of tested genes were downregulated, among which nine were significantly altered. The same gene pattern was observed on day six, where the only significantly upregulated gene was defensin-1. On day nine most of analyzed genes in all experimental groups showed upregulation compared to control group, where upregulation of antimicrobial peptide genes abaecin, defensin-1 and defensin-2 was significant in groups TT and TTN. On day 15 we observed a similar pattern of expression in groups TC and TT exposed to thiamethoxam only, where most of the detoxification genes were downregulated. Additionally RNA loads of Nosema and honey bee viruses were recorded. We detected a synergistic interaction of thiamethoxam and Nosema, reflected in lowest honey bee survival.

Neonicotinoids and the closely related insecticide classes sulfoximines and butenolides have recently attracted growing concerns regarding their potential negative effects on non-target organisms, including pollinators such as bees. Indeed, it is becoming increasingly clear that these effects may occur at much lower levels than those considered to be safe for humans. To properly assess the ecological and environmental risks posed by neonicotinoids, appropriate sampling and analytical procedures are needed. Here, we used honey as reliable environmental sampler and developed an unprecedentedly sensitive method based on QuEChERS and UHPLC-MS/MS for the simultaneous determination of the nine neonicotinoids and related molecules currently present on the market (acetamiprid, clothianidin, dinotefuran, flupyradifurone, imidacloprid, nitenpyram, sulfoxaflor, thiacloprid and thiamethoxam). The method was validated and provided excellent levels of precision and accuracy over a wide concentration range of 3–4 orders of magnitude. Lowest limits of quantification (LLOQs) as low as 2–20 pg/g of honey depending on the analytes were reached. The method was then applied to the analysis of 36 honey samples from various regions of the World which had already been analysed for the five most common neonicotinoids (acetamiprid, clothianidin, imidacloprid, thiacloprid and thiamethoxam) in a previous study. This allowed us to determine the long-term stability (i.e. up to 40 months) of these molecules in honey, both at room temperature and −20 °C. We found that the five pesticides were stable over a period of several years at −20 °C, but that acetamiprid and thiacloprid partially degraded at room temperature. Finally, we also measured the levels of dinotefuran, nitenpyram, sulfoxaflor and flupyradifurone and found that 28% of the samples were contaminated by at least one of these pesticides.