International audience | We assessed the state of knowledge regarding the effects of large-scale pollution with neonicotinoid insecticides and fipronil on non-target invertebrate species of terrestrial, freshwater and marine environments. A large section of the assessment is dedicated to the state of knowledge on sublethal effects on honeybees (<em>Apis mellifera</em>) because this important pollinator is the most studied non-target invertebrate species. Lepidoptera (butterflies and moths), Lumbricidae (earthworms), Apoidae sensu lato (bumblebees, solitary bees) and the section “other invertebrates” review available studies on the other terrestrial species. The sections on freshwater and marine species are rather short as little is known so far about the impact of neonicotinoid insecticides and fipronil on the diverse invertebrate fauna of these widely exposed habitats. For terrestrial and aquatic invertebrate species, the known effects of neonicotinoid pesticides and fipronil are described ranging from organismal toxicology and behavioural effects to population-level effects. For earthworms, freshwater and marine species, the relation of findings to regulatory risk assessment is described. Neonicotinoid insecticides exhibit very high toxicity to a wide range of invertebrates, particularly insects, and field-realistic exposure is likely to result in both lethal and a broad range of important sublethal impacts. There is a major knowledge gap regarding impacts on the grand majority of invertebrates, many of which perform essential roles enabling healthy ecosystem functioning. The data on the few non-target species on which field tests have been performed are limited by major flaws in the outdated test protocols. Despite large knowledge gaps and uncertainties, enough knowledge exists to conclude that existing levels of pollution with neonicotinoids and fipronil resulting from presently authorized uses frequently exceed the lowest observed adverse effect concentrations and are thus likely to have large-scale and wide ranging negative biological and ecological impacts on a wide range of non-target invertebrates in terrestrial, aquatic, marine and benthic habitats.

The social and ecological influence of Neonicotinoids (NEOs) usage in agriculture sector is progressively higher. There are seven NEOs insecticides widely used for the insects control. Among the NEOs, thiacloprid (THD) was extensively used for insect control during crop cultivation. This study targets to analyse the contamination levels of NEOs in agricultural soil and identify photo-biodegradation of THD degradation using pure isolates and mixed consortium. The photo degradation (PD), biodegradation (BD) and photo-biodegradation (PBD) of THD were compared. The corn field agricultural soils were polluted by four NEOs, among them THD had greater contamination level (surface soil: 3901.2 ± 0.04 μg/g) and (sub-surface soil: 3988.6 ± 0.05 μg/g). Three soil free enriched bacterial strains following Bacillus atrophaeus (PB-2), Priestia megaterium (PB-3) (formerly known as Bacillus megaterium), and Peribacillus simplex (PB-4) (formerly known as Bacillus simplex) were identified by microbiological and molecular 16s rRNA gene sequencing. The PD, BD and PBD of THD were conducted and degradation rate was detected by instrument UPLC-MS-MS. The PBD process with blue-LEDs showed better THD degradation efficiency than PD and BD, where the specific THD degradation rate was 85 ± 0.2%, 87 ± 0.5%, and 89 ± 0.3%, respectively for PB-2, PB-3 and PB-4. Then, the photo-biodegradation performance is greater at 150, 175, 200 rpm, pH 7.0–9.0, and temperature 30–35 °C. After the PBD system deliver four intermediate metabolites, the THD degradation process maybe through nitro reduction, hydroxylation and oxidative cleavage pathway.

We investigated the fates of seven neonicotinoids (NNIs) in full-scale drinking water treatment plants and assessed human exposure to NNIs through consuming drinking water. The total NNI concentrations in raw water and treated water samples from the drinking water treatment plants were 20.4–166 ng/L (median 118 ng/L) and 1.11–94.7 ng/L (median 20.4 ng/L), respectively. The dinotefuran (DIN) concentrations in raw water collected in different seasons were different, and the highest DIN concentration was found in summer. The drinking water treatment processes removed >91% of the NNIs except DIN and thiamethoxam (THIAM), for which the mean removal rates were 70% and 74%, respectively. The removal rates for all of the NNIs were higher for the granular activated carbon filtration process (mean 83.5%) than the other drinking water treatment plant processes (coagulation/sedimentation 22.3%, ozonation 29.2%). However, the removal rates in the granular activated carbon process were lower for DIN and THIAM (61.0% and 59.2%, respectively) than the other NNIs. Significant correlations were found between the NNI removal rates and physicochemical properties (solubility in water and log (octanol–water partition coefficient)). The estimated mean human exposure to NNIs in drinking water was 0.528 ng/(kg body weight d).

The extensive use of neonicotinoids (NEOs) has caused the release of wide-ranging of residues to the environment and food, and their potential health risks are now receiving more attention. In this study, three surveys were conducted to obtain the overall profiles of NEO residue levels (seven NEOs and one metabolite) in Chinese tea over a period of seven years. A total of 726 tea samples were tested, and nearly 87% of the samples were found to have detectable NEO residues. The overall average detection frequency of acetamiprid was the highest, reaching 73%. Imidacloprid residues in 4.6% of the samples exceeded the Chinese maximum residue limits, whereas clothianidin and nitenpyram had been detected in Chinese tea samples since 2014. The applications of thiacloprid and thiamethoxam gradually increased, and some tea samples with high residue levels appeared in China. These findings signal the replacement of new and old varieties of NEOs in China. Both long- and short-term cumulative exposures to NEOs were calculated based on optimistic and pessimistic models recommended in the EFSA guidelines. In the three survey periods, the average total imidacloprid-equivalent concentrations were 484.63, 1713.36, and 1148.34 μg/kg, respectively. Combined with the refined point estimates and probabilistic models used in this study, the hazard quotients of NEO residues in tea for Chinese tea consumers were found to be low and within the bounds of safety.

The ecological impacts of insecticides in aquatic areas around agricultural lands have long been ignored in the regulation scheme of pesticides in Japan. Upon the scheme, the predicted concentration of an insecticide in the main stream of a river is the only parameter considered, suggesting that the ecological impacts of insecticides on local biodiversity around agricultural fields are underestimated. To fill this knowledge gap, we measured insecticide concentrations in surface water and sediment in aquatic areas around paddy fields at 35 locations across Japan. Among the 18 insecticides considered, 15 were detected somewhere in Japan and their concentrations were generally higher in the southwestern region in Japan (e.g. Hiroshima, Saga, or Kagoshima prefectures). Most insecticides were accumulated at higher concentrations in sediment than in surface water, consistent with previous studies. We also detected insecticides applied to nursery boxes at high concentrations in surrounding aquatic areas, although such application is generally considered to have low environmental risks. In addition, derivatives of fipronil, which have similar toxicity as that of fipronil, were often detected in sediment at higher concentrations than fipronil itself. Concentrations of dinotefuran in water at two sampling points were higher than the 5% hazardous concentration (HC5), indicating a possibility of a risk of acute toxicity to aquatic organisms. Our findings indicate that ecological risk assessments of insecticides and their derivatives should be expanded to include concentrations in sediment and water around paddy fields as well.

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.

China is the largest beekeeping and honey consumption country globally. Neonicotinoids in honey can pose adverse effects on honey bees and human, but data on neonicotinoids residues in honey and its health risk remain limited in China. A total of 94 honey samples were selected from Chinese market based on production region and sale volume in 2020. Eight neonicotinoids and four metabolites were determined by liquid chromatography coupled to mass spectrometry. Health risk of neonicotinoids in honey on honey bees and human was assessed by hazard quotient (HQ) and hazard index (HI). Neonicotinoids and their metabolites were overall detected in 97.9% of honey samples. Acetamiprid, thiamethoxam, and imidacloprid were top three dominant neonicotinoids in honey with the detection frequencies of 92.6%, 90.4%, and 73.4%, respectively. For honey bees, 78.7% of honey samples had a HI larger than one based on the safety threshold value of sublethal effects. Top three neonicotinoids with the highest percent proportion of HQ larger than one for honey bees were acetamiprid (43.6%), imidacloprid (31.9%), and thiamethoxam (24.5%) and their maximum HQs were 420, 210, and 41, respectively. Based on oral median lethal doses for honey bees, both HQ and HI were lower than one in all honey samples. For human, both HQ and HI were lower than one based on acceptable daily intakes in all honey samples. Neonicotinoids concentrations and detection frequencies in honey samples and its health risk varied with production region, commercial value of nectariferous plants, number of nectariferous plants, and sale price. The results suggested extensive residues of neonicotinoids in honey in Chinese market with a variation by the characteristics of honey. The residues were likely to affect the health of honey bees, but showed no detectable effect on human health.

In developing countries, urban areas may be at greater risk of pesticide exposure compared to semi-urban agricultural regions. To investigate this, concentrations of selected pesticides were measured in 81 human milk samples collected in urban Kolkata and semi-urban Nadia in West Bengal, India. Three classes of pesticides were investigated – legacy organochlorines and emerging pyrethroids and neonicotinoids. The average concentration of the majority of the chemicals (DDT, its metabolites, HCH isomers, bifenthrin, endosulfan), showed a clear urban > semi-urban trend. Compared with previous measurements in other Indian cities and developing nations, current HCH and DDT concentrations in urban Kolkata were high. These chemicals were detected in 100% of the samples in both the urban and the semi-urban region. Also in both regions, the Estimated Daily Intake of DDTs, HCHs, aldrin, dieldrin and the pyrethroid bifenthrin for breastfed infants exceeded the Tolerable Daily Intake in a number of samples. Three pyrethroids were detected in human milk samples in India for the first time. This indicates a shift in the usage pattern of pesticides in India from organochlorines to pyrethroids. These findings may be used to drive targeted regulation of pesticides in developing countries with similar histories of pesticide use.

Methane emitted by insects is considered to be an important source of atmospheric methane. Here we report the stimulation of methane emission in the cockroach Periplaneta americana and termite Coptotermes chaohuensis, insects with abundant methanogens, by neonicotinoids, insecticides widely used to control insect pests. Cycloxaprid (CYC) and imidacloprid (IMI) caused foregut expansion in P. americana, and increased the methane emission. Antibiotics mostly eliminated the effects. In P. americana guts, hydrogen levels increased and pH values decreased, which could be significantly explained by the gut bacterium community change. The proportion of several bacterium genera increased in guts following CYC treatment, and two genera from four could generate hydrogen. Hydrogen is a central intermediate in methanogenesis. All increased methanogens in both foregut and hindgut used hydrogen as electron donor to produce methane. Besides, the up-regulation of mcrA, encoding the enzyme for the final step of methanogenesis suggested the enhanced methane production ability in present methanogens. In the termite, hydrogen levels in gut and methane emission also significantly increased after neonicotinoid treatment, which was similar to the results in P. americana. In summary, neonicotinoids changed bacterium community in P. americana gut to generate more hydrogen, which then stimulated gut methanogens to produce and emit more methane. The finding raised a new concern over neonicotinoid applications, and might be a potential environmental risk associated with atmospheric methane.

Soil contaminants can cause direct harm to lizards due to their regular swallowing of soil particles. As the world’s fastest growing insecticide with long half-life in soil, the endocrine disrupting effect of neonicotinoids on lizards deserves more attention. In this report, we assessed the endocrine disrupting effect of imidacloprid on Eremias argus during 28 days of continuous exposure. Among the imidacloprid and its metabolites, only the metabolite 6-chloropyridic acid had a significant accumulation in the gonads and was positively correlated with its blood concentration. Imidacloprid might cause endocrine disrupting effects on lizards in two ways. First, the desnitro metabolites of imidacloprid could accumulate in the brain, inhibited the secretion of gonadotropin-releasing hormone, and ultimately affected the feedback regulation of hypothalamic-pituitary-gonadal related hormones. Secondly, imidacloprid severely inhibited the gene expression of the corresponding enzymes in the gonadal anti-oxidative stress system, causing histological damage to the gonads and ultimately affecting gonadal function. Specifically, exposure to imidacloprid resulted in abnormal arrangement of spermatogenic epithelial epithelium, hyperplasia of epididymal wall, and oligospermia of male lizard. Meanwhile, gene expressions of cyp17, cyp19, and hsd17β were severely inhibited in the imidacloprid exposure group, consistent with decreased levels of testosterone and estradiol in plasma. Imidacloprid exposure could cause insufficient androgen secretion and less spermatogenesis in male lizards. The risk of imidacloprid exposure to female lizards was not as severe as that of male lizards, but it still inhibited the expression of cyp19 in the ovaries and led to a decrease in the synthesis of estradiol. This study firstly reported the endocrine disruption of imidacloprid to lizards, providing new data for limiting the use of neonicotinoids.