Neurotoxicity of tetramethylammonium ion on larval and juvenile zebrafish: Effects on neurobehaviors and multiple biomarkers.

Tetramethylammonium hydroxide (TMAH) is an important compound that utilized and released by the rapidly expanding semiconductor industry, which could hardly be removed by the conventional wastewater treatment techniques. As a cholinergic agonist, the tetramethylammonium ion (TMA+) has been reported to induce toxicity to muscular and respiratory systems of mammals and human, however the toxicity on aquatic biota remains poorly known. We investigated the neurotoxic effects of TMA+ exposure on zebrafish, based on neurobehavior tests and a series of biomarkers. Significant inhibitions on the swimming distance of zebrafish larvae were observed when the exposure level exceeded 50 mg/L, and significant alterations on swimming path angles (straight and deflective movements) occurred even at 10 mg/L. The tested neurobehavioral endpoints of zebrafish larvae were significantly positively correlated with reactive oxygen species (ROS) and malondialdehyde (MDA), significantly negatively related with the activities of antioxidant enzymes, but not significantly correlated with the level of acetylcholinesterase (AChE). Such relationship indicates that the observed neurotoxic effects on swimming behavior of zebrafish larvae is mainly driven by oxidative stress, rather than the alterations of neurotransmitter. At the highest exposure concentration (200 mg/L), TMA+ evoked more severe toxicity on zebrafish juveniles, showing significantly stronger elevation on the MDA activity, and greater inhibitions on the activities of antioxidant enzymes and AChE, suggesting juveniles were more susceptible to TMA+ exposure than larval zebrafish.

Variations in Hydraulic Conductivity of Montmorillonite in Dual-Cation Electrolyte Solutions.

The hydraulic conductivity of Na-montmorillonite in dual-cation solutions of Na+ and Mn+ (Mn+ = K+, Ca2+, Zn2+ and Al3+) with a constant ionic strength of 0.1 mol/L was determined. The focus of this study was on the influence of Mn+ on the grain-size distribution of montmorillonite and hence its hydraulic conductivity. All the tested cations showed a high affinity towards montmorillonite, and the high valency favored the exchange between Mn+ and Na+. The hydraulic conductivity of montmorillonite increased to the maxima and then decreased in a left-skewed log-normal shape as the cation exchange progressed. The grain size of montmorillonite concurrently decreased monotonically with the cation exchange. The XRD patterns of montmorillonite confirmed the occurrence of demixing of Na+ and K+ in the interlayers. It is proposed that the rearrangement and reaggregation of grains during cation exchange occurred, leading to variations in the hydraulic conductivity of montmorillonite.

Lindane degradation in wet-dry cycling soil as affected by aging and microbial toxicity of biochar.

This study determined the degradation of lindane in soil amended with biochar to evaluate the effects of biochar aging and microbial toxicity. Two biochars were prepared at 400 and 600 °C (BC400 and BC600) and subjected to acid washing to remove nutrition (WBC400 and WBC600). After 89 days of incubation under the alternate "wet-dry" conditions, scanning electron microscopy showed that acid washing rendered biochars especially susceptible to aging with structural collapse and fragmentation, with less surface covering. Aging impeded the release of toxic substances in BC400 and BC600 with reduced toxicity to degrading microorganisms. Lindane degradation was somewhat stimulated by biochar nutrition but mainly inhibited by adsorption. Acid washing facilitated the release of toxic substances and additionally reduced lindane degradation. The variations in fatty acid saturation degree (SFA/UFA) in soils confirmed the microbial toxicity of 5% WBC400 > 5% BC400 > 5% BC600 > 5% WBC600. High-throughput DNA sequencing showed that biochar delayed the formation of dominant degrading microbial communities in soil. Lindane degradation was completed by joint Sphingomonas, Flaviolibacter, Parasegetibacter, Azoarcus, Bacillus and Anaerolinaea. These findings are helpful for better understanding the effect of biochar in soil on long-term degradation of persistent organic pollutants.

Enhanced removal of cadmium from wastewater with coupled biochar and Bacillus subtilis.

Shortcomings of individual biochar or microbial technologies often exist in heavy metal removal from wastewater and may be circumvented by coupled use of biochar and microorganisms. In this study, Bacillus subtilis and each of three biochars of different origins (corn stalk, peanut shell, and pine wood) were coupled forming composite systems to treat a cadmium (Cd, 50 mg/L) wastewater formulated with CdCl2 in batch tests. Biochar in composite system enhanced the activity and Cd adsorption of B. subtilis. Compared with single systems with Cd removal up to 33%, the composite system with corn stalk biochar showed up to 62% Cd removal, which was greater than the sum of respective single B. subtilis and biochar systems. Further analysis showed that the removal of Cd by the corn stalk composite system could be considered to consist of three successive stages, that is, the biochar-dominant adsorption stage, the B. subtilis-dominant adsorption stage, and the final biofilm formation stage. The final stage may have provided the composite system with the ability to achieve prolonged steady removal of Cd. The biochar-microorganism composite system shows a promising application for heavy metal wastewater treatment.

Relation of tributyltin and triphenyltin equilibrium sorption and kinetic accumulation in carp and Ceratophyllum demersum.

Comparatively limited knowledge is known about the accumulation processes of tributyltin (TBT) and triphenyltin (TPT) in fish and aquatic plant in the freshwater environment, which has hindered a full understanding of their bioaccumulation potential and ecological risks. In the present study, sorption of TBT and TPT on dead biota of both carp and C. demersum from water via the batch equilibrium technique as well as uptake of them on live biota of both carp and C. demersum from water at a static and a dynamic kinetics tests were investigated, respectively. Both TBT and TPT exhibit a high affinity in carps and C. demersum. And C. demersum has a faster metabolism either for TBT or TPT than carp. The apparent uptake values (Cbio = 1904-8831 µg/kg) or bioconcentration factor (BCF = 3333-44000 L/kg) were one or two orders of magnitude higher than that of estimated by a simple sorption (405-472 µg/kg) or lipid model (74.5-149.6 µg/kg) for carp, indicating the uptake of TBT and TPT did not only depend on lipids but also oxygen ligands or macromolecules such as amino acids and proteins of the living organism. In contrast, the apparent Cbio values (149.1-926.4 µg/kg) of both TBT and TPT were lower than that of estimated by sorption model (1341-1902 µg/kg) for C. demersum, which were due to the rapid metabolic rate of them, especially for TBT. But no relation was observed between TBT and TPT concentrations and lipid contents in C. demersum.

Removal of dichlorophenol by Chlorella pyrenoidosa through self-regulating mechanism in air-tight test environment.

Microalgae are surprisingly efficient to remove pollutants in a hermetically closed environment, though its growth is inhibited in the absence of pollutants. The final pH, algal density, Chl-a content, and the removal efficiency of 2,4-dichlorophenol (2,4-DCP) by Chlorellar pyrenoidosa in a closed system were observed under different initial pH, lighting regimes, and various carbon sources. The optimal condition for 2,4-DCP removal was obtained, and adopted to observe the evolution of above items by domesticated and origin strains. The results showed that both respiration and photosynthesis participated in the degradation of 2,4-DCP, and caused the changes of pH. The photosynthesis seemed to increase the solution pH, while the respiration and the biodegradation of 2,4-DCP to decrease the solution pH. The domesticated strain achieved nearly 100% removal when initial concentrations of 2,4-DCP lower than 200 µg L-1, due to providing a appropriate but narrow pH evolution range, mostly falling between 6.5 and 7.9. The research helps to understand the mechanism of biodegradation of chlorophenol compounds by green algae.

Cosolvent Effects on Dechlorination of Soil-Sorbed Polychlorinated Biphenyls Using Bentonite Clay-Templated Nanoscale Zero Valent Iron.

Zero-valent iron synthesized using bentonite clay as a template (CZVI) was tested for its reactivity toward polychlorinated biphenyl (PCB) dechlorination in soil slurries. Aqueous-phase decachlorobiphenyl (PCB209) was rapidly dechlorinated by CZVI with a reaction rate 10 times greater than that by conventional nanoscale zerovalent iron. This superior reactivity was due largely to the nanoscale size (∼0.5 nm) of the ZVI particles located in the clay galleries. In soil slurries where PCB209 was strongly soil-bound, adding ethanol as an organic cosolvent led to increased PCB209 desorption into the liquid phase, thereby enhancing the PCB209 dechlorination with CZVI. The more effective PCB209 dechlorination in such a cosolvent system also promoted the subsequent stepwise dechlorinative process, leading to a relatively more removal of chlorine in the product mixture. The dechlorination became more rapid as the ethanol fraction increased from 10% to 50%, due apparently to the increasingly greater PCB209 desorption and thus facilitated contact with CZVI. Further increase in ethanol fraction above 50% led to an insignificant enhancement in degradation rate, due partially to attenuated contact of PCB209 with CZVI and reduced proton source from limited water content in the liquid. It is suggested that addition of organic cosolvents may make CZVI potentially useful for remediation of soils containing halogenated organic contaminants.

Internalization and Phytotoxic Effects of CuO Nanoparticles in Arabidopsis thaliana as Revealed by Fatty Acid Profiles.

Internalization and phytotoxic effects of CuO nanoparticles (nCuO) in plants were studied at the cellular level. Arabidopsis thaliana was hydroponically challenged by nCuO (100 mg/L), as compared to Cu2+ ions (1.2 mg/L), to account for nCuO dissolution for 96 h and 28 days to monitor Cu accumulation in the plant as well as the fatty acid (FA) profiles of the plant cell membrane. Under the same growing conditions, the nCuO exposure resulted in more Cu accumulation than did the Cu2+ exposure. Multiple microscopic techniques confirmed the internalization and sequestration of nCuO in root cell vacuoles, where transformation of Cu(II) to Cu(I)Cl occurred. Short and long exposures (96 h versus 28 days) to both nCuO and Cu2+ elevated FA saturation degrees in plant cells through oxidative stress, as verified by in situ detection of superoxide radicals, with conversions mostly from C18:3, C16:3, and C18:2 to C16:0. Only the long exposure to nCuO significantly brought about an additional elevation of FA saturation degree in root cells. These results demonstrated that the acute effects of plant exposure to nCuO were mainly produced from the stress of Cu2+ ions released from nCuO dissolution, while the chronic effects in roots were significantly developed by the nCuO particle stress. The findings in this work are novel and may offer significant implications in better understanding nanoparticle-induced phytotoxicity and potential risks in ecosystems.

Distribution and spatial variation of volatile methylsiloxanes in surface water and wastewater from the Yangtze River Basin, China.

Volatile methylsiloxanes (VMSs) earned serious concerns due to their detection and toxicities after their release to the environments. They were also detected in rivers around the globe, but their distribution remained to be explored in larger rivers with longer length, higher water volume and wider watershed. In the present study, 8 cyclic VMSs (cVMSs) and 7 linear ones (lVMSs) were investigated in 42 water samples (27 surface water (including 7 drinking source water) and 15 wastewater) from the Yangtze River Basin, China. Results showed that VMSs were detected in all sampling sites. In surface water, the concentrations of total cVMSs ranged from 17.3 to 4.57 × 103 ng/L, while those of lVMSs ranged from 1.72 to 81.6 ng/L. In wastewater, the total concentrations of cVMSs and lVMSs showed ranges of 17.6-1.66 × 103 ng/L and 2.59-252 ng/L, respectively. Apparently, cVMSs showed significantly higher concentrations than lVMSs. The concentrations of cVMSs followed an order of lower > upper > middle reaches, while those of lVMSs did not show clear distribution patterns. Among cVMSs, those with less Si numbers were dominant, while those with more Si numbers were dominant in lVMSs. Notably, the VMSs were also detected in 7 surface waters that served as drinking source waters, which earned them further concerns. In addition, the VMSs in surface water showed positive correlation with those in wastewater, which led to necessity in management on industrial emissions in the future.

Geochemical controls on the distribution and bioavailability of heavy metals in sediments from Yangtze River to the East China Sea: Assessed by sequential extraction versus diffusive gradients in thin-films (DGT) technique.

This study conducted a comprehensive investigation on the distribution and bioavailability of heavy metals (Cr, Co, Ni, Cu, Zn, Cd and Pb) in sediments along two typical transects from Yangtze River to the East China Sea continental shelf that spanning large physicochemical gradients. Heavy metals were mainly associated with the fine-grained sediments (enriched with organic matter), exhibiting decreasing trends from nearshore to offshore sites. The turbidity maximum zone showed the highest metal concentrations, which evaluated as polluted for some tested metals (especially Cd) using the geo-accumulation index. Based on the modified BCR procedure, the non-residual fractions of Cu, Zn and Pb were higher within the turbidity maximum zone, and significantly negatively correlated with bottom water salinity. The DGT-labile metals all positively correlated with the acid-soluble metal fraction (especially for Cd, Zn and Cr), and negatively correlated with salinity (except Co). Therefore, our results suggest salinity as the key factor controlling metal bioavailability, which could further modulate metal diffusive fluxes at the sediment-water interface. Considering that DGT probes could readily capture the bioavailable metal fractions, and reflect the impacts of salinity, we suggest DGT technique can be used as a robust predictor for metal bioavailability and mobility in estuary sediments.

Altered transfer of heavy metals from soil to Chinese cabbage with film mulching.

The influence of film mulching on the migration of metals from soil to cabbage was investigated. Following a 50-day growth in field plots mulched or unmulched, root-zone soils and Chinese cabbage (Brassica chinensis L.) were sampled for metal analysis. Mulching slightly decreased the soil mobile (acid-extractable) Cd, but increased its transfer from root to the cabbage parts. As an essential element, Cu was readily transferred to the cabbage parts. While mulching decreased the soil mobile Zn, reduced soil pH resulted in its enhanced soil-to-root migration. This, however, did not increase the transfer of Zn within cabbage. Although mulching increased the soil mobile Pb by 200%, an increase in Pb in cabbage leaves but a decrease in stem result presumably from the enhanced foliar uptake of atmospheric Pb. This study suggests that mulching may promote the accumulation of toxic metals such as Cd and Pb in cabbage and therefore increase crop risks to human health.

Lindane uptake and translocation by rice seedlings (Oryza sativa L.) under different culture patterns and triggered biomass re-allocation.

The study was conducted to investigate the influence of the culture pattern on plant uptake and translocation of an organic chemical and the resultant acute response of plants, and to further reveal the interconnection. Plant exposure experiments were performed using a conventional rice seedling (Oryza sativa L. subsp. indica) under two kinds of culture patterns (viz., hydroponics and soil-based culture) with various culture matrices for a period of 7 days. The exposure concentration of lindane was ∼450 µg L-1 in the aqueous-phase matrices, and 200.1-756.0 µg kg-1 in the solid matrices. Lindane accumulation and its distribution in plant tissues were quantified, as well as the tissue biomass. The results showed the accumulation of lindane in all exposure groups were comparatively close over the period, confirming that the soil-bound lindane was scarcely available to plants. Similar trend of lindane uptake and translocation in seedlings was found among the groups under the same kind of cultivation pattern. In the hydroponic groups, lindane was mostly distributed in roots (about 60% at the end of exposure), whereas more lindane was translocated to shoots (approximate 70%) under the soil-based culture pattern. Allometric analysis demonstrated that the tissue part (root or shoot) with more lindane accumulation had a relatively higher growth rate over 7 days. Correspondingly, biomass allocation presented a slight trend of mutual proximity to lindane distribution. It was inferred that plants altered their allometric growth pattern to realize biomass re-allocation in response to the short-term lindane exposure, which could be considered as a plant defense strategy.

Blocking effect of fullerene nanoparticles (nC60) on the plant cell structure and its phytotoxicity.

Blockage of nanoparticles on plant pore structures might produce phytotoxicity and affect plant uptake indirectly. This study examined the blocking and phytotoxic effects of fullerene nanoparticles (nC60) on plants at the cellular level. The malondialdehyde content in plant was normal during nC60 exposure, implying that nC60 caused no acute phytotoxicity, while the normalized relative transpiration significantly decreased, showing that the pore structure of roots was seriously blocked by nC60. High power optical microscopy and transmission electron microscope showed that root endothelial cells were squeezed, and inner wall structures were damaged by the extrusion of nanoparticles. Low nC60 concentrations inhibited root uptake of lindane, whereas high nC60 concentrations promoted root uptake of lindane, indicating that serious pore blocking by nC60 damaged root cell structure and hence ready transport of lindane from roots to shoots. Significant alterations of fatty acid (FA) saturation degree of root cell membrane indicated that nC60 led to phytotoxicity in the root cell membrane after long-term exposure and nC60 produced phytotoxicity in the process of blocking root pore structures and interfering with cell membrane fluidity. Moreover, the plant cell structures under phytotoxicity were more likely to be damaged mechanically by the extrusion of nanoparticles. These findings may be helpful to better understand the transport pathways of nanoparticles in plants, the phytotoxicity of nanoparticles and the potential risks of nanomaterials used in agriculture.

Translocation versus ion trapping in the root uptake of 2,4-dichlorophenol by wheat seedlings.

Understanding of the plant uptake of ionizable organic compounds is critical to the evaluation of crop contamination, plant protection, and phytoremediation. This study investigated the time-dependent uptake of 2,4-dichlorophenol (DCP) by intact wheat seedling roots and subsequent translocation to shoots at pH 5.0 and 8.0. Sorption of DCP by cut roots and shoots at these two pHs was conducted to provide the uptake limits and the Donnan charge. For comparison, sorption was also conducted for 1,3-dichlorobenzene (DCB), a nonionizable compound having a structure similar to that of DCP. The DCB sorption isotherms were linear and independent of pH, yielding a consistent log Klip of 3.56 with both roots and shoots, reflective of the essential dominant role of lipids in plant partition uptake. Whereas the DCP sorption also showed a linear isotherm at pH 5.0 with log Klip = 2.88, the sorption at pH 8.0 was nonlinear with a concave downward shape, especially at low concentrations. With live wheat seedlings, the DCB uptake by roots and the DCB translocation to shoots rapidly approached a steady state, showing no obvious pH effect. On the DCP uptake by live plants, there was a rapid attainment of a steady state in roots at pH 5.0 coupled with a retarded transport to shoots due presumably to the polarity of DCP. At pH 8.0, the root uptake of DCP was comparatively slower and the translocation to shoots was completely inhibited due presumably to DCP ionization. At high pH, DCP was supposedly accumulated in an ionized form in root cells via an ion-trapping mechanism.

Mechanisms of aromatic molecule - Oxygen-containing functional group interactions on carbonaceous material surfaces.

Surface oxygen-containing functional groups (OFGs) at different sites of carbonaceous materials showed different effects on the normalized monolayer adsorption capacity (QBET/A) obtained from the modified BET model. The OFGs on mesoporous surfaces inhibited the adsorption via the water competition, whereas those on the external surface promoted the adsorption due to the enhanced hydrophobic driving force and electrostatic forces, as analyzed from the adsorption molar free energy. Multiple linear relationships were established between the monolayer adsorption capacity QBET/A and the amounts of OFGs on mesoporous and the external surfaces ([O]meso and [O]external, respectively). The properties of aromatic adsorbate compounds, the polar area radio of aromatic molecule to water (PAad/w), and the log Kow together influenced the inhibition or promotion effects of OFGs. These results would allow predictions of adsorption behavior of aromatic compounds on carbonaceous materials on the basis of OFGs parameters. Theoretical calculations and simulations projected the configuration of aromatic molecules being parallel to the graphene sheets of carbonaceous materials. The symmetry-adapted perturbation theory (SAPT) energy decomposition showed that the electrostatic forces intensified with the increase of adsorbate polarity. These analyses revealed that the electrostatic forces were enhanced in the presence of OFGs and that the π-π EDA (electron donor-acceptor) was the main force.

Microplastics combined with tetracycline in soils facilitate the formation of antibiotic resistance in the Enchytraeus crypticus microbiome.

Growing evidence suggests that microplastics can adsorb antibiotics and may consequently exacerbate effects on the health of exposed organisms. Our current understanding of the combined effects of microplastics and antibiotics on antibiotic resistance genes (ARGs) in soil invertebrates is limited. This study aimed to investigate changes in the microbiome and ARGs in Enchytraeus crypticus following exposure to a soil environment that contained both microplastics and antibiotics. Tetracycline (TC), polyamide (PA) and polyvinyl chloride (PVC) were used to construct microcosms of polluted soil environments (TC, PA, PVC, PA+TC, PVC+TC). The differences in microbiomes and ARGs were determined by bacterial 16S rRNA gene amplicon sequencing and high throughput quantitative PCR. The results show that compared with the Control or microplastics alone treatments, TC was significantly accumulated in E. crypticus when exposed to TC alone or in combination with microplastics (P < 0.05), but there were no significant differences about TC accumulation between TC, PA+TC, and PVC+TC treated E. crypticus (P > 0.05). Microplastics and TC significantly disturbed the microbial community, and decreased the microbial alpha diversity of E. crypticus (P < 0.05). However, there were no significant differences between TC, microplastics and their combined exposure treatments, and no toxic synergies on the diversity of E. crypticus microbiome between tetracycline and microplastics in soil environment. All the treatments increased the diversity of ARGs in E. crypticus (39-49 ARGs vs. 25 ARGs of control). In particular, treatments combining PVC and TC or PA and TC exposure resulted in greater ARGs abundance than the treatments when E. crypticus was exposed to PVC, PA or TC alone. These results add to our understanding of the combined effects of microplastics and antibiotics on the ARGs and microbiome of soil invertebrates.

Uptake and elimination of butyl- and phenyltins by Ceratophyllum demersum L.

The widespread occurrence and distribution of organotin compounds (OTCs) in both marine and freshwater ecosystems has aroused considerable concerns in most countries worldwide. In this work, individual kinetics of the elimination of three butyltins and three phenyltins from C. demersum L. were systematically studied for over 240 h in clean water after a 48h period of accumulation. All OTCs were rapidly metabolized to nontoxic inorganic tin by C. demersum L. through stepwise debutylation or dephenylation. In addition to inorganic tin, monobutyltin (MBT) and monophenyltin (MPT) were the primary degradation products of tributyltin (TBT) and triphenyltin (TPT), with small amounts of dibutyltin (DBT) and diphenyltin (DPT), respectively, also being present. The estimated half-life of TPT (240 h) in C. demersum L. was longer than that of TBT (100 h), although the TPT was less hydrophobic. The corresponding degradation mechanisms may be attributed to a cascade of enzymatic reactions of CYP450 enzymes in C. demersum L. The pH played an important role in both plant growth and TBT degradation. Although pH 8.9 was more suitable for C. demersum L. growth, it uptook and metabolized more TBT at pH 5.0, which may be because the cationic species TBT+ (at pH 5.0) was metabolized more easily than the neutral hydroxide species TBTOH (at pH 8.9). C. demersum L. may thus be the plant with the most potential for the remediation of OTC-contaminated freshwater environments.

The long-term environmental risks from the aging of organochlorine pesticide lindane.

Although increased contact time (aging) of pesticides in the soil decreases their bioavailability, this does not mean that the bound residues formed during the aging process pose fewer risk to the soil environment. Here the earthworm Eisenia fetida was exposed to organochlorine pesticide lindane in soil under different durations of lindane aging and exposure. The results of de novo RNA sequencing followed by molecular and biochemical validations demonstrated the aged lindane showed a different tendency to disrupt acetylcholine (ACh) transmission with the effects of fresh lindane to gamma-aminobutyric acid. Using own-developed earthworm activity test, we confirmed aged lindane prompted earthworms to exclusively exhibit a significant hypoactivity in locomotion, which could be explained by the inhibition of Ach system. This study suggested that the toxicity of pesticides would not depend solely on their free state components, and the awareness of long-term environmental risks from the bound states needs to be raised.

Do combined nanoscale polystyrene and tetracycline impact on the incidence of resistance genes and microbial community disturbance in Enchytraeus crypticus?

It has been proved that nanoplastics can effectively adsorb pollutants and thus influence their behavior and availability. The combined toxic effects of nanoplastic and its adsorbed pollutant on the soil fauna are still not well known. We used high-throughput quantitative PCR to explore the effects of oral nanoscale polystyrene and tetracycline exposure on antibiotic resistance genes in the soil invertebrate Enchytraeus crypticus, and used bacterial 16S rRNA gene amplification sequencing to examine the response of the microbiome of E. crypticus. After 14 days of tetracycline and nanoscale polystyrene exposure, we terminated exposure and monitored the restoration of ARGs and microbiome in the E. crypticus. Results showed that the number of ARGs, especially macrolide-lincosamide-streptogramin B (MLSB), tetracycline ARGs, as well as multidrug ARGs, increased with exposure to nanoscale polystyrene and tetracycline. The abundance of Aminoglycoside and Beta_Lactamase ARGs in E. crypticus also significantly increased. The exposure significantly perturbed the abundance of families Microbacteriaceae, Streptococcaceae, Enterobacteriaceae, Rhodocyclaceae and Sphinomonadaceae. After terminating exposure for 14 days, the diversity and abundance of ARGs were not completely restored, while the microbiome was not permanently changed but reversibly impacted.

Does soil CuO nanoparticles pollution alter the gut microbiota and resistome of Enchytraeus crypticus?

Growing evidence suggests that metallic oxide nanoparticles can pose a severe risk to the health of invertebrates. Previous attention has been mostly paid to the effects of metallic oxide nanoparticles on the survival, growth and physiology of animals. In comparison, the effects on gut microbiota and incidence of antibiotic resistance genes (ARGs) in soil fauna remain poorly understood. We conducted a microcosm study to explore the responses of the non-target soil invertebrate Enchytraeus crypticus gut microbiota and resistomes to copper oxide nanoparticles (CuO NPs) and copper nitrate by using bacterial 16S rRNA gene amplicons sequencing and high throughput quantitative PCR. The results showed that exposure to Cu2+ resulted in higher bioaccumulation (P < 0.05) and lower body weight and reproduction (P < 0.05) of Enchytraeus crypticus than exposure to CuO NPs. Nevertheless, exposure to CuO NPs for 21 days markedly increased the alpha-diversity of the gut microbiota of Enchytraeus crypticus (P < 0.05) and shifted the gut microbial communities, with a significant decline in the relative abundance of the phylum Planctomycetes (from 37.26% to 19.80%, P < 0.05) and a significant elevation in the relative abundance of the phyla Bacteroidetes, Firmicutes and Acidobacteria (P < 0.05). The number of detected ARGs in the Enchytraeus crypticus gut significantly decreased from 45 in the Control treatment to 16 in the Cu(NO3)2 treatment and 20 in the CuO NPs treatment. The abundance of ARGs in the Enchytraeus crypticus gut were also significantly decreased to 38.48% when exposure to Cu(NO3)2 and 44.90% when exposure to CuO NPs (P < 0.05) compared with the controls. These results extend our understanding of the effects of metallic oxide nanoparticles on the gut microbiota and resistome of soil invertebrates.