Intraday variability of indicator and pathogenic viruses in 1-h and 24-h composite wastewater samples: Implications for wastewater-based epidemiology.

We monitored the concentration of indicator viruses crAssphage and pepper mild mottle virus (PMMoV) and human pathogen adenovirus (HAdV) in influent from a wastewater treatment plant in Brisbane, Australia in 1-h and 24-h composite samples. Over three days of sampling, the mean concentration of crAssphage gene copies (GC)/mL in 24-h composite samples did not differ significantly (p = 0.72-0.92), while for PMMoV GC/mL (p value range: 0.0002-0.0321) and HAdV GC/mL (p value range: 0.0028-0.0068) significant differences in concentrations were observed on one day of sampling compared to the other two. For all three viruses, the variation observed in 1-h composite samples was greater than the variation observed in 24-h composite samples. For crAssphage, in 54.1% of 1-h composite samples, the concentration was less than that observed in 24-h composite samples; whereas for PMMoV and HAdV the concentration was less in 79.2 and 70.9% of 1-h composite samples, respectively, compared to the relevant 24-h composite samples. Similarly, the concentration of crAssphage in 1-h compared to 24-h composite samples did not differ (p = 0.1082) while the concentrations of PMMoV (p < 0.0001) and HAdV (p < 0.0001) in 1-h composite samples were significantly different from 24-h composite samples. These results suggest that 24-h composite samples offer increased analytical sensitivity and decreased variability compared to 1-h composite samples when monitoring wastewater, especially for pathogenic viruses with low infection rates within a community. Thus, for wastewater-based epidemiology applications, 24-h composite samples are less likely to produce false negative results and erroneous public health information.

Silver Sulfide Nanoparticles Reduce Nitrous Oxide Emissions by Inhibiting Denitrification in the Earthworm Gut.

The accumulation of Ag2S in agricultural soil via application of Ag-containing sludge potentially affects the functioning of soil microorganisms and earthworms (EWs) due to the strong antimicrobial properties of Ag. This study examined the effects of Ag2S nanoparticles (Ag2S-NPs) on the EW-mediated (Eisenia fetida and Pontoscolex corethrurus) soil N cycle. We used 16S rRNA gene-based sequencing and quantitative polymerase chain reaction to examine the bacterial community and nitrification/denitrification-related gene abundance. The presence of either EWs or Ag significantly increased denitrification and N2O emissions. However, the addition of Ag2S to EW-inhabited soil reduced N2O emissions by 14-33%. Furthermore, Ag2S caused a low-dose stimulation but a high-dose inhibition to N2O flux from the EW gut itself. Accordingly, an increase in Ag in the EW gut caused a decrease in the relative abundance of denitrifiers in both the soil and the gut, especially for the dominant genus Bacillus. Ag2S also decreased the copy numbers of nitrification gene (nxrB) and denitrification genes (napA, nirS, and nosZ) in EW gut, leading to the observed decrease in N2O emissions. Collectively, applying Ag2S-containing sludge disturbs the denitrification function of the EW gut microbiota and the cycling of N in soil-based systems.

Decay of SARS-CoV-2 and surrogate murine hepatitis virus RNA in untreated wastewater to inform application in wastewater-based epidemiology.

Wastewater-based epidemiology (WBE) demonstrates potential for COVID-19 community transmission monitoring; however, data on the stability of SARS-CoV-2 RNA in wastewater are needed to interpret WBE results. The decay rates of RNA from SARS-CoV-2 and a potential surrogate, murine hepatitis virus (MHV), were investigated by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) in untreated wastewater, autoclaved wastewater, and dechlorinated tap water stored at 4, 15, 25, and 37 °C. Temperature, followed by matrix type, most greatly influenced SARS-CoV-2 RNA first-order decay rates (k). The average T90 (time required for 1-log10 reduction) of SARS-CoV-2 RNA ranged from 8.04 to 27.8 days in untreated wastewater, 5.71 to 43.2 days in autoclaved wastewater, and 9.40 to 58.6 days in tap water. The average T90 for RNA of MHV at 4 to 37 °C ranged from 7.44 to 56.6 days in untreated wastewater, 5.58-43.1 days in autoclaved wastewater, and 10.9 to 43.9 days in tap water. There was no statistically significant difference between RNA decay of SARS-CoV-2 and MHV; thus, MHV is suggested as a suitable persistence surrogate. Decay rate constants for all temperatures were comparable across all matrices for both viral RNAs, except in untreated wastewater for SARS-CoV-2, which showed less sensitivity to elevated temperatures. Therefore, SARS-CoV-2 RNA is likely to persist long enough in untreated wastewater to permit reliable detection for WBE application.

Silver Nanoparticles Entering Soils via the Wastewater-Sludge-Soil Pathway Pose Low Risk to Plants but Elevated Cl Concentrations Increase Ag Bioavailability.

The widespread use of silver nanoparticles (Ag-NPs) results in their movement into wastewater treatment facilities and subsequently to agricultural soils via application of contaminated sludge. On-route, the chemical properties of Ag may change, and further alterations are possible upon entry to soil. In the present study, we examined the long-term stability and (bio)availability of Ag along the "wastewater-sludge-soil" pathway. Synchrotron-based X-ray absorption spectroscopy (XAS) revealed that ca. 99% of Ag added to the sludge reactors as either Ag-NPs or AgNO3 was retained in sludge, with ≥79% of this being transformed to Ag2S, with the majority (≥87%) remaining in this form even after introduction to soils at various pH values and Cl concentrations for up to 400 days. Diffusive gradients in thin films (DGT), chemical extraction, and plant uptake experiments indicated that the potential (bio)availability of Ag in soil was low but increased markedly in soils with elevated Cl, likely due to the formation of soluble AgClx complexes in the soil solution. Although high Cl concentrations increased the bioavailability of Ag markedly, plant growth was not reduced in any treatment. Our results indicate that Ag-NPs entering soils through the wastewater-sludge-soil pathway pose low risk to plants due to their conversion to Ag2S in the wastewater treatment process, although bioavailability may increase in saline soils or when irrigated with high-Cl water.

Nanomaterials in Biosolids Inhibit Nodulation, Shift Microbial Community Composition, and Result in Increased Metal Uptake Relative to Bulk/Dissolved Metals.

We examined the effects of amending soil with biosolids produced from a pilot-scale wastewater treatment plant containing a mixture of metal-based engineered nanomaterials (ENMs) on the growth of Medicago truncatula, its symbiosis with Sinorhizobium meliloti, and on soil microbial community structure. Treatments consisted of soils amended with biosolids generated with (1) Ag, ZnO, and TiO2 ENMs introduced into the influent wastewater (ENM biosolids), (2) AgNO3, Zn(SO4)2, and micron-sized TiO2 (dissolved/bulk metal biosolids) introduced into the influent wastewater stream, or (3) no metal added to influent wastewater (control). Soils were amended with biosolids to simulate 20 years of metal loading, which resulted in nominal metal concentrations of 1450, 100, and 2400 mg kg(-1) of Zn, Ag, and Ti, respectively, in the dissolved/bulk and ENM treatments. Tissue Zn concentrations were significantly higher in the plants grown in the ENM treatment (182 mg kg(-1)) compared to those from the bulk treatment (103 mg kg(-1)). Large reductions in nodulation frequency, plant growth, and significant shifts in soil microbial community composition were found for the ENM treatment compared to the bulk/dissolved metal treatment. These results suggest differences in metal bioavailability and toxicity between ENMs and bulk/dissolved metals at concentrations relevant to regulatory limits.

Multitechnique investigation of the pH dependence of phosphate induced transformations of ZnO nanoparticles.

In order to properly evaluate the ecological and human health risks of ZnO manufactured nanomaterials (MNMs) released to the environment, it is critical to understand the likely transformation products in various environments, such as soils, surface and ground waters, and wastewater treatment processes. To address this knowledge gap, we examined the transformation of 30 nm ZnO MNMs in the presence of different concentrations of phosphate as a function of time and pH using a variety of orthogonal analytical techniques. The data reveal that ZnO MNMs react with phosphate at various concentrations and transform into two distinct morphological/structural phases: a micrometer scale crystalline zinc phosphate phase (hopeite-like) and a nanoscale phase that likely consists of a ZnO core with an amorphous Zn3(PO4)2 shell. The P species composition was also pH dependent, with 82% occurring as hopeite-like P at pH 6 while only 15% occurred as hopeite-like P at pH 8. These results highlight how reactions of ZnO MNMs with phosphate are influenced by environmental variables, including pH, and may ultimately result in structurally and morphologically heterogeneous end products.

The Importance of Fostering and Funding Scientific Research, and its Relevance to Environmental Toxicology and Chemistry.

What do environmental contaminants and climate change have in common with the virus SARS-CoV-2 and the disease COVID-19? We argue that one common element is the wealth of basic and applied scientific research that provides the knowledge and tools essential in developing effective programs for addressing threats to humans and social-ecological systems. Research on various chemicals, including dichlorodiphenyltrichloroethane and per- and polyfluoroalkyl substances, resulted in regulatory action to protect environmental and human health. Moreover, decades of research on coronaviruses, mRNA, and recently SARS-CoV-2 enabled the rapid development of vaccines to fight the COVID-19 pandemic. In the present study, we explore the common elements of basic and applied scientific research breakthroughs that link chemicals, climate change, and SARS-CoV-2/COVID-19 and describe how scientific information was applied for protecting human health and, more broadly, socio-ecological systems. We also offer a cautionary note on the misuse and mistrust of science that is not new in human history, but unfortunately is surging in modern times. Our goal was to illustrate the critical role of scientific research to society, and we argue that research must be intentionally fostered, better funded, and applied appropriately. To that end, we offer evidence that supports the importance of investing in scientific research and, where needed, ways to counter the spread of misinformation and disinformation that undermines legitimate discourse. Environ Toxicol Chem 2023;42:581-593. © 2022 SETAC.

Bioaccumulation of gold nanomaterials by Manduca sexta through dietary uptake of surface contaminated plant tissue.

We investigated the potential for bioaccumulation of engineered nanomaterials (ENMs) by tobacco hornworm (Manduca sexta) caterpillars resulting from the ingestion of plant tissue surface contaminated with ENMs. Caterpillars were fed tomato leaf tissue that had been surface contaminated with 12 nm tannate coated Au ENMs. After dosing was complete, bulk Au concentrations in individual caterpillars were measured after 0, 1, 4, and 7 days of elimination. Growth, mortality, and ingestion rate were monitored. This experiment revealed (1) no evidence that caterpillars were affected by ingestion of ENM contaminated plant tissue; (2) low bioaccumulation factors (BAF = 0.16) compared to a previous study where hornworm caterpillars were fed plants that had previously bioaccumulated Au ENMs (BAF = 6.2-11.6); (3) inefficient elimination of accumulated Au ENMs not associated with hornworm gut contents; and (4) regional differences in translocation of Au ENMs into tissues surrounding the hornworm gut, possibly the result of the interaction between ENM surface chemistry and regional differences in hornworm gut chemistry. These data, along with previous findings, indicate that although ENMs resuspended from soil onto plant surfaces by wind, water, biota, and/or mechanical disturbances are bioavailable to terrestrial consumers, bioaccumulation efficiency may be much lower via this pathway than through direct trophic exposure.

Trophic transfer of Au nanoparticles from soil along a simulated terrestrial food chain.

To determine if nanoparticles (NPs) could be transferred from soil media to invertebrates and then to secondary consumers, we examined the trophic transfer of Au NPs along a simulated terrestrial food chain. Earthworms (Eisenia fetida) were exposed to Au NPs in artificial soil media and fed to juvenile bullfrogs (Rana catesbeina). Earthworm Au concentrations were continuously monitored so that the cumulative dose to bullfrogs could be accurately estimated throughout the experiment. We exposed a second group of bullfrogs to equivalent doses of Au NPs by oral gavage to compare the bioavailability of NPs through direct exposure to trophic exposure. We observed accumulation of Au in liver, kidney, spleen, muscle, stomach, and intestine in both treatment groups. Tissue concentrations decreased on average of approximately 100-fold with each trophic-step. The total assimilated dose averaged only 0.09% of the administered dose for direct exposure (oral gavage), but 0.12% for the trophic exposure. The results suggest that manufactured NPs present in soil may be taken up into food chains and transferred to higher order consumers. They also suggest that Au NPs may be more bioavailable through trophic exposure than direct exposure and that trophic transfer may influence the biodistribution of particles once absorbed.

Bioavailability of gold nanomaterials to plants: importance of particle size and surface coating.

We used the model organisms Nicotiana tabacum L. cv Xanthi (tobacco) and Triticum aestivum (wheat) to investigate plant uptake of 10-, 30-, and 50-nm diameter Au manufactured nanomaterials (MNMs) coated with either tannate (T-MNMs) or citrate (C-MNMs). Primary particle size, hydrodynamic size, and zeta potential were characterized using transmission electron microscopy (TEM), dynamic light scattering (DLS), and electrophoretic mobility measurements, respectively. Plants were exposed to NPs hydroponically for 3 or 7 days for wheat and tobacco, respectively. Volume averaged Au concentrations were determined using inductively coupled plasma mass spectrometry (ICP-MS). Spatial distribution of Au in tissue samples was determined using laser ablation ICP-MS (LA-ICP-MS) and scanning X-ray fluorescence microscopy (µXRF). Both C-MNMs and T-MNMs of each size treatment bioaccumulated in tobacco, but no bioaccumulation of MNMs was observed for any treatment in wheat. These results indicate that MNMs of a wide range of size and with different surface chemistries are bioavailable to plants, provide mechanistic information regarding the role of cell wall pores in plant uptake of MNMs, and raise questions about the importance of plant species to MNM bioaccumulation.

Toxicogenomic responses of the model organism Caenorhabditis elegans to gold nanoparticles.

We used Au nanoparticles (Au-NPs) as a model for studying particle-specific effects of manufactured nanomaterials (MNMs) by examining the toxicogenomic responses in a model soil organism, Caenorhabditis elegans . Global genome expression for nematodes exposed to 4-nm citrate-coated Au-NPs at the LC(10) level (5.9 mg·L(-1)) revealed significant differential expression of 797 genes. The levels of expression for five genes (apl-1, dyn-1, act-5, abu-11, and hsp-4) were confirmed independently with qRT-PCR. Seven common biological pathways associated with 38 of these genes were identified. Up-regulation of 26 pqn/abu genes from noncanonical unfolded protein response (UPR) pathway and molecular chaperones (hsp-16.1, hsp-70, hsp-3, and hsp-4) were observed and are likely indicative of endoplasmic reticulum stress. Significant increase in sensitivity to Au-NPs in a mutant from noncanonical UPR (pqn-5) suggests possible involvement of the genes from this pathway in a protective mechanism against Au-NPs. Significant responses to Au-NPs in endocytosis mutants (chc-1 and rme-2) provide evidence for endocytosis pathway being induced by Au-NPs. These results demonstrate that Au-NPs are bioavailable and cause adverse effects to C. elegans by activating both general and specific biological pathways. The experiments with mutants further support involvement of several of these pathways in Au-NP toxicity and/or detoxification.

Wastewater surveillance demonstrates high predictive value for COVID-19 infection on board repatriation flights to Australia.

Controlling importation and transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) from overseas travelers is essential for countries, such as Australia, New Zealand, and other island nations, that have adopted a suppression strategy to manage very low community transmission. Wastewater surveillance of SARS-CoV-2 RNA has emerged as a promising tool employed in public health response in many countries globally. This study aimed to establish whether the surveillance of aircraft wastewater can be used to provide an additional layer of information to augment individual clinical testing. Wastewater from 37 long-haul flights chartered to repatriate Australians was tested for the presence of SARS-CoV-2 RNA. Children 5 years or older on these flights tested negative for coronavirus disease 19 (COVID-19) (deep nasal and oropharyngeal reverse-transcription (RT)-PCR swab) 48 h before departure. All passengers underwent mandatory quarantine for 14-day post arrival in Howard Springs, NT, Australia. Wastewater from 24 (64.9 %) of the 37 flights tested positive for SARS-CoV-2 RNA. During the 14 day mandatory quarantine, clinical testing identified 112 cases of COVID-19. Surveillance for SARS-CoV-2 RNA in repatriation flight wastewater using pooled results from three RT-qPCR assays demonstrated a positive predictive value (PPV) of 87.5 %, a negative predictive value (NPV) of 76.9 % and 83.7% accuracy for COVID-19 cases during the post-arrival 14-day quarantine period. The study successfully demonstrates that the surveillance of wastewater from aircraft for SARS-CoV-2 can provide an additional and effective tool for informing the management of returning overseas travelers and for monitoring the importation of SARS CoV-2 and other clinically significant pathogens.

Detection of the Omicron (B.1.1.529) variant of SARS-CoV-2 in aircraft wastewater.

On the 26th of November 2021, the World Health Organization (WHO) designated the newly detected B.1.1.529 lineage of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) the Omicron Variant of Concern (VOC). The genome of the Omicron VOC contains more than 50 mutations, many of which have been associated with increased transmissibility, differing disease severity, and potential to evade immune responses developed for previous VOCs such as Alpha and Delta. In the days since the designation of B.1.1.529 as a VOC, infections with the lineage have been reported in countries around the globe and many countries have implemented travel restrictions and increased border controls in response. We putatively detected the Omicron variant in an aircraft wastewater sample from a flight arriving to Darwin, Australia from Johannesburg, South Africa on the 25th of November 2021 via positive results on the CDC N1, CDC N2, and del(69-70) RT-qPCR assays per guidance from the WHO. The Australian Northern Territory Health Department detected one passenger onboard the flight who was infected with SARS-CoV-2, which was determined to be the Omicron VOC by sequencing of a nasopharyngeal swab sample. Subsequent sequencing of the aircraft wastewater sample using the ARTIC V3 protocol with Nanopore and ATOPlex confirmed the presence of the Omicron variant with a consensus genome that clustered with the B.1.1.529 BA.1 sub-lineage. Our detection and confirmation of a single onboard Omicron infection via aircraft wastewater further bolsters the important role that aircraft wastewater can play as an independent and unintrusive surveillance point for infectious diseases, particularly coronavirus disease 2019.

Minimizing errors in RT-PCR detection and quantification of SARS-CoV-2 RNA for wastewater surveillance.

Wastewater surveillance for pathogens using reverse transcription-polymerase chain reaction (RT-PCR) is an effective and resource-efficient tool for gathering community-level public health information, including the incidence of coronavirus disease-19 (COVID-19). Surveillance of Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) in wastewater can potentially provide an early warning signal of COVID-19 infections in a community. The capacity of the world's environmental microbiology and virology laboratories for SARS-CoV-2 RNA characterization in wastewater is increasing rapidly. However, there are no standardized protocols or harmonized quality assurance and quality control (QA/QC) procedures for SARS-CoV-2 wastewater surveillance. This paper is a technical review of factors that can cause false-positive and false-negative errors in the surveillance of SARS-CoV-2 RNA in wastewater, culminating in recommended strategies that can be implemented to identify and mitigate some of these errors. Recommendations include stringent QA/QC measures, representative sampling approaches, effective virus concentration and efficient RNA extraction, PCR inhibition assessment, inclusion of sample processing controls, and considerations for RT-PCR assay selection and data interpretation. Clear data interpretation guidelines (e.g., determination of positive and negative samples) are critical, particularly when the incidence of SARS-CoV-2 in wastewater is low. Corrective and confirmatory actions must be in place for inconclusive results or results diverging from current trends (e.g., initial onset or reemergence of COVID-19 in a community). It is also prudent to perform interlaboratory comparisons to ensure results' reliability and interpretability for prospective and retrospective analyses. The strategies that are recommended in this review aim to improve SARS-CoV-2 characterization and detection for wastewater surveillance applications. A silver lining of the COVID-19 pandemic is that the efficacy of wastewater surveillance continues to be demonstrated during this global crisis. In the future, wastewater should also play an important role in the surveillance of a range of other communicable diseases.

Evidence for biomagnification of gold nanoparticles within a terrestrial food chain.

Nanoparticles from the rapidly increasing number of consumer products that contain manufactured nanomaterials are being discharged into waste streams. Increasing evidence suggests that several classes of nanomaterials may accumulate in sludge derived from wastewater treatment and ultimately in soil following land application as biosolids. Little research has been conducted to evaluate the impact of nanoparticles on terrestrial ecosystems, despite the fact that land application of biosolids from wastewater treatment will be a major pathway for the introduction of manufactured nanomaterials to the environment. To begin addressing this knowledge gap, we used the model organisms Nicotiana tabacum L. cv Xanthi and Manduca sexta (tobacco hornworm) to investigate plant uptake and the potential for trophic transfer of 5, 10, and 15 nm diameter gold (Au) nanoparticles (NPs). Samples were analyzed using both bulk analysis by inductively coupled plasma mass spectrometry (ICP-MS) as well as spatially resolved methods such as laser ablation inductively coupled mass spectrometry (LA-ICP-MS) and X-ray fluorescence (µXRF). Our results demonstrate trophic transfer and biomagnification of gold nanoparticles from a primary producer to a primary consumer by mean factors of 6.2, 11.6, and 9.6 for the 5, 10, and 15 nm treatments, respectively. This result has important implications for risks associated with nanotechnology, including the potential for human exposure.

Evidence for avoidance of Ag nanoparticles by earthworms (Eisenia fetida).

Silver nanoparticles have been incorporated into a wide variety of consumer products, ideally acting as antimicrobial agents. Silver exposure has long been known to cause toxic effects to a wide variety of organisms, making large scale production of silver nanoparticles a potential hazard to environmental systems. Here we describe the first evidence that an organism may be able to sense manufactured nanoparticles in a complex, environmentally relevant exposure and that the presence of nanoparticles alters the organism's behavior. We found that earthworms (Eisenia fetida) consistently avoid soils containing silver nanoparticles and AgNO(3) at similar concentrations of Ag. However, avoidance of silver nanoparticles occurred over 48 h, while avoidance of AgNO(3) was immediate. It was determined that avoidance of silver nanoparticles could not be explained by release of silver ions or any changes in microbial communities caused by the introduction of Ag. This leads us to conclude that the earthworms were in some way sensing the presence of nanoparticles over the course of a 48 h exposure and choosing to avoid exposure to them. Our results demonstrate that nanoparticle interactions with organisms may be unpredictable and that these interactions may result in ecologically significant effects on behavior at environmentally relevant concentrations.

SARS-CoV-2 RNA monitoring in wastewater as a potential early warning system for COVID-19 transmission in the community: A temporal case study.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the virus which causes coronavirus disease (COVID-19), has spread rapidly across the globe infecting millions of people and causing significant health and economic impacts. Authorities are exploring complimentary approaches to monitor this infectious disease at the community level. Wastewater-based epidemiology (WBE) approaches to detect SARS-CoV-2 RNA in municipal wastewater are being implemented worldwide as an environmental surveillance approach to inform health authority decision-making. Owing to the extended excretion of SARS-CoV-2 RNA in stool, WBE can surveil large populated areas with a longer detection window providing unique information on the presence of pre-symptomatic and asymptomatic cases that are unlikely to be screened by clinical testing. Herein, we analysed SARS-CoV-2 RNA in 24-h composite wastewater samples (n = 63) from three wastewater treatment plants (WWTPs) in Brisbane, Queensland, Australia from 24th of February to 1st of May 2020. A total of 21 samples were positive for SARS-CoV-2, ranging from 135 to 11,992 gene copies (GC)/100 mL of wastewater. Detections were made in a Southern Brisbane WWTP in late February 2020, up to three weeks before the first clininal case was reported there. Wastewater samples were generally positive during the period with highest caseload data. The positive SARS-CoV-2 RNA detection in wastewater while there were limited clinical reported cases demonstrates the potential of WBE as an early warning system to identify hotspots and target localised public health responses, such as increased individual testing and the provision of health warnings.

Evidence for bioavailability of Au nanoparticles from soil and biodistribution within earthworms (Eisenia fetida).

Because Au nanoparticles (NPs) are resistant to oxidative dissolution and are easily detected, they have been used as stable probes for the behavior of nanomaterials within biological systems. Previous studies provide somewhat limited evidence for bioavailability of Au NPs in food webs, because the spatial distribution within tissues and the speciation of Au was not determined. In this study, we provide multiple lines of evidence, including orthogonal microspectroscopic techniques, as well as evidence from biological responses, that Au NPs are bioavailable from soil to a model detritivore (Eisenia fetida). We also present limited evidence that Au NPs may cause adverse effects on earthworm reproduction. This is perhaps the first study to demonstrate that Au NPs can be taken up by detritivores from soil and distributed among tissues. We found that primary particle size (20 or 55 nm) did not consistently influence accumulated concentrations on a mass concentration basis; however, on a particle number basis the 20 nm particles were more bioavailable. Differences in bioavailability between the treatments may have been explained by aggregation behavior in pore water. The results suggest that nanoparticles present in soil from activities such as biosolids application have the potential to enter terrestrial food webs.

Effects of particle size on chemical speciation and bioavailability of copper to earthworms (Eisenia fetida) exposed to copper nanoparticles.

To investigate the role of particle size on the oxidation, bioavailability, and adverse effects of manufactured Cu nanoparticles (NPs) in soils, we exposed the earthworm Eisenia ferida to a series of concentrations of commercially produced NPs labeled as 20- to 40-nm or < 100-nm Cu in artificial soil media. Effects on growth, mortality, reproduction, and expression of a variety of genes associated with metal homeostasis, general stress, and oxidative stress were measured. We also used X-ray absorption spectroscopy and scanning X-ray fluorescence microscopy to characterize changes in chemical speciation and spatial distribution of the NPs in soil media and earthworm tissues. Exposure concentrations of Cu NPs up to 65 mg kg(-1) caused no adverse effects on ecologically relevant endpoints. Increases in metallothionein expression occurred at concentrations exceeding 20 mg kg(-1) of Cu NPs and concentrations exceeding 10 mg kg(-1) of CuSO4. Based on the relationship of Cu tissue concentration to metallothionein expression level and the spatial distribution and chemical speciation of Cu in the tissues, we conclude that Cu ions and oxidized Cu NPs were taken up by the earthworms. This study suggests that oxidized Cu NPs may enter food chains from soil but that adverse effects in earthworms are likely to occur only at relatively high concentrations (> 65 mg Cu kg(-1) soil).

Application of sewage sludge containing environmentally-relevant silver sulfide nanoparticles increases emissions of nitrous oxide in saline soils.

Silver (Ag) is released from a range of products and accumulates in agricultural soils as silver sulfide (Ag2S) through the application of Ag-containing biosolids as a soil amendment. Although Ag2S is comparatively stable, its solubility increases with salinity, potentially altering its impacts on microbial communities due to the anti-microbial properties of Ag. In this study, we investigated the impacts of Ag on the microbially mediated N cycle in saline soils by examining the relationship between the (bio)availability of Ag2S and microbial functioning following the application of Ag2S-containing sludge. Synchrotron-based X-ray absorption spectroscopy (XAS) revealed that the Ag2S was stable within the soil, although extractable Ag concentrations increased up to 18-fold in soils with higher salinity. However, the extractable Ag accounted for <0.05% of the total Ag in all soils and had no impact on plant biomass or soil bacterial biomass. Interestingly, at high soil salinity, Ag2S significantly increased cumulative N2O emissions from 80.9 to 229.2 mg kg-1 dry soil (by 180%) compared to the corresponding control sludge treatment, which was ascribed to the increased abundance of nitrification and denitrification-related genes (amoA, nxrB, narG, napA, nirS, and nosZ) and increased relative abundance of denitrifiers (Rhodanobacter, Salinimicrobium, and Zunongwangia). Together, our findings show that the application of Ag2S-containing sludge to a saline soil can disrupt the N cycle and increase N2O emissions from agroecosystems.