Arsenic, cadmium, lead, antimony bioaccessibility and relative bioavailability in legacy gold mining waste.

Bioaccessibility and relative bioavailability of As, Cd, Pb and Sb was investigated in 30 legacy gold mining wastes (calcine sands, grey battery sands, tailings) from Victorian goldfields (Australia). Pseudo-total As concentration in 29 samples was 1.45-148-fold higher than the residential soil guidance value (100 mg/kg) while Cd and Pb concentrations in calcine sands were up to 2.4-fold and 30.1-fold higher than the corresponding guidance value (Cd: 20 mg/kg and Pb: 300 mg/kg). Five calcine sands exhibited elevated Sb (31.9-5983 mg/kg), although an Australian soil guidance value is currently unavailable. Arsenic bioaccessibility (n = 30) and relative bioavailability (RBA; n = 8) ranged from 6.10-77.6% and 10.3-52.9% respectively. Samples containing > 50% arsenopyrite/scorodite showed low As bioaccessibility (<20.0%) and RBA (<15.0%). Co-contaminant RBA was assessed in 4 calcine sands; Pb RBA ranged from 73.7-119% with high Pb RBA associated with organic and mineral sorbed Pb and, lower Pb RBA observed in samples containing plumbojarosite. In contrast, Cd RBA ranged from 55.0-67.0%, while Sb RBA was < 5%. This study highlights the importance of using multiple lines of evidence during exposure assessment and provides valuable baseline data for co-contaminants associated with legacy gold mining activities.

Impact of precursors and bioaccessibility on childhood PFAS exposure from house dust.

This study investigated the impact of precursors and bioaccessibility on childhood per- and polyfluoroalkyl substances (PFAS) exposure in house dust (n = 28) from Adelaide, Australia. Sum PFAS concentration (∑38) ranged from 3.0 to 2640 µg kg-1 with PFOS (1.5-675 µg kg-1), PFHxS (1.0-405 µg kg-1) and PFOA (1.0-155 µg kg-1) constituting the major perfluoroalkyl sulfonic (PFSA) and carboxylic acids (PFCA). The total oxidizable precursor (TOP) assay was applied to estimate concentrations of unmeasurable precursors that may undergo oxidation to measurable PFAS. Sum PFAS concentration post-TOP assay changed 3.8-112-fold (91.5-62,300 µg kg-1) with median post-TOP assay PFCA (C4-C8) concentrations (92.3-170 µg kg-1) increasing significantly (13.7-48.5-fold). As incidental dust ingestion is a significant exposure pathway for young children, PFAS bioaccessibility was determined using an in vitro assay. Sum PFAS bioaccessibility ranged from 4.6 to 49.3 % with significantly (p < 0.05) higher PFCA (10.3-83.4 %) bioaccessibility compared to PFSA (3.5-51.5 %). When in vitro extracts were assessed post-TOP assay, PFAS bioaccessibility changed (7-1060 versus 137-3900 µg kg-1) although percentage bioaccessibility decreased (2.3-14.5 %) due to the disproportionately higher post-TOP assay PFAS concentration. PFAS estimated daily intake (EDI) was calculated for a 'stay-at-home' 2-3-year-old child. Inclusion of dust specific bioaccessibility values resulted in a 1.7-20.5-fold decrease in ∑PFOA, PFOA and PFHxS EDI (0.02-1.23 ng kg bw-1 day-1) compared to default absorption assumptions (0.23-5.4 ng kg bw-1 day-1). However, when 'worst-case scenario' precursor transformation was considered, EDI calculations were 4.1-187-fold higher that the EFSA tolerable weekly intake value (equivalent to 0.63 ng kg bw-1 day-1), although this was moderated when exposure parameters were refined through PFAS bioaccessibility incorporation (0.35-17.0-fold higher than the TDI). Irrespective of exposure scenario, EDI calculations were below the FSANZ tolerable daily intake values for PFOS (20 ng kg bw-1 day-1) and PFOA (160 ng kg bw-1 day-1) for all dust samples analysed.

Metallic mangroves: Sediments and in situ diffusive gradients in thin films (DGTs) reveal Avicennia marina (Forssk.) Vierh. lives with high contamination near a lead­zinc smelter in South Australia.

From 1889, aerial emissions and effluent from a coastal lead­zinc smelter at Port Pirie, South Australia, have led to the accumulation of lead (Pb), zinc (Zn), arsenic (As), cadmium (Cd) and copper (Cu) in the surrounding marine environment. Despite this, extensive stands of grey mangrove (Avicennia marina) inhabit coastal areas at Port Pirie, right up to the smelter's boundary. To understand the contamination level the mangroves are living in there, elemental concentrations were measured in mangrove sediments, leaves, pneumatophores and fruits at sites 0.30-43.0 km from the smelter. Plant health was assessed via leaf chlorophyll content at four sites with contrasting contamination, as well as in situ labile elemental concentration using diffusive gradients in thin films (DGT). Sites < 1.7 km of the smelter exceeded Australian and New Zealand Environment and Conservation Council (ANZECC) & Agriculture and Resource Management Council of Australia and New Zealand (ARMCANZ) (2000) sediment quality guideline values for As (78.3-191 mg/kg), Cd (5.17-151 mg/kg), Cu (80.7-788 mg/kg), Pb (2,544-14,488 mg/kg) and Zn (281-62,097 mg/kg), while sites further away showed less enrichment above background. Similarly, elevated elemental concentrations in leaves and pneumatophores occurred closer to the smelter (up to 319 mg/kg Pb; 1,033 mg/kg Zn), while fruits had little contamination of non-essential elements (≤ 5.23 mg/kg). Relationship between sediment and leaf elemental concentration was isometric for Pb and anisometric for others. Labile As, Cd, Cu, Pb and Zn exceeded the 95% and 80% level of species protection in marine water by ANZECC & ARMCANZ (2000) near the smelter, but chlorophyll content did not vary significantly among sites (p > 0.05). These results reveal that A. marina tolerate high elemental contamination at Port Pirie, contributing to lesser but still high contamination in plants, warranting further investigation into non-lethal impacts on mangroves or additional biota inhabiting this ecosystem.

Application of soil amendments for reducing PFAS leachability and bioavailability.

In this study, changes in PFAS leachability and bioavailability were determined following the application of RemBind®100 (R100) and RemBind®300 (R300; 1-10% w/w) to PFAS-contaminated soil (Æ©28 PFAS 3.093-32.78 mg kg-1). Small differences were observed in PFAS immobilization efficacy when soil was amended with RemBind® products although adding 5% w/w of either product resulted in a >98% reduction in ASLP PFAS leachability. Variability in immobilization efficacy was attributed to differences in activated carbon composition which influenced physicochemical properties of RemBind® formulations and PFAS sorption. PFOS, PFHxS and PFOA relative bioavailability was also assessed in unamended and amended soil (5% w/w) using an in vivo mouse model. In unamended soil, PFAS relative bioavailability was >60% with differences attributed to physicochemical properties of soil which influenced electrostatic and hydrophobic interactions. However, when PFAS relative bioavailability was assessed in soil amended with 5% w/w R100, individual PFAS relative bioavailability was reduced to 16.1 ± 0.8% to 26.1 ± 0.9% with similar results observed when R300 (5% w/w) was utilised (14.4 ± 1.6% to 24.3 ± 0.8%). Results from this study highlight that soil amendments have the potential to reduce both PFAS leachability and relative bioavailability thereby decreasing mobility and potential exposure to soil-borne contaminants.

Plumbojarosite formation in contaminated soil to mitigate childhood exposure to lead, arsenic and antimony.

In this study, a novel method for lead (Pb) immobilization was developed in contaminated soils using iron (III) (Fe3+) in conjunction with 0.05 M H2SO4. During method optimization, a range of microwave treatment times, solid to solution ratios, and Fe2(SO4)3/H2SO4 concentrations were assessed using a mining/smelting impacted soil (BHK2, Pb: 3031 mg/kg), followed by treatment of additional Pb contaminated soils (PP, Pb: 1506 mg/kg, G10, Pb: 2454 mg/kg and SoFC-1, Pb: 6340 mg/kg) using the optimized method. Pb bioaccessibility was assessed using USEPA Method 1340, with Pb speciation determined by X-ray Absorption (XAS) spectroscopy. Treatment efficacy was also validated using an in vivo mouse assay, where Pb accumulation in femur, kidney and liver was assessed to confirm in vitro bioaccessibility outcomes. Results showed that Pb bioaccessibility could be reduced by 77.4-97.0% following treatment of soil with Fe2(SO4)3 (0.4-1.0 M), H2SO4 (0.05 M) at 150 °C for 60 min in a closed microwave system. Results of bioavailability assessment demonstrated treatment effect ratio of 0.06-0.07 in femur, 0.06-0.27 in kidney and 0.06-0.11 in liver (bioavailability reduction between 73% and 93%). Formation of plumbojarosite in treated soils was confirmed by XAS analysis.

Plastic, nutrition and pollution; relationships between ingested plastic and metal concentrations in the livers of two Pachyptila seabirds.

Naturally occurring metals and metalloids [metal(loid)s] are essential for the physiological functioning of wildlife; however, environmental contamination by metal(loid) and plastic pollutants is a health hazard. Metal(loid)s may interact with plastic in the environment and there is mixed evidence about whether plastic ingested by wildlife affects metal(loid) absorption/assimilation and concentration in the body. We examined ingested plastic and liver concentration of eleven metal(loid)s in two seabird species: fairy (Pachyptila turtur) and slender-billed prions (P. belcheri). We found significant relationships between ingested plastic and the concentrations of aluminium (Al), manganese (Mn), iron (Fe), cobalt (Co), copper (Cu) and zinc (Zn) in the liver of prions. We investigated whether the pattern of significant relationships reflected plastic-metal(loid) associations predicted in the scientific literature, including by transfer of metals from ingested plastics or malnutrition due to dietary dilution by plastics in the gut. We found some support for both associations, suggesting that ingested plastic may be connected with dietary dilution / lack of essential nutrients, especially iron, and potential transfer of zinc. We did not find a relationship between plastic and non-essential metal(loid)s, including lead. The effect of plastic was minor compared to that of dietary exposure to metal(oid)s, and small plastic loads (< 3 items) had no discernible link with metal(loid)s. This new evidence shows a relationship between plastic ingestion and liver metal(loid) concentrations in free-living wildlife.

Influence of household smoking habits on inhalation bioaccessibility of trace elements and light rare earth elements in Canadian house dust.

In this study, total concentration and inhalation bioaccessibility (dissolution in simulated biological solution) of trace elements (TE) and rare earth elements (REE) were assessed in PM10 from Canadian house dust samples with smoking (n = 25) and non-smoking (n = 25) status. Compared to the natural background concentrations in Canadian soils, median Zn, Pb, Cd and Cu concentrations in PM10 were 10-23 fold higher, while median La, Ce and Pr concentrations were 1.6-2.4 fold higher. Mann-Whitney tests (α = 0.05) indicated no difference between the median TE concentrations based on the smoking status of the household; however, median REE concentrations were significantly higher in the PM10 of smoking households. Additionally, Cd and Ni were positively correlated (Spearman r, p < 0.05) to La, Ce and Nd in smoking households, suggesting that tobacco combustion may have contributed REE in the PM10 of these households. Median inhalation-ingestion bioaccessibility assay outcomes of arsenic (As) and lead (Pb) was higher in the non-smoking households when compared to smoking households (Mann Whitney test, α = 0.05), suggesting that tobacco combustion products may be associated with less soluble species of As and Pb. Although REE bioaccessibility was negligible in simulated lung epithelial fluid regardless of the smoking status of the household, bioaccessibility in the lung-gastric phase was 23.6-27.6% in the smoking household and 34.7-36.7% in the non-smoking households, indicating a significantly lower REE dissolution in PM10 of smoking households. In contrast, between 17 and 21.9% bioaccessibility of REE was observed when artificial lysosomal fluid was used, where the outcome was not significantly affected by the smoking status. This study indicates that despite a higher median REE concentration in the PM10 of smoking households, inhalation bioaccessibility may be significantly influenced by the mineralogy.

Correlation between lead speciation and inhalation bioaccessibility using two different simulated lung fluids.

This study investigated the relationship between lead (Pb) speciation determined using Extended X-ray Absorption Fine Structure (EXAFS) spectroscopy in <10 µm particulate matter (PM10) from mining/smelting impacted Australian soils (PP, BHK5, BHK6, BHK10 and BHK11) and inhalation exposure using two simulated lung fluids [Hatch's solution, pH 7.4 and artificial lysosomal fluid (ALF), pH 4.5]. Additionally, elemental composition of Pb rich regions in PP PM10 and the post-bioaccessibility assay residuals were assessed using a combination of Scanning Electron Microscopy (SEM) and Energy Dispersive X-ray Spectroscopy (EDX) to provide insights into how extraction using simulated lung fluids may influence Pb speciation in vitro. Correlation between Pb speciation (weighted %) and bioaccessibility (%) was assessed using Pearson r (α = 0.1 and 0.05). Lead concentration in PM10 samples ranged from 782 mg/kg (BHK6) to 7796 mg/kg (PP). Results of EXAFS analysis revealed that PP PM10 was dominated by Pb adsorbed onto clay/oxide, while the four BHK PM10 samples showed variability in the weighted % of Pb adsorbed onto clay/oxide and organic matter bound Pb, Pb phosphate, anglesite and galena. When bioaccessibility was assessed using different in vitro inhalation assays, results varied between samples and between assays, Pb bioaccessibility in Hatch's solution ranged from 24.4 to 48.4%, while in ALF, values were significantly higher (72.9-96.3%; p < 0.05). When using Hatch's solution, bioaccessibility outcomes positively correlated to anglesite (r:0.6246, p:0.0361) and negatively correlated to Pb phosphate (r: -0.9610, p:0.0041), organic bound Pb (r: -0.7079, p: 0.0578), Pb phosphate + galena + plumbojarosite (r: -0.9350, p: 0.0099). No correlation was observed between Pb bioaccessibility (%) using Hatch's solution and weighted % of Pb adsorbed onto clay/oxide and between bioaccessibility (%) using ALF and any Pb species. SEM and EDX analysis revealed that a layer of O-Pb-Ca-P-Si-Al-Fe formed during the in vitro extraction using Hatch's solution.

Dynamics of Lead Bioavailability and Speciation in Indoor Dust and X-ray Spectroscopic Investigation of the Link between Ingestion and Inhalation Pathways.

Lead (Pb) exposure from household dust is a major childhood health concern because of its adverse impact on cognitive development. This study investigated the absorption kinetics of Pb from indoor dust following a single dose instillation into C57BL/6 mice. Blood Pb concentration (PbB) was assessed over 24 h, and the dynamics of particles in the lung and gastro-intestinal (GI) tract were visualized using X-ray fluorescence (XRF) microscopy. The influence of mineralogy on Pb absorption and particle retention was investigated using X-ray absorption near-edge structure spectroscopy. A rapid rise in PbB was observed between 0.25 and 4 h after instillation, peaking at 8 h and slowly declining during a period of 24 h. Following clearance from the lungs, Pb particles were detected in the stomach and small intestine at 4 and 8 h, respectively. Analysis of Pb mineralogy in the residual particles in tissues at 8 h showed that mineral-sorbed Pb and Pb-phosphates dominated the lung, while organic-bound Pb and galena were the main phases in the small intestines. This is the first study to visualize Pb dynamics in the lung and GI tract using XRF microscopy and link the inhalation and ingestion pathways for metal exposure assessment from dust.

Relationship between Pb relative bioavailability and bioaccessibility in phosphate amended soil: Uncertainty associated with predicting Pb immobilization efficacy using in vitro assays.

In this study, an in vitro in vivo correlation (IVIVC) between Pb in vitro bioaccessibility (IVBA) and relative bioavailability (RBA) was explored to determine whether the efficacy of Pb immobilization in phosphate amended soils could be predicted using an in vitro approach. Mining/smelting impacted soil from Broken Hill, Australia (582-3536 mg/kg of Pb in the <250 µm soil particle fraction) was amended with Phosphoric Acid (PA), Mono Ammonium Phosphate (MAP) or Triple Super Phosphate (TSP) at Pb:P molar ratios of 1:1-1:5. Pb speciation in pre- and post-treated soil was assessed using X-ray Absorption Spectroscopy (XAS), Pb IVBA was measured using the Solubility Bioaccessibility Research Consortium (SBRC) assay (gastric and intestinal phases), and Pb RBA was determined in mice using blood Pb concentration as the bioavailability endpoint. XAS analysis revealed a 3.75-6.00 fold increase in the weighted % of Pb phosphates in soil containing >1000 mg/kg Pb while treatment effect ratios of 0.89-0.99 (SBRC-G), 0.09-0.71 (SBRC-I) and 0.27-0.80 (RBA) were observed in PA amended soil (Pb:P = 1:5). Although significant (p < 0.05) correlation were obtained between Pb RBA and IVBA (%) determined using SBRC-G (r = 0.64) and SBRC-I (r = 0.67), the strengths of the relationships were weak (r2 = 0.41-0.45). This research highlights the complexities associated with the prediction of Pb RBA in phosphate amended soil.

In Vitro, in Vivo, and Spectroscopic Assessment of Lead Exposure Reduction via Ingestion and Inhalation Pathways Using Phosphate and Iron Amendments.

This study compared lead (Pb) immobilization efficacies in mining/smelting impacted soil using phosphate and iron amendments via ingestion and inhalation pathways using in vitro and in vivo assays, in conjunction with investigating the dynamics of dust particles in the lungs and gastro-intestinal tract via X-ray fluorescence (XRF) microscopy. Phosphate amendments [phosphoric acid (PA), hydroxyapatite, monoammonium phosphate (MAP), triple super phosphate (TSP), and bone meal biochar] and hematite were applied at a molar ratio of Pb:Fe/P = 1:5. Pb phosphate formation was investigated in the soil/post-in vitro bioaccessibility (IVBA) residuals and in mouse lung via extended X-ray absorption fine structure (EXAFS) and X-ray absorption near edge structures (XANES) spectroscopy, respectively. EXAFS analysis revealed that anglesite was the dominant phase in the ingestible (<250 µm) and inhalable (<10 µm) particle fractions. Pb IVBA was significantly reduced (p < 0.05) by phosphate amendments in the <250 µm fraction (solubility bioaccessibility research consortium assay) and by PA, MAP, and TSP in the <10 µm fraction (inhalation-ingestion bioaccessibility assay). A 21.1% reduction in Pb RBA (<250 µm fraction) and 56.4% reduction in blood Pb concentration (<10 µm fraction) were observed via the ingestion and inhalation pathways, respectively. XRF microscopy detected Pb in the stomach within 4 h, presumably via mucociliary clearance.

Methodological factors influencing inhalation bioaccessibility of metal(loid)s in PM2.5 using simulated lung fluid.

In this study, methodological factors influencing the dissolution of metal(loid)s in simulated lung fluid (SLF) was assessed in order to develop a standardised method for the assessment of inhalation bioaccessibility in PM2.5. To achieve this aim, the effects of solid to liquid (S/L) ratio (1:100 to 1:5000), agitation (magnetic agitation, occasional shaking, orbital and end-over-end rotation), composition of SLF (artificial lysosomal fluid: ALF; phagolysosomal simulant fluid: PSF) and extraction time (1-120 h) on metal(loid) bioaccessibility were investigated using PM2.5 from three Australian mining/smelting impacted soils and a certified reference material. The results highlighted that SLF composition significantly (p < 0.001) influenced metal(loid) bioaccessibility and that when a S/L ratio of 1:5000 and end-over-end rotation was used, metal(loid) solubility plateaued after approximately 24 h. Additionally, in order to assess the exposure of metal(loid)s via incidental ingestion of surface dust, PM2.5 was subjected to simulated gastro-intestinal tract (GIT) solutions and the results were compared to extraction using SLF. Although As bioaccessibility in SLF (24 h) was significantly lower than in simulated GIT solutions (p < 0.05), Pb bioaccessibility was equal to or significantly higher than that extracted using simulated GIT solutions (p < 0.05).

An inhalation-ingestion bioaccessibility assay (IIBA) for the assessment of exposure to metal(loid)s in PM10.

Although metal(loid) bioaccessibility of ambient particulate matter, with an aerodynamic diameter of <10µm (PM10), has recently received increasing attention, limited research exists into standardising in-vitro methodologies using simulated lung fluid (SLF). Contradictions exist regarding which assay parameters should be adopted. Additionally, potential continuation of metal(loid) dissolution once PM10 is cleared from the lungs and passed through the gastro-intestinal tract (GIT) has rarely been addressed. The objective of this study was to assess parameters that influence inhalation bioaccessibility in order to develop a conservative assay that is relevant to a human inhalation scenario. To achieve this aim, the effect of solid to liquid (S/L) ratio, extraction time, agitation and five major SLF compositions on the bioaccessibilities of arsenic (As) and lead (Pb) was investigated using PM10 from three Australian mining/smelting impacted regions. Using the biologically relevant parameters that resulted in the most conservative outcomes, bioaccessibility of metal(loid)s in PM10 was assessed in SLF, followed by simulated GIT solutions. Results from this study revealed that fluid composition and S/L ratio significantly affected metal(loid) dissolution (p<0.05). The highest Pb bioaccessibility resulted using simulated lung-gastric solution, while that of As resulted using simulated lung-gastric-small intestinal tract solutions. Compared to SLF alone, metal(loid) dissolution using the inhalation-ingestion bioaccessibility assay (IIBA) was significantly higher (p<0.05) for all PM10 samples.

A critical review of approaches and limitations of inhalation bioavailability and bioaccessibility of metal(loid)s from ambient particulate matter or dust.

Inhalation of metal(loid)s in ambient particulate matter (APM) represents a significant exposure pathway to humans. Although exposure assessment associated with this pathway is currently based on total metal(loid) content, a bioavailability (i.e. absorption in the systemic circulation) and/or bioaccessibility (i.e. solubility in simulated lung fluid) based approach may more accurately quantify exposure. Metal(loid) bioavailability-bioaccessibility assessment from APM is inherently complex and lacks consensus. This paper reviews the discrepancies that impede the adoption of a universal protocol for the assessment of inhalation bioaccessibility. Exposure assessment approaches for in-vivo bioavailability, in-vitro cell culture and in-vitro bioaccessibility (composition of simulated lungs fluid, physico-chemical and methodological considerations) are critiqued in the context of inhalation exposure refinement. An important limitation of bioavailability and bioaccessibility studies is the use of considerably higher than environmental metal(loid) concentration, which diminishing their relevance to human exposure scenarios. Similarly, individual metal(loid) studies have been criticised due to complexities of APM metal(loid) mixtures which may impart synergistic or antagonistic effects compared to single metal(loid) exposure. Although a number of different simulated lung fluid (SLF) compositions have been used in metal(loid) bioaccessibility studies, information regarding the comparative leaching efficiency among these different SLF and comparisons to in-vivo bioavailability data is lacking. In addition, the particle size utilised is often not representative of what is deposited in the lungs while assay parameters (extraction time, solid to liquid ratio, temperature and agitation) are often not biologically relevant. Research needs are identified in order to develop robust in-vitro bioaccessibility protocols for the assessment or prediction of metal(loid) bioavailability in APM for the refinement of inhalation exposure.

Ecotoxicity of neutral red (dye) and its environmental applications.

Neutral red (NR) is a synthetic phenazine with promising prospect in environmental biotechnology as an electron shuttle. Recently, NR injections into coal seam associated groundwater in Australia (final dissolved NR concentration: 8 µM ± 0.2) were shown to increase methanogenesis up to ten-fold. However, information about NR toxicity to ecological receptors is sorely lacking. The main aim of this study was to investigate the concentration dependent toxicity of NR in microorganisms and plants. Acute toxicity of NR was determined by the modified Microtox™ assay. Microbial viability was determined using Escherichia coli and Bacillus subtilis. Germination and early growth of plants was studied using Lactuca sativa, Daucus carota, Allium cepa and an Australian native Themeda triandra. Lastly, mutagenicity of the coal seam associated groundwater was assessed using the Ames test. The EC50 of acute NR toxicity was determined to be 0.11 mM. The EC50 of microbial viability was between 1 and 7.1mM NR. Among the concentrations tested, only 0.01, 0.10 and 100mM of NR significantly affected (p<0.001) germination of L. sativa. The EC50 for root elongation in seeds was between 1.2 and 35.5mM NR. Interestingly, root elongation in seeds was significantly stimulated (p<0.001) between 0.25 and 10mM NR, showing a hormetic effect. A significant increase in mutagenicity was only observed in one of the three wells tested. The results suggest that the average dissolved NR concentration (8 µM ± 0.2) deployed in the field trial at Lithgow State Coal Mine, Australia, appears not to negatively impact the ecological receptors tested in this study.