Arsenic speciation in the bracket fungus Fomitopsis betulina from contaminated and pristine sites.

Uptake, distribution and speciation of arsenic (As) were determined in the bracket fungus Fomitopsis betulina (previously Piptoporus betulinus), commonly known as the birch polypore, collected from a woodland adjacent to a highly contaminated former mine in the Southwest UK and at an uncontaminated site in Quebec, Canada, with no past or present mining activity. The fruiting body was divided into cap, centre and pores representing the top, middle and underside to identify trends in the distribution and transformation of As. Total As, determined by inductively coupled plasma-mass spectrometry (ICP-MS), was approximately tenfold higher in the mushroom from the contaminated compared to the uncontaminated site. Overall, accumulation of As was low relative to values reported for some soil-dwelling species, with maximum levels of 1.6 mg/kg at the contaminated site. Arsenic speciation was performed on aqueous extracts via both anion and cation high-performance liquid chromatography-ICP-MS (HPLC-ICP-MS) and on whole dried samples using X-ray absorption near edge structure (XANES) analysis. Seven As species were detected in F. betulina from the contaminated site by HPLC-ICP-MS: arsenite (AsIII), arsenate (AsV), dimethylarsinate (DMAV), methylarsonate (MAV), trimethylarsine oxide (TMAO), tetramethylarsonium ion (Tetra) and trace levels of arsenobetaine (AB). The same As species were observed at the uncontaminated site with the exception of TMAO and Tetra. Arsenic species were localized throughout the fruiting body at the contaminated site, with the cap and pores containing a majority of AsV, only the cap containing TMAO, and the pores containing higher concentrations of DMAV and MAV as well as tetra and a trace of AB. XANES analysis demonstrated that the predominant form of As at the contaminated site was inorganic AsIII coordinated with sulphur or oxygen and AsV coordinated with oxygen. This is the first account of arsenic speciation in F. betulina or any fungi of the family Fomitopsidaceae.

Tweets and Reactions: Revealing the Geographies of Cybercrime Perpetrators and the North-South Divide.

How do tweets reflect the long-standing disparities between the northern and southern regions of Nigeria? This study presents a qualitative analysis of Twitter users' responses (n = 101,518) to the tweets of the Economic and Financial Crimes Commission (EFCC) regarding the production and prosecution of cybercrime. The article uses postcolonial perspectives to shed light on the legacies of British colonial efforts in Nigeria, such as the amalgamation of the northern and southern protectorates in 1914. The results revealed significant discrepancies between Nigeria's northern and southern regions regarding cybercriminal arrest, conviction, and sentencing. Specifically, the results showed that the EFCC's criminalization of Southerners differs substantially from that of Northerners. The contemporary manifestation of inequalities concerning the production and prosecution of cybercrime on Twitter reflects long-standing contestations (e.g., economic, political, cultural, geological) between the northern and southern parts of Nigerian society. Therefore, since the North-South divide in present-day Nigeria originated from British colonization, colonialism is the base that shaped the superstructure comprising political, religious, historical, geological (e.g., crude oil), and economic factors. In turn, the article spotlights that regional differences in educational attainment, originating from differing experiences of Christianization and colonization, interact with regional disparities in the production of cybercrime. More research is required to better understand how these contextual disparities in society interact with the production and prosecution of cybercrime, given that Nigerian cybercriminals defraud victims all over the world.

Advantageous comparison: using Twitter responses to understand similarities between cybercriminals ("Yahoo Boys") and politicians ("Yahoo men").

This article is about the manifestations of similarities between two seemingly distinct groups of Nigerians: cybercriminals and politicians. Which linguistic strategies do Twitter users use to express their opinions on cybercriminals and politicians? The study undertakes a qualitative analysis of 'engaged' tweets of an elite law enforcement agency in West Africa. We analyzed and coded over 100,000 'engaged' tweets based on a component of mechanisms of moral disengagement (i.e., advantageous comparison), a linguistic device. The results reveal how respondents defend the actions of online fraudsters ("the deviant group") by strategically comparing them to the wrongful acts of corrupt politicians ("the respectable group"). Similarly, the results show how respondents positioned this linguistic strategy to compare "the powerless group" (online fraudsters) and "the powerful group" (politicians) in society. Indeed, tweet responses suggest that the Economic and Financial Crimes Commission (EFCC) generally looks downwards for culprits (i.e., online fraudsters) while ignoring fraudulent politicians. We conclude that the process by which some actions are interpreted as a crime compared to others is a moral enterprise.

Silver nanomaterials released from commercial textiles have minimal impacts on soil microbial communities at environmentally relevant concentrations.

Silver nanomaterials (Ag NMs) have been used in a variety of commercial products to take advantage of their antimicrobial properties. However, there are concerns that these AgNMs can be released during/after use and enter wastewater streams, potentially impacting aquatic systems or accumulating in wastewater biosolids. Biosolids, which are a residual of wastewater treatment processes, have been found to contain AgNMs and are frequently used as agricultural fertilizer. Since the function of soil microbial communities is imperative to nutrient cycling and agricultural productivity, it is important to characterize and assess the effects that silver nanomaterials could have in agricultural soils. In this study agricultural soil was amended with pristine engineered (PVP-coated or uncoated AgNMs), aged silver (sulphidized or released from textiles) nanomaterials, and ionic silver to determine the fate and toxicity over the course of three months. Exposures were carried out at various environmentally relevant concentrations (1 and 10 mg Ag/kg soil) representing between 30 to over 800 years of equivalent biosolid loadings. Over thirteen different methodologies and measures were used throughout this study to assess for potential effects of the silver nanomaterials on soil, including microbial community composition, average well colour development (AWCD) and enzymatic activity. Overall, the AgNM exposures did not exhibit significant toxic effects to the soil microbial communities in terms of density, activity, function and diversity. However, the positive ionic silver treatment (100 mg Ag/kg soil) resulted in suppression to microbial activity while also resulting in significantly higher populations of Frankia alni (nitrogen-fixer) and Arenimonas malthae (phytopathogen) as compared to the negative control (p < 0.05, Tukey HSD) which warrants further investigation.

DNA damage in earthworms from highly contaminated soils: assessing resistance to arsenic toxicity by use of the Comet assay.

Earthworms native to the former mine site of Devon Great Consols (DGC), UK reside in soils highly contaminated with arsenic (As). These earthworms are considered to have developed a resistance to As toxicity. The mechanisms underlying this resistance however, remain unclear. In the present study, non-resistant, commercially sourced Lumbricus terrestris were exposed to a typical DGC soil in laboratory mesocosms. The earthworms bio-accumulated As from the soil and incurred DNA-damage levels significantly above those observed in the control mesocosm (assessed using the Comet assay). A dose response was observed between DNA damage (% tail DNA) and As concentration in soil (control, 98, 183, 236, 324 and 436mgkg(-1)). As-resistant earthworms (Lumbricus rubellus, Dendrodrilus rubidus and L. terrestris) collected from contaminated soils at DGC (203 to 9025mgkg(-1) As) had also bio-accumulated high levels of As from their host soils, yet demonstrated low levels of DNA damage compared with earthworms from uncontaminated sites. The results demonstrate that the As-contaminated soils at DGC are genotoxic to non-native earthworms and much less so to earthworms native to DGC, thus providing further evidence of an acquired resistance to As toxicity in the native earthworms.

Assessing the role of microbial communities in the performance of constructed wetlands used to treat combined sewer overflows.

Combined sewer overflows are contaminated with various micropollutants which pose risk to both environmental and human health. Some micropollutants, such as carbamazepine and sulfamethoxazole, are very persistent and difficult to remove from wastewater. Event loaded vertical-flow constructed wetlands (retention soil filters; RSFs) have proven to be effective in the treatment of combined sewer overflows for a wide range of pollutants. However, little is known about how microbial communities contribute to the treatment efficiency, specifically to the reduction of micropollutants. To the best of our knowledge, this is the first study attempting to close this gap. Microbial communities in pilot-scale RSFs were investigated, which showed explicit grouping of metabolic activity at different filter depths with some differential abundance of identified genera. The highest microbial activity was found in the top layer of 0.75 m deep filters, whereas homogeneous activity dominated in a 0.50 m deep filter, indicating oxygen availability to be a limiting factor of the metabolic activity in RSFs. The removal efficiencies of all investigated organic trace substances were correlated to the utilization of specific carbon sources. Most notable is the correlation between the carbon source glucose-1-phosphate and the removal of metoprolol. The strongest correlations for other substances were the removal of diclofenac to the utilization of the carbohydrate i-erythritole; bisphenol A to carbohydrate α-d-lactose, and 1-H-benzotriazole to carbonic acid D-galacturonic acid. Those results are supported by positive correlations of specific microbial genera with both the utilization of the above mentioned carbon sources and the removal efficiency for the respective micropollutants. Most notable is correlation of Tetrasphaera and the removal of benzotriazole and diclofenac.

Human toenails as a biomarker of exposure to elevated environmental arsenic.

A pilot study was conducted to determine the applicability of toenails as a biomarker of exposure to elevated environmental arsenic (As) levels. A total of 17 individuals were recruited for the pilot study: 8 residents living near to a former As mine, Devon, UK, forming the exposed group, plus 9 residents from Nottinghamshire, UK, with no anticipated As exposure who were used for comparison as a control group. All toenail samples were thoroughly washed prior to analysis and the wash solutions retained for As determination via ICP-MS to provide an indication of the background environmental As levels for each group. Total As was determined in washed toenail samples via ICP-MS following microwave assisted acid digestion. Concentrations of total As in the toenails of the exposed group were elevated, ranging from 858 to 25 981 microg kg(-1) (geometric mean = 5406 microg kg(-1)), compared to the control group whose toenail As concentrations ranged from 73 to 273 microg kg(-1) (geometric mean = 122 microg kg(-1)). Higher levels of exogenous As contamination were present on the toenails of the exposed group (geometric mean = 506 microg kg(-1)) compared to the control group (geometric mean = 4.0 microg kg(-1)) providing evidence of higher environmental As levels in the exposed group. Total As concentrations in toenail samples were positively correlated to environmental As levels (r = 0.60, p < 0.001). HPLC-ICP-MS analysis of aqueous toenail extracts revealed inorganic arsenite (As(III)) to be the dominant species extracted ( approximately 83%) with lesser amounts of inorganic arsenate (As(V)) and organic dimethylarsinate (DMA(V)) at approximately 13% and approximately 8.5%, respectively. Arsenic speciation in analysed toenail extracts from the two groups was comparable. The only notable difference between groups was the presence of small amounts (<1%) of organic methylarsonate (MA(V)) in two toenail samples from the exposed group. Toenails are presented as a viable biomarker of exposure at sites with elevated environmental As, such as the former mining sites found throughout Devon and Cornwall, UK.

Arsenic biotransformation in earthworms from contaminated soils.

Two species of arsenic (As) resistant earthworm, Lumbricus rubellus and Dendrodrillus rubidus, their host soils and soil excretions (casts) were collected from 23 locations at a former As mine in Devon, UK. Total As concentrations, measured by ICP-MS, ranged from 255 to 13,080 mg kg(-1) in soils, 11 to 877 mg kg(-1) in earthworms and 284 to 4221 mg kg(-1) in earthworm casts from a sub-sample of 10 of the 23 investigated sites. The samples were also measured for As speciation using HPLC-ICP-MS to investigate potential As biotransformation pathways. Inorganic arsenate (As(V)) and arsenite (As(III)) were the only species detected in the soil. As(V) and As(III) were also the dominant species found in the earthworms and cast material together with lower proportions of the organic species methylarsonate (MA(V)), dimethylarsinate (DMA(V)), arsenobetaine (AB) and three arsenosugars. Whilst the inorganic As content of the earthworms increased with increasing As body burden, the concentration of organic species remained relatively constant. These results suggest that the biotransformation of inorganic arsenic to organic species does not contribute to As resistance in the sampled earthworm populations. Quantification of As speciation in the soil, earthworms and cast material allows a more comprehensive pathway for the formation of AB in earthworms to be elucidated.

Earthworms and in vitro physiologically-based extraction tests: complementary tools for a holistic approach towards understanding risk at arsenic-contaminated sites.

The relationship of the total arsenic content of a soil and its bioaccumulation by earthworms (Lumbricus rubellus and Dendrodrilus rubidus) to the arsenic fraction bioaccessible to humans, measured using an in vitro physiologically-based extraction test (PBET), was investigated. Soil and earthworm samples were collected at 24 sites at the former arsenic mine at the Devon Great Consols (DGC) in southwest England (UK), along with an uncontaminated site in Nottingham, UK, for comparison. Analysis of soil and earthworm total arsenic via inductively coupled plasma mass spectrometry (ICP-MS) was performed following a mixed acid digestion. Arsenic concentrations in the soil were elevated (204-9,025 mg kg(-1)) at DGC. The arsenic bioaccumulation factor (BAF) for both earthworm species was found to correlate positively with the human bioaccessible fraction (HBF), although the correlation was only significant (P < or = 0.05) for L. rubellus. The potential use of both in vitro PBETs and earthworms as complementary tools is explored as a holistic and multidisciplinary approach towards understanding risk at contaminated sites. Arsenic resistant earthworm species such as the L. rubellus populations at DGC are presented as a valuable tool for understanding risk at highly contaminated sites.

Impacts and fate of triclosan and sulfamethoxazole in intensified re-circulating vertical flow constructed wetlands.

The impacts to microbial function, overall performance and eventual fate were assessed for triclosan (TCL) and sulfamethoxazole (SMX) in intensified (re-circulating) vertical subsurface flow (VSSF) constructed wetlands (CWs). The potential toxicity of each pharmaceutical to the intrinsic microbial communities was first assessed over a wide exposure range (0-1000 µg/l) via an ex-situ dose-response assay to estimate the concentration at which adverse effects were likely to occur. Based on these results an acute (7 day) in-situ exposures (500 µg/l) were then performed and impacts to the mesocosm systems monitored for 1 month via community-level physiological profiling (CLPP) alongside chemical oxygen demand (COD) removal rates and a range of water quality, and hydrological parameters. Despite the clear potential for negative impacts to microbial function from both compounds observed at 100 µg/l in the ex-situ dose-response test, no impacts were observed for the 500 µg/l in-situ exposure in the VSSF mesocosms. COD removal, water chemistry, plant health, and hydrological parameters did not significantly change in response to the in-situ exposure. In terms of fate, the removal efficiency for both TCL and SMX was high (>80%) after 1 h and complete removal (>99.7%) was observed after 168 h. Following the in-situ exposure, and subsequent one month effects-monitoring period, the mesocosms were decommissioned with the media biofilm spatially assessed for organic content as well as TCL and SMX concentrations. TCL and SMX were found to have persisted in the media and demonstrated spatial variation with an overall 2-20% and 5-6% recovered respectively. This suggests that biofilm bound TCL and SMX were biologically degraded in VSSF CWs, however may also accumulate in the biofilm if TCL and SMX are maintained in the influent. These results reinforce the robustness and potential of constructed wetlands for the treatment of pharmaceutical and personal care product (PPCP) contaminated wastewater.

Impact of engineered nanoparticles on microbial transformations of carbon, nitrogen, and phosphorus in wastewater treatment processes - A review.

Concern regarding the potential negative impacts of released engineered nanoparticles (ENPs) on pollutant removal performance of wastewater treatment systems has received booming attention in recent years. However, the conclusions drawn from different studies often lead to fragmented overall knowledge, some of which are even contradictory. This scenario shows the necessity for a comprehensive review of the interactions of ENPs in wastewater treatment systems, particularly on the impacts of ENPs on microbial processes of carbon (C), nitrogen (N), and phosphorus (P) removal in water treatment systems. This review introduced the impact of 6 often reported ENPs in 5 types of treatment systems. We found that exposure to most of the investigated ENPs at low concentrations doesn't adversely influence the growth of the heterotrophic microbes, which are responsible for organic matter removal. The impacts of ENPs on various microbial nitrogen transformation processes have been investigated. Dosing of ENPs often causes acute microbial nitrogen removal inhibition at various concentrations, but does not influence long-term operation due to microbial adaption. No significant negative effects on biological phosphorus removal in different wastewater treatment processes have been reported after both short-term and long-term exposure (except copper nanoparticles). Environmentally relevant concentrations of ENPs have been reported to enhance the photosynthetic capacity of wetland plants, whereas chronic inhibition to photosynthesis was found in exposure to high concentrations of ENPs. Inhibition effects are often overestimated in pure cultivated toxicity test assays compared to testing with artificially prepared wastewater containing various ingredients or with real wastewater. Potential ligands in real wastewater can bind with ENPs and lower their dissolution. Some challenges exist regarding detection and quantification techniques of ENPs at environmental concentrations, modeling of engineered nanomaterial release on a worldwide scale, and inhibitory mechanisms to microbial transformations.

Interstitial water microbial communities as an indicator of microbial denitrifying capacity in wood-chip bioreactors.

The discharge from food production greenhouses (greenhouse effluent) contains high nutrient and salt concentrations, which, if directly released, can have adverse effects on the environment. Wood-chip bioreactors are increasingly popular passive water treatment systems favoured for their economical denitrification in treating agricultural field tile drainage. Microbial communities are central to denitrification; however little is known about the maturation of microbial communities in wood-chip bioreactors treating greenhouse effluents. In this study, multiple subsurface flow wood-chip bioreactors, each vegetated with a different plant species, together with an unplanted unit, received synthetic greenhouse effluent with elevated nitrate concentrations. The hybrid bioreactors were operated for over 2 years, during which time water samples were collected from the inlet, outlet and within the reactors. The increasing denitrification rate in the bioreactor planted with Typha angustifolia (narrowleaf cattail) correlated with increasing microbial activity and metabolic richness, measured by the carbon utilization patterns in Biolog® EcoPlates. Increased denitrifying gene (nirS) copies (determined by quantitative polymerase chain reaction, qPCR), and near-complete nitrate removal were observed in the T. angustifolia and unplanted reactors after 16 and 23 months of operation respectively. The findings suggested that an acclimation period of at least one year can be expected in unseeded bioreactors planted with T. angustifolia, while bioreactors without vegetation may require a longer time to maximize their denitrification capacity. These results are important for the design and operation of wood-chip bioreactors, which are expected to be more commonly applied in the future.

Microbial community metabolic profiles in saturated constructed wetlands treating iohexol and ibuprofen.

The aim of the present study was to elucidate the microbial community metabolic profiles in saturated constructed wetland (CW) mesocosms planted with five different wetland plant species fed with water individually spiked with 100 µg L-1 ibuprofen or iohexol. Community-level physiological profiling (CLPP) using Biolog Ecoplates was performed and coupled with the assessment of water quality parameters (water temperature, pH, DO and TOC, TN, NH4-N, PO4-P removal efficiency). The microbial community metabolic profiles (microbial activity, richness, and carbon source utilization), as well as the water quality parameters revealed similar trends among the control mesocosms and the mesocosms fed with water spiked with iohexol and ibuprofen. Significant differences were observed between the planted and unplanted mesocosms and between seasons (summer and winter) within each of the feeding lines (control, iohexol or ibuprofen). The microbial community metabolic profiles in the saturated CW were shaped by plant presence and plant species, while no negative impact of iohexol and ibuprofen presence was noticed at the 100 µg L-1. In addition, the microbial activity and richness were generally higher in planted mesocosms than in the unplanted systems in the summer. For the first time, a positive correlation between iohexol removal and the microbial community metabolic profiles (activity, richness and amines and amides utilization in summer, and carbohydrates utilization in winter) in the saturated mesocosms was observed. Putrescine utilization in the summer and d-cellobiose, d,l-alpha-glycerol phosphate in winter were linked with the metabolic processing of iohexol, while glycogen in summer and l-phenylalanine, Glycyl-l-glutamic acid in winter were linked with ibuprofen removal efficiency in the saturated CW.

Quantitative arsenic speciation in two species of earthworms from a former mine site.

The relationship between the total arsenic concentration and the chemical speciation of arsenic in two species of earthworm (Lumbricus rubellus and Dendrodrilus rubidus) in relation to the host soil, was investigated for 13 sites of varying arsenic content, including a background level garden soil and a former mine site at the Devon Great Consols, UK. Earthworms were collected with the host soil (As soil concentration range 16-12, 466 mg kg(-1) dry weight) and measured for their total arsenic (concentration range 7-595 mg kg(-1) dry weight) using inductively coupled plasma mass spectrometry (ICP-MS). A methanol-water mixture was used to extract arsenic species from the earthworms prior to determination of the individual arsenic species by a combination of anion and cation exchange high performance liquid chromatography coupled to inductively coupled plasma mass spectrometry (HPLC-ICP-MS). A gradient elution anion exchange method is presented whereby nine arsenic species could be measured in one sample injection. Arsenic species were identified by comparison of retention times and sample spiking with known standards and a fully characterised seaweed extract. Arsenic was generally present in the earthworm as arsenate (As(V)) or arsenite (As(III)) and arsenobetaine (AB). Methylarsonate (MA), dimethylarsinate (DMA) and three arsenosugars (glycerol, phosphate, sulfate) were present as minor constituents. These results are discussed in relation to the mechanisms for coping with exposure to soil bound arsenic.

Impacts of design configuration and plants on the functionality of the microbial community of mesocosm-scale constructed wetlands treating ibuprofen.

Microbial degradation is an important pathway during the removal of pharmaceuticals in constructed wetlands (CWs). However, the effects of CW design, plant presence, and different plant species on the microbial community in CWs have not been fully explored. This study aims to investigate the microbial community metabolic function of different types of CWs used to treat ibuprofen via community-level physiological profiling (CLPP) analysis. We studied the interactions between three CW designs (unsaturated, saturated and aerated) and six types of mesocosms (one unplanted and five planted, with Juncus, Typha, Berula, Phragmites and Iris) treating synthetic wastewater. Results show that the microbial activity and metabolic richness found in the interstitial water and biofilm of the unsaturated designs were lower than those of the saturated and aerated designs. Compared to other CW designs, the aerated mesocosms had the highest microbial activity and metabolic richness in the interstitial water, but similar levels of biofilm microbial activity and metabolic richness to the saturated mesocosms. In all three designs, biofilm microbial metabolic richness was significantly higher (p < .05) than that of interstitial water. Both the interstitial water and biofilm microbial community metabolic function were influenced by CW design, plant presence and species, but design had a greater influence than plants. Moreover, canonical correlation analysis indicated that biofilm microbial communities in the three designs played a key role in ibuprofen degradation. The important factors identified as influencing ibuprofen removal were microbial AWCD (average well color development), microbial metabolic richness, and the utilization of amino acids and amine/amides. The enzymes associated with co-metabolism of l-arginine, l-phenyloalanine and putrescine may be linked to ibuprofen transformations. These results provide useful information for optimizing the operational parameters of CWs to improve ibuprofen removal.

Functionality of microbial communities in constructed wetlands used for pesticide remediation: Influence of system design and sampling strategy.

The objective of this study was to compare the microbial community metabolic function from both unsaturated and saturated constructed wetland mesocosms (CWs) when treating the pesticide tebuconazole. The comparison was performed for both interstitial water and substrate biofilm by community level physiological profiling (CLPP) via BIOLOG™ EcoPlates. For each CW design (saturated or unsaturated), six mesocosms were established including one unplanted and five planted individually with either Juncus effusus, Typha latifolia, Berula erecta, Phragmites australis or Iris pseudacorus. Microbial activity and metabolic richness of interstitial water from unsaturated CWs were significantly lower than that from saturated CWs. However, in general, the opposite result was observed for biofilm samples. Wetland plants promoted significantly higher biofilm microbial activity and metabolic richness than unplanted CWs in both CW designs. Differences in the microbial community functional profiles between plant species were only found for saturated CWs. Biofilm microbial metabolic richness was generally statistically higher than that of interstitial water in both unsaturated (1.4-24 times higher) and saturated (1.2-1.7 times higher) CWs. Carbon source (guild) utilization patterns were generally different between interstitial water and biofilm samples. Functionality of the biofilm microbial community was positively correlated to the removal of all pollutants (TN, NH4+-N, TP, TOC and tebuconazole) for both unsaturated and saturated CWs, suggesting the biofilm plays a more important role in pollutant removal than the interstitial water microbial community. Thus, merely observing the interstitial water microbial communities may underestimate the role of the microbial community in CW performance. Interestingly, the ability for the biofilm microbial community to utilize amino acids and amines/amides was positively correlated with tebuconazole removal in all system types.

Community-Level Physiological Profiling of Microbial Communities in Constructed Wetlands: Effects of Sample Preparation.

Community-level physiological profiling (CLPP) using BIOLOG® EcoPlates™ has become a popular method for characterizing and comparing the functional diversity, functional potential, and metabolic activity of heterotrophic microbial communities. The method was originally developed for profiling soil communities; however, its usage has expanded into the fields of ecotoxicology, agronomy, and the monitoring and profiling of microbial communities in various wastewater treatment systems, including constructed wetlands for water pollution control. When performing CLPP on aqueous samples from constructed wetlands, a wide variety of sample characteristics can be encountered and challenges may arise due to excessive solids, color, or turbidity. The aim of this study was to investigate the impacts of different sample preparation methods on CLPP performed on a variety of aqueous samples covering a broad range of physical and chemical characteristics. The results show that using filter paper, centrifugation, or settling helped clarify samples for subsequent CLPP analysis, however did not do so as effectively as dilution for the darkest samples. Dilution was able to provide suitable clarity for the darkest samples; however, 100-fold dilution significantly affected the carbon source utilization patterns (CSUPs), particularly with samples that were already partially or fully clear. Ten-fold dilution also had some effect on the CSUPs of samples which were originally clear; however, the effect was minimal. Based on these findings, for this specific set of samples, a 10-fold dilution provided a good balance between ease of use, sufficient clarity (for dark samples), and limited effect on CSUPs. The process and findings outlined here can hopefully serve future studies looking to utilize CLPP for functional analysis of microbial communities and also assist in comparing data from studies where different sample preparation methods were utilized.

Transcriptomic Responses During Early Development Following Arsenic Exposure in Western Clawed Frogs, Silurana tropicalis.

Arsenic compounds are widespread environmental contaminants and exposure elicits serious health issues, including early developmental anomalies. Depending on the oxidation state, the intermediates of arsenic metabolism interfere with a range of subcellular events, but the fundamental molecular events that lead to speciation-dependent arsenic toxicity are not fully elucidated. This study therefore assesses the impact of arsenic exposure on early development by measuring speciation and gene expression profiles in the developing Western clawed frog (Silurana tropicalis) larvae following the environmental relevant 0.5 and 1 ppm arsenate exposure. Using HPLC-ICP-MS, arsenate, dimethylarsenic acid, arsenobetaine, arsenocholine, and tetramethylarsonium ion were detected. Microarray and pathway analyses were utilized to characterize the comprehensive transcriptomic responses to arsenic exposure. Clustering analysis of expression data showed distinct gene expression patterns in arsenate treated groups when compared with the control. Pathway enrichment revealed common biological themes enriched in both treatments, including cell signal transduction, cell survival, and developmental pathways. Moreover, the 0.5 ppm exposure led to the enrichment of pathways and biological processes involved in arsenic intake or efflux, as well as histone remodeling. These compensatory responses are hypothesized to be responsible for maintaining an in-body arsenic level comparable to control animals. With no appreciable changes observed in malformation and mortality between control and exposed larvae, this is the first study to suggest that the underlying transcriptomic regulations related to signal transduction, cell survival, developmental pathways, and histone remodeling may contribute to maintaining ongoing development while coping with the potential arsenic toxicity in S. tropicalis during early development.

Arsenic(+3) and DNA methyltransferases, and arsenic speciation in tadpole and frog life stages of western clawed frogs (Silurana tropicalis) exposed to arsenate.

Western clawed frog (Silurana tropicalis) embryos were exposed to control, low (nominally 0.5 mg L(-1)) and high (nominally 1 mg L(-1)) arsenate (As(V)) culture water concentrations to investigate the effects of arsenic (As) on different life stages, namely tadpole (Nieuwkoop and Faber stage 56, NF56) and frog stages (NF66). The effects were assessed by measuring arsenic(+3) and DNA methyltransferases (AS3MT and DNMT1), as well as As speciation in the tissues. The As content in frog tissues increased with water As concentration. The As species observed by high performance liquid chromatography - inductively coupled plasma mass spectrometry (HPLC-ICPMS) were mostly inorganic, dimethylarsinic acid (DMA) and trimethylarsine oxide (TMAO). With solid state X-ray absorption near edge structure (XANES) analysis, arsenobetaine/tetramethylarsonium ion were also seen. AS3MT levels decreased upon low As exposure in NF56, rising again to control levels at the high As exposure. In NF66 tissues, on the other hand, AS3MT decreased only with NF66 high As exposure. DNMT1 increased with exposure, and this was statistically significant only for the high As exposure at both life stages. Thus these enzymes seem to be affected by the As exposure. Methylation of As to form monomethylarsonate (MMA), DMA and TMAO in the frogs appeared to be inversely related to AS3MT levels. A possible interpretation of this finding is that when AS3MT is higher, excretion of MMA + DMA + TMAO is more efficient, leaving lower concentrations in the tissues, with the opposite effect (less excretion) when AS3MT is lower; alternatively, other enzymes or linked genes may affect the methylation of As.