Aquatic Phytotoxicity to Lemna minor of Three Commonly Used Drugs of Addiction in Australia.

The manufacturing and consumption of drugs of addiction has increased globally and their widespread occurrence in the environment is an emerging concern. This study evaluated the phytotoxicity of three compounds: methamphetamine, codeine and morphine; commonly reported in Australian urban water, to the aquatic plant Lemna minor under controlled conditions. L. minor was sensitive to lower drug concentrations when administered in multi-compound mixtures (100-500 µg L-1) than when applied individually (range 600-2500 µg L-1), while no adverse effects were observed at environmentally-relevant concentrations (1-5 µg L-1) detected in wastewater effluent. In conclusion, the results show that the concentrations of these compounds discharged into the environment are unlikely to pose adverse phytotoxic effects. These three compounds are known to be the most stable of their group under such conditions indicating that with this respect it is safe to use recycled water for existing regulated reclaimed purposes including agricultural or parklands irrigation or replenishing surface and groundwater. However, more research on the analysis of methamphetamines and opiates in municipal effluents is needed to reassure the likely environmental hazard of these neuroactive drug classes to aquatic organisms. Given the ever-growing production and aquatic disposal of discharge wastewater globally, this study provides timely and valuable insights into the likely drug-related impacts of effluent disposal on aquatic plants in receiving environments.

Sphingopyxis Species Isolated from Sand Filter Biofilm at an Australian Drinking Water Treatment Works.

Three strains isolated by geosmin enrichment from a sand filter in an Australian drinking water treatment works were genome sequenced to identify their taxonomic placement, and a bench-scale batch experiment confirmed their geosmin-degrading capability. Using the average nucleotide identity based on the MUMmer algorithm (ANIm), pairwise digital DNA-DNA hybridization (dDDH), and phylogenomic analyses, the strains were identified as Sphingopyxis species.

Removal of cyanobacterial metabolites through wastewater treatment plant filters.

Wastewaters have the potential to proliferate excessive numbers of cyanobacteria due to high nutrient levels. This could translate to the production of metabolites, such as the saxitoxins, geosmin and 2-methylisoborneol (MIB), which can impair the quality of wastewater destined for re-use. Biological sand filtration was assessed for its ability to remove these metabolites from a wastewater. Results indicated that the sand filter was incapable of effectively removing the saxitoxins and in some instances, the effluent of the sand filter displayed greater toxicity than the influent. Conversely, the sand filter was able to effectively remove geosmin and MIB, with removal attributed to biodegradation. Granular activated carbon was employed as an alternative filter medium to remove the saxitoxins. Results showed similar removals to previous drinking water studies, where efficient removals were initially observed, followed by a decrease in the removal; a consequence of the presence of competing organics which reduced adsorption of the saxitoxins.

Stormwater monitoring using on-line UV-Vis spectroscopy.

Stormwater runoff contains a myriad of pollutants, including faecal microbes, and can pose a threat to urban water supplies, impacting both economic development and public health. Therefore, it is a necessity to implement a real-time hazard detection system that can collect a substantial amount of data, assisting water authorities to develop preventive strategies to ensure the control of hazards entering drinking water sources. An on-line UV-Vis spectrophotometer was applied in the field to collect real-time continuous data for various water quality parameters (nitrate, DOC, turbidity and total suspended solids) during three storm events in Mannum, Adelaide, Australia. This study demonstrated that the trends for on-line and comparative laboratory-analysed samples were complimentary through the events. Nitrate and DOC showed a negative correlation with water level, while turbidity and total suspended solids indicated a positive correlation with water level during the high rainfall intensity. The correlations among nitrate, DOC, turbidity, total suspended solids and water level are the opposite during low rainfall intensity. Nitrate, one of the main pollutants in stormwater, was investigated and used as a surrogate parameter for microbial detection. However, the microbiological data (Escherichia coli) from captured storm events showed poor correlations to nitrate and other typical on-line parameters in this study. This is possibly explained by the nature of the stormwater catchment outside of rain events, where the sources of bacteria and nutrients may be physically separated until mixed during surface runoff as a result of rainfall. In addition, the poor correlations among the microbiological data and on-line parameters could be due to the different sources of bacteria and nutrients that were transported to the stormwater drain where sampling and measurement were conducted.

Development of an mlrA gene-directed TaqMan PCR assay for quantitative assessment of microcystin-degrading bacteria within water treatment plant sand filter biofilms.

We report for the first time a quantitative mlrA gene-directed TaqMan PCR assay for the rapid detection of microcystin-degrading bacteria. This was applied, in combination with 16S ribosomal DNA-directed quantitative PCR and denaturing gradient gel electrophoresis, to study virgin sand filter column biofilm development and to correlate mlrA gene abundance with microcystin removal efficiency.

Enhancing the biofiltration of geosmin by seeding sand filter columns with a consortium of geosmin-degrading bacteria.

Geosmin is a secondary metabolite that can be produced by many species of cyanobacteria and Actinomycetes. It imparts a musty/earthy taste and odour to drinking water which can result in consumer complaints and a general perception that there is a problem with the water quality. As geosmin is recalcitrant to conventional water treatment, processes are sought to ensure effective removal of this compound from potable water. Biological filtration (biofiltration) is an attractive option for geosmin removal as this compound has been shown to be biodegradable. However, effective biofiltration of geosmin can be site specific as it is highly dependent upon the types of organism present and there is often an extended acclimation period before efficient removals are achieved. We report here, a novel approach to enhance the biofiltration of geosmin by seeding sand filter columns with a bacterial consortium previously shown to be capable of effectively degrading geosmin. Geosmin removals of up to 75% were evident through sand columns which had been inoculated with the geosmin-degrading bacteria, when compared with non-inoculated sand columns where geosmin removals were as low as 25%. These low geosmin removals through the non-inoculated sand columns are consistent with previous studies and were attributed to physical/abiotic losses. The presence of an existing biofilm was shown to influence geosmin removal, as the biofilm allowed for greater attachment of the geosmin-degrading consortium (as determined by an ATP assay), and enhanced removals of geosmin. Minimal difference in geosmin removal was observed when the geosmin-degrading bacteria were inoculated into the sand columns containing either an active or inactive biofilm.

Biodegradation of geosmin by a novel Gram-negative bacterium; isolation, phylogenetic characterisation and degradation rate determination.

Biologically active sand filters within water treatment plants (WTPs) are now recognised as an effective barrier for the removal of geosmin. However, little is known regarding the actual microbiological processes occurring or the bacteria capable of degrading geosmin. This study reports the enrichment and isolation of a Gram-negative bacterium, Geo48, from the biofilm of a WTP sand filter where the isolate was shown to effectively degrade geosmin individually. Experiments revealed that Geo48 degraded geosmin in a planktonic state by a pseudo-first-order mechanism. Initial geosmin concentrations ranging from 100 to 1000ng/l were shown to directly influence geosmin degradation in reservoir water by Geo48, with rate constants increasing from 0.010h(-1) (R(2)=0.93) to 0.029h(-1) (R(2)=0.97) respectively. Water temperature also influenced degradation of geosmin by Geo48 where temperatures of 11, 22 and 30 degrees C resulted in rate constants of 0.017h(-1) (R(2)=0.98), 0.023h(-1) (R(2)=0.91) and 0.019h(-1) (R(2)=0.85) respectively. Phylogenetic analysis using the 16S rRNA gene of Geo48 revealed it was a member of the Alphaproteobacteria and clustered with 99% bootstrap support with an isolate designated Geo24, a Sphingopyxis sp. previously described as degrading geosmin but only as a member of a bacterial consortium. Of the previously described bacteria, Geo48 was most similar to Sphingopyxis alaskensis (97.2% sequence similarity to a 1454bp fragment of the 16S rRNA gene). To date, this is the only study to report the isolation and characterisation of a Gram-negative bacterium from a biologically active sand filter capable of the sole degradation of geosmin.

Label-Free Bacterial Toxin Detection in Water Supplies Using Porous Silicon Nanochannel Sensors.

Lipopolysaccharides (LPS) are the major component of the outer membrane of all Gram-negative bacteria and some cyanobacteria and are released during growth and cell death. LPS pose a potential health risk in water, causing acute respiratory illnesses, inhalation fever, and gastrointestinal disorders. The need for rapid and accurate detection of LPS has become a major priority to facilitate more timely and efficacious intervention and, hence, avoid unsafe water distribution. In this context, a porous silicon membrane (pSiM)-based electrochemical biosensor was developed for direct and sensitive detection of LPS. pSiM, featuring arrays of nanochannels, was modified with polymyxin B (PmB), an antimicrobial peptide with strong affinity to LPS. Detection of LPS was based on measuring the changes in the diffusion through the nanochannels of an electroactive species added in solution, caused by the nanochannel blockage upon LPS binding to PmB. Results showed a limit of detection of 1.8 ng/mL, and a linear response up to 10,000 ng/mL spiked in buffer. Selectivity of the sensor toward potential interfering species in water supplies was also assessed. Sensor performance was then evaluated in water samples from a water treatment plant (WTP), and detection of LPS well below the levels encountered in episodes of water contamination and in humidifiers was demonstrated. The same platform was also tested for bacterial detection including Pseudomonas aeruginosa and Escherichia coli spiked in water samples from a WTP. Considering its performance characteristics, this platform represents a promising screening tool to identify the presence of LPS in water supplies and provide early warning of contamination events.

Transformation pathway and toxicity assessment of malathion in aqueous solution during UV photolysis and photocatalysis.

In drinking water treatment, complete mineralization of organophosphorus pesticides (OPPs) by UV-based advanced oxidation processes (UV AOPs) is rarely achieved. The formation of intermediate oxidation byproducts would likely have some profound effects on toxicity of the reaction solutions. This study investigated the intermediate oxidation byproducts, transformation pathway and toxicity of malathion solutions during the treatment processes of UV alone, UV/H2O2, UV/TiO2 and UV/Fenton. The main intermediate oxidation byproducts were derived using ultra-performance liquid chromatography - electrospray - time-of-flight mass spectrometry. Thereby the transformation pathway for each of these treatment processes was proposed. The results indicate that in UV photolysis, the transformation pathway of malathion proceeded initially via cleavage of the phosphorus-sulfur bonds while in photocatalysis, the desulfurization from a PS bond to a PO bond was the primary degradation pathway. Interestingly, only in the UV/TiO2 process a small fraction of malathion was found decomposed via a demethylation reaction. At the same time, a toxicity assessment of the treated solutions was conducted by both luminescence inhibition of Vibrio fischeri and inhibition of acetylcholinesterase (AChE). It was found that after UV AOP treatment, the toxicity of the malathion aqueous solution increased sharply. In contrast, no increase in toxicity was observed for the malathion aqueous solution after UV alone treatment. This study demonstrates that the high removal efficiency achieved by OPPs does not imply that detoxification of the water solution has been achieved. On the contrary, the toxicity of the treated solutions by OPPs may be increased significantly depending on the selected treatment processes.

Removal of emerging drugs of addiction by wastewater treatment and water recycling processes and impacts on effluent-associated environmental risk.

Drugs of addiction, have been recognized as potential contaminants of concern to the environment. Effluent wastewater discharge is a major source of contamination to aquatic receiving environments. A year-long monitoring program was undertaken in Australia to characterise the fate of four emerging drugs of addiction: methamphetamine; MDMA; pharmaceutical opioids: codeine and morphine and a metabolite: benzoylecgonine in four wastewater treatment plants operating with different secondary treatment technologies: conventional activated sludge (CAS), membrane bioreactors (MBR), integrated fixed-film AS (IFAS) and sequencing batch reactor (SBR). The effect of subsequent tertiary treatment (coagulation/flocculation) on the removal efficiency was also assessed. Drugs were detected in influent and effluent samples (mean concentration ranged from 43-4777 and 17-1721 ng/L, respectively). Treated effluents had noticeably lower levels compared to raw influents. Removal efficiency of compounds depended on the secondary treatment employed, with IFAS and MBR performing the best with significant removal of compounds (≈90%) followed by CAS (54-96%) and lastly SBR (42-83%). Despite the low levels of drugs measured after the secondary treatment, near complete removal after tertiary treatment (≈99%) was recorded, which demonstrated the effectiveness of using the coagulation/flocculation process as an effective step for enhancing the removal efficiency. The levels of drugs were at a low level in the effluents released into the environment and used for recycling and all posed a low environmental risk in urban water courses based on the risk assessment. The information given here provides new and useful information to the water industry and regulators on the efficiency of drug removal in a range of wastewater treatment configurations.

Occurrence, removal and environmental risk of markers of five drugs of abuse in urban wastewater systems in South Australia.

The occurrence and fate of five drugs of abuse in raw influent and treated effluent wastewater were investigated over a period of 1 year in the Adelaide region of South Australia. Four wastewater treatment plants were chosen for this study and monitored for five drugs which included cocaine in the form of its metabolite benzoylecgonine (BE), methamphetamine, 3,4-methylenedioxymethamphetamine (MDMA) and two opioids (codeine and morphine) during the period April 2016 to February 2017. Alongside concentrations in raw sewage, the levels of drugs in the treated effluent were assessed and removal efficiencies were calculated. Drug concentrations were measured by mixed-mode solid phase extraction and liquid chromatography coupled to a quadrupole mass spectrometer. Drug concentrations detected in the raw wastewater ranged from 7 to 6510 ng/L and < LOD to 4264 ng/L in treated effluent samples. Drug removal rates varied seasonally and spatially. The mass loads of drugs discharged into the environment were in descending order: codeine > methamphetamine > morphine > MDMA > BE. Results showed that all the targeted drugs were on average incompletely removed by wastewater treatment, with removal performance highest for morphine (94%) and lowest for MDMA (58%). A screening-level environmental risk assessment was subsequently performed for the drugs based on effluent wastewater concentrations. Based on calculated risk quotients, overall environmental risk for these compounds appears low, with codeine and methamphetamine likely to pose the greatest potential risk to receiving environments. Given the recognised limitations of current ecotoxicological models and risk assessment methods for these and other pharmaceutical drugs, the potential for environmental impacts associated with the continuous discharge of these compounds in wastewater effluents should not be overlooked.

Effect of water treatment processes on Cryptosporidium infectivity.

Conventional water treatment processes have the ability to remove Cryptosporidium oocysts through coagulation, flocculation, sedimentation and filtration, provided there is efficient management of plant performance. The potential exists for the breakthrough of oocysts through the treatment train. The effect of the water treatment chemical aluminium sulphate (alum) on Cryptosporidium oocyst infectivity has been assessed using an assay that combines cell culture and real-time polymerase chain reaction techniques. The infectivity of fresh and temperature-aged oocysts (stored up to 6 months at 4 or 15 degrees C) was unaffected by exposure to a range of doses of alum in standard jar test procedures and dissolved air flotation processes and subsequent exposure to chlorine or chloramine. Removal efficiencies and infectivity measures are important in determining risk to public health and will reflect the ability of water treatment plants to act as a barrier to these pathogens.

Removal of glyphosate and aminomethylphosphonic acid from synthetic water by nanofiltration.

The removal of glyphosate and aminomethylphosphonic acid (AMPA) with synthetic water was carried out on a lab-scale nanofiltration unit using two membranes, NFX and NFW. The presence of humic acid and some inorganic matters (CaCl2 and NaCl) was tested in the experiment. The results demonstrate that NFX exhibits better separation performance than NFW. The herbicide filtration is found to have little effect on the permeate flux as compared to transmembrane pressure. Intermediate concentrations of NaCl act positively on foulant separation, and an increment of 3.3-5.4 percentage points in foulant rejection is obtained with the addition of 100 mg/L of NaCl. In Contrast, CaCl2 has negative effect on foulant separation during nanofiltration. Humic acid alone shows little influence on the rejection performance, whereas it is slightly improved in the coexistence of humic acid and CaCl2. The nanofiltration technology proves to be a good approach to treat the problem of pesticide micropollution in a one-step process. This work clearly shows that the composition of the water matrices may influence the efficiency of the nanofiltration process in terms of the separation of the micropollutants.

Nanostructured Electrochemical Biosensors for Label-Free Detection of Water- and Food-Borne Pathogens.

The emergence of nanostructured materials has opened new horizons in the development of next generation biosensors. Being able to control the design of the electrode interface at the nanoscale combined with the intrinsic characteristics of the nanomaterials engenders novel biosensing platforms with improved capabilities. The purpose of this review is to provide a comprehensive and critical overview of the latest trends in emerging nanostructured electrochemical biosensors. A detailed description and discussion of recent approaches to construct label-free electrochemical nanostructured electrodes is given with special focus on pathogen detection for environmental monitoring and food safety. This includes the use of nanoscale materials such as nanotubes, nanowires, nanoparticles, and nanosheets as well as porous nanostructured materials including nanoporous anodic alumina, mesoporous silica, porous silicon, and polystyrene nanochannels. These platforms may pave the way toward the development of point-of-care portable electronic devices for applications ranging from environmental analysis to biomedical diagnostics.

Use of DNA melting simulation software for in silico diagnostic assay design: targeting regions with complex melting curves and confirmation by real-time PCR using intercalating dyes.

BACKGROUND: DNA melting curve analysis using double-stranded DNA-specific dyes such as SYTO9 produce complex and reproducible melting profiles, resulting in the detection of multiple melting peaks from a single amplicon and allowing the discrimination of different species. We compare the melting curves of several Naegleria and Cryptosporidium amplicons generated in vitro with in silico DNA melting simulations using the programs POLAND and MELTSIM., then test the utility of these programs for assay design using a genetic marker for toxin production in cyanobacteria. RESULTS: The SYTO9 melting curve profiles of three species of Naegleria and two species of Cryptosporidium were similar to POLAND and MELTSIM melting simulations, excepting some differences in the relative peak heights and the absolute melting temperatures of these peaks. MELTSIM and POLAND were used to screen sequences from a putative toxin gene in two different species of cyanobacteria and identify regions exhibiting diagnostic melting profiles. For one of these diagnostic regions the POLAND and MELTSIM melting simulations were observed to be different, with POLAND more accurately predicting the melting curve generated in vitro. Upon further investigation of this region with MELTSIM, inconsistencies between the melting simulation for forward and reverse complement sequences were observed. The assay was used to accurately type twenty seven cyanobacterial DNA extracts in vitro. CONCLUSION: Whilst neither POLAND nor MELTSIM simulation programs were capable of exactly predicting DNA dissociation in the presence of an intercalating dye, the programs were successfully used as tools to identify regions where melting curve differences could be exploited for diagnostic melting curve assay design. Refinements in the simulation parameters would be required to account for the effect of the intercalating dye and salt concentrations used in real-time PCR. The agreement between the melting curve simulations for different species of Naegleria and Cryptosporidium and the complex melting profiles generated in vitro using SYTO9 verified that the complex melting profile of PCR amplicons was solely the result of DNA dissociation. Other data outputs from these simulations were also used to identify the melting domains that contributed to the observed melting peaks for each of the different PCR amplicons.

Isolation and identification of a novel microcystin-degrading bacterium from a biological sand filter.

A novel bacterium capable of degrading two microcystin analogues, microcystin-LR and -LA (MCLR and MCLA), was isolated from a biological sand filter which was previously shown to effectively remove these toxins from source waters. Based on phylogenetic analysis of the 16S rRNA gene sequence, the isolated organism, LH21, most likely belonged to the genus Sphingopyxis and of the previously cultured species clustered with Sphingopyxis witflariensis. Using polymerase chain reaction (PCR), isolate LH21 was shown to contain homologues to each of the four genes, mlrA, mlrB, mlrC and mlrD previously associated with the degradation of MCLR by Sphingomonas sp. ACM-3962. Isolate LH21 was able to effectively degrade MCLR and MCLA in batch experiments under environmentally relevant conditions, with complete removal observed within 5h after re-exposure of the toxins.

Biodegradation rates of 2-methylisoborneol (MIB) and geosmin through sand filters and in bioreactors.

Taste and odour (T&O) causing compounds, in particular, 2-methylisoborneol (MIB) and geosmin, are a problem for water authorities as they are recalcitrant to conventional water treatment. In this study, biological sand filtration was shown to be an effective process for the complete removal of MIB and geosmin, with removal shown to be predominantly through biodegradation. In addition, MIB and geosmin were also effectively degraded in batch bioreactor experiments using biofilm sourced from one of the sand filters as the microbial inoculum. The biodegradation of MIB and geosmin was determined to be a pseudo-first-order reaction with rate constants ranging between 0.10 and 0.58 d(-1) in the bioreactor experiments. Rate constants were shown to be dependent upon the initial concentration of the microbial inoculum but not the initial concentration of MIB and geosmin when target concentrations of 200 and 50 ng l(-1) were used. Furthermore, rate constants were shown to increase upon re-exposure of the biofilm to both T&O compounds. Enrichment cultures with subsequent community profile analysis using 16S rRNA-directed PCR-DGGE identified four bacteria most likely involved in the biodegradation of geosmin within the sand filters and bioreactors. These included a Pseudomonas sp., Alphaproteobacterium, Sphingomonas sp. and an Acidobacteriaceae member.

Occurrence of illicit drugs in water and wastewater and their removal during wastewater treatment.

This review critically evaluates the types and concentrations of key illicit drugs (cocaine, amphetamines, cannabinoids, opioids and their metabolites) found in wastewater, surface water and drinking water sources worldwide and what is known on the effectiveness of wastewater treatment in removing such compounds. It is also important to amass information on the trends in specific drug use as well as the sources of such compounds that enter the environment and we review current international knowledge on this. There are regional differences in the types and quantities of illicit drug consumption and this is reflected in the quantities detected in water. Generally, the levels of illicit drugs in wastewater effluents are lower than in raw influent, indicating that the majority of compounds can be at least partially removed by conventional treatment processes such as activated sludge or trickling filters. However, the literature also indicates that it is too simplistic to assume non-detection equates to drug removal and/or mitigation of associated risks, as there is evidence that some compounds may avoid detection via inadequate sampling and/or analysis protocols, or through conversion to transformation products. Partitioning of drugs from the water to the solids fraction (sludge/biosolids) may also simply shift the potential risk burden to a different environmental compartment and the review found no information on drug stability and persistence in biosolids. Generally speaking, activated sludge-type processes appear to offer better removal efficacy across a range of substances, but the lack of detail in many studies makes it difficult to comment on the most effective process configurations and operations. There is also a paucity of information on the removal effectiveness of alternative treatment processes. Research is also required on natural removal processes in both water and sediments that may over time facilitate further removal of these compounds in receiving environments.

A Novel Fabrication Approach for Multifunctional Graphene-based Thin Film Nano-composite Membranes with Enhanced Desalination and Antibacterial Characteristics.

A practical fabrication technique is presented to tackle the trade-off between the water flux and salt rejection of thin film composite (TFC) reverse osmosis (RO) membranes through controlled creation of a thinner active selective polyamide (PA) layer. The new thin film nano-composite (TFNC) RO membranes were synthesized with multifunctional poly tannic acid-functionalized graphene oxide nanosheets (pTA-f-GO) embedded in its PA thin active layer, which is produced through interfacial polymerization. The incorporation of pTA-f-GOL into the fabricated TFNC membranes resulted in a thinner PA layer with lower roughness and higher hydrophilicity compared to pristine membrane. These properties enhanced both the membrane water flux (improved by 40%) and salt rejection (increased by 8%) of the TFNC membrane. Furthermore, the incorporation of biocidal pTA-f-GO nanosheets into the PA active layer contributed to improving the antibacterial properties by 80%, compared to pristine membrane. The fabrication of the pTA-f-GO nanosheets embedded in the PA layer presented in this study is a very practical, scalable and generic process that can potentially be applied in different types of separation membranes resulting in less energy consumption, increased cost-efficiency and improved performance.

Bacterial degradation of microcystin toxins within a biologically active sand filter.

Microcystin toxins are a problem for water authorities as they are recalcitrant to conventional water treatment. In this study, biological sand filtration was assessed in laboratory column experiments for its ability to remove two microcystin analogues, microcystin-LR and microcystin-LA. A lag period of 3 days was evident prior to the commencement of degradation. Contact times were varied during the experiment; however, no microcystin was detected in the effluent after 4 days, even under conditions similar to those of a rapid sand filter. Removals of microcystin through the sand filters were shown to be primarily through biological degradation processes. Using polymerase chain reaction (PCR), biofilm, extracted from one of the sand filters that had effectively removed the microcystins, was shown to contain bacteria with the mlrA gene. Detection of this gene provided additional evidence that biological degradation of microcystin was the primary removal mechanism.