A qPCR-Based Tool to Diagnose the Presence of Harmful Cyanobacteria and Cyanotoxins in Drinking Water Sources.
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ABSTRACT: Harmful cyanobacteria have been an important concern for drinking water quality for quite some time, as they may produce cyanotoxins and odorants. Microcystis and Cylindrospermopsis are two common harmful cyanobacterial genera detected in freshwater lakes and reservoirs, with microcystins (MCs) and cylindrospermopsin (CYN) as their important metabolites, respectively. In this study, two sets of duplex qPCR systems were developed, one for quantifying potentially-toxigenic Microcystis and Microcystis, and the other one for cylindrospermopsin-producing cyanobacteria and Cylindrospermopsis. The duplex qPCR systems were developed and validated in the laboratory by using 338 samples collected from 29 reservoirs in Taiwan and her offshore islands. Results show that cell numbers of Microcystis and Cylindorspermopsis enumerated with microscopy, and MCs and CYN concentrations measured with the enzyme-linked immuno-sorbent assay method, correlated well with their corresponding gene copies determined with the qPCR systems (range of coefficients of determination R² = 0.392-0.740). The developed qPCR approach may serve as a useful tool for the water industry to diagnose the presence of harmful cyanobacteria and the potential presence of cyanotoxins in source waters.
Project description:Harmful cyanobacteria and their cyanotoxins may contaminate drinking water resources and their effective control remains challenging. The present study reports on cyanobacterial blooms and associated cyanotoxins in the Obrzyca River, a source of drinking water in Poland. The river was examined from July to October 2019 and concentrations of microcystins, anatoxin-a, and cylindrospermopsin were monitored. The toxicity of water samples was also tested using an ecotoxicological assay. All studied cyanotoxins were detected with microcystins revealing the highest levels. Maximal microcystin concentrations (3.97 μg/L) were determined in September at Uście point, exceeding the provisional guideline. Extracts from Uście point, where the dominant species were Dolichospermum flos-aquae (August), Microcystis aeruginosa (September), and Planktothrix agardhii (October), were toxic for Dugesia tigrina Girard. Microcystin concentrations (MC-LR and MC-RR) were positively correlated with cyanobacteria biovolume. Analysis of the chemical indicators of water quality has shown relationships between them and microcystins as well as cyanobacteria abundance.
Project description:Harmful cyanobacterial blooms have become increasingly common in freshwater ecosystems in recent decades, mainly due to eutrophication and climate change. Water becomes unreliable for human consumption. Here, we report a comprehensive study carried out to investigate the water quality of several Campina Grande reservoirs. Our approach included metagenomics, microbial abundance quantification, ELISA test for three cyanotoxins (microcystin, nodularins, and cylindrospermopsin), and in vivo ecotoxicological tests with zebrafish embryos. Cytometry analysis showed high cyanobacterial abundance, while metagenomics identified an average of 10.6% of cyanobacterial sequences, and demonstrated the presence of Microcystis, Cylindrospermopsis, and toxin coding genes in all ponds. Zebrafish embryos reared with pond water had high mortality and diverse malformations. Among the ponds analyzed, Araçagi showed the highest lethality (an average of 62.9 ± 0.8%), followed by Boqueirão (lethality average of 62.5 ± 0.8%). Here, we demonstrate that water from ponds undergoing extremely drought conditions have an abundance of potentially harmful cyanobacteria and their toxins. Our findings are consistent with a scenario in which polluted drinking water poses a great risk to human health.
Project description:Toxic cyanobacteria have been shown to accumulate in drinking water treatment plants that are susceptible to algal blooms. However, the risk for plants that do not experience algal blooms, but that receive a low influx of cells, is not well known. This study determined the extent of cell accumulation and presence of cyanotoxins across the treatment trains of four plants in the Great Lakes region. Samples were collected for microscopic enumeration and enzyme-linked immunosorbent assay (ELISA) measurements for microcystins, anatoxin-a, saxitoxin, cylindrospermopsin, and β-methylamino-L-alanine (BMAA). Low cell influxes (under 1000 cells/mL) resulted in significant cell accumulations (over 1 × 10⁵ cells/mL) in clarifier sludge and filter backwash samples. Microcystins peaked at 7.2 µg/L in one clarifier sludge sample, exceeding the raw water concentration by a factor of 12. Anatoxin-a was detected in the finished drinking water of one plant at 0.6 µg/L. BMAA may have been detected in three finished water samples, though inconsistencies among the BMAA ELISAs call these results into question. In summary, the results show that plants receiving a low influx of cells can be at risk of toxic cyanobacterial accumulation, and therefore, the absence of a bloom at the source does not indicate the absence of risk.
Project description:With increasing global water temperatures and nutrient runoff in recent decades, the blooming season of algae lasts longer, resulting in toxin concentrations that exceed safe limits for human consumption and for recreational use. From the different toxins, microcystin-LR has been reported as the main cyanotoxin related to liver cancer, and consequently its abundance in water is constantly monitored. In this work, we report a methodology for decorating cellulose nanofibrils with β-cyclodextrin or with poly(β-cyclodextrin) which were tested for the recovery of microcystin from synthetic water. The adsorption was followed by Quartz Crystal Microbalance with Dissipation monitoring (QCM-D), allowing for real-time monitoring of the adsorption behavior. A maximum recovery of 196 mg/g was obtained with the modified by cyclodextrin. Characterization of the modified substrate was confirmed with Fourier Transform Infrared Spectroscopy (FT-IR), X-ray Photoelectron Spectroscopy (XPS), Thermogravimetric Analysis (TGA), and Atomic Force Microscopy (AFM).
Project description:The prevalence of neurodegenerative disease (ND) is increasing, partly owing to extensions in lifespan, with a larger percentage of members living to an older age, but the ND aetiology and pathogenesis are not fully understood, and effective treatments are still lacking. Neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, and amyotrophic lateral sclerosis are generally thought to progress as a consequence of genetic susceptibility and environmental influences. Up to now, several environmental triggers have been associated with NDs, and recent studies suggest that some cyanotoxins, produced by cyanobacteria and acting through a variety of molecular mechanisms, are highly neurotoxic, although their roles in neuropathy and particularly in NDs are still controversial. In this review, we summarize the most relevant and recent evidence that points at cyanotoxins as environmental triggers in NDs development.
Project description:A data-driven approach to characterizing the risk of cyanobacteria-based harmful algal blooms (cyanoHABs) was undertaken for the Ohio River. Twenty-five years of river discharge data were used to develop Bayesian regression models that are currently applicable to 20 sites spread-out along the entire 1579 km of the river's length. Two site-level prediction models were developed based on the antecedent flow conditions of the two blooms that occurred on the river in 2015 and 2019: one predicts if the current year will have a bloom (the occurrence model), and another predicts bloom persistence (the persistence model). Predictors for both models were based on time-lagged average flow exceedances and a site's characteristic residence time under low flow conditions. Model results are presented in terms of probabilities of occurrence or persistence with uncertainty. Although the occurrence of the 2019 bloom was well predicted with the modeling approach, the limited number of events constrained formal model validation. However, as a measure of performance, leave-one-out cross validation returned low misclassification rates, suggesting that future years with flow time series like the previous bloom years will be correctly predicted and characterized for persistence potential. The prediction probabilities are served in real time as a component of a risk characterization tool/web application. In addition to presenting the model's results, the tool was designed with visualization options for studying water quality trends among eight river sites currently collecting data that could be associated with or indicative of bloom conditions. The tool is made accessible to river water quality professionals to support risk communication to stakeholders, as well as serving as a real-time water data monitoring utility.
Project description:Cyanotoxins are a diverse group of bioactive compounds produced by cyanobacteria that have adverse effects on human and animal health. While the phenomenon of cyanotoxin production in aquatic environments is well studied, research on cyanotoxins in terrestrial environments, where cyanobacteria abundantly occur in biocrusts, is still in its infancy. Here, we investigated the potential cyanotoxin production in cyanobacteria-dominated biological loess crusts (BLCs) from three different regions (China, Iran, and Serbia) and in cyanobacterial cultures isolated from the BLCs. The presence of cyanotoxins microcystins, cylindrospermopsin, saxitoxins, and β-N-methylamino-L-alanine was analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) method, while the presence of cyanotoxin-encoding genes (mcyE, cyrJ, sxtA, sxtG, sxtS, and anaC) was investigated by polymerase chain reaction (PCR) method. We could not detect any of the targeted cyanotoxins in the biocrusts or the cyanobacterial cultures, nor could we amplify any cyanotoxin-encoding genes in the cyanobacterial strains. The results are discussed in terms of the biological role of cyanotoxins, the application of cyanobacteria in land restoration programs, and the use of cyanotoxins as biosignatures of cyanobacterial populations in loess research. The article highlights the need to extend the field of research on cyanobacteria and cyanotoxin production to terrestrial environments.
Project description:Lithium (Li) is listed in the fifth Unregulated Contaminant Monitoring Rule (UCMR 5) because insufficient exposure data exists for lithium in drinking water. To help fill this data gap, lithium occurrence in source waters across the United States was assessed in 21 drinking water utilities. From the 369 samples collected from drinking water treatment plants (DWTPs), lithium ranged from 0.9 to 161 μg/L (median = 13.9 μg/L) in groundwater, and from <0.5 to 130 μg/L (median = 3.9 μg/L) in surface water. Lithium in 56% of the groundwater and 13% of the surface water samples were above non-regulatory Health-Based Screening Level (HBSL) of 10 μg/L. Sodium and lithium concentrations were strongly correlated: Kendall's τ > 0.6 (p < 0.001). As sodium is regularly monitored, this result shows that sodium can serve as an indicator to identify water sources at higher risk for elevated lithium. Lithium concentrations in the paired samples collected in source water and treated drinking water were almost identical showing lithium was not removed by conventional drinking water treatment processes. Additional sampling in wastewater effluents detected lithium at 0.8-98.2 μg/L (median = 9.9 μg/L), which suggests more research on impacts of lithium in direct and indirect potable reuse may be warranted, as the median was close to the HBSL. For comparison with the study samples collected from DWTPs, lithium concentrations from the national water quality portal (WQP) database were also investigated. Over 35,000 measurements were collected from waters that could potentially be used as drinking water sources (Cl- < 250 mg/L). Data from WQP had comparable median lithium concentrations: 18 and 20 μg/L for surface water and groundwater, respectively. Overall, this study provides a comprehensive occurrence potential for lithium in US drinking water sources and can inform the data collection effort in UCMR 5.
Project description:Maladaptive reward memories (MRMs) are involved in the development and maintenance of acquired overconsumption disorders, such as harmful alcohol and drug use. The process of memory reconsolidation - where stored memories become briefly labile upon retrieval - may offer a means to disrupt MRMs and prevent relapse. However, reliable means for pharmacologically weakening MRMs in humans remain elusive. Here we demonstrate that the N-methyl D-aspartate (NMDA) antagonist ketamine is able to disrupt MRMs in hazardous drinkers when administered immediately after their retrieval. MRM retrieval + ketamine (RET + KET) effectively reduced the reinforcing effects of alcohol and long-term drinking levels, compared to ketamine or retrieval alone. Blood concentrations of ketamine and its metabolites during the critical 'reconsolidation window' predicted beneficial changes only following MRM reactivation. Pharmacological reconsolidation interference may provide a means to rapidly rewrite maladaptive memory and should be further pursued in alcohol and drug use disorders.
Project description:Health-related concerns about cyanobacteria-laden sludge of drinking water treatment plants (DWTPs) have been raised in the past few years. Microscopic taxonomy, shotgun metagenomic sequencing, and microcystin (MC) measurement were applied to study the fate of cyanobacteria and cyanotoxins after controlled sludge storage (stagnation) in the dark in a full-scale drinking water treatment plant within 7 to 38 days. For four out of eight dates, cyanobacterial cell growth was observed by total taxonomic cell counts during sludge stagnation. The highest observed cell growth was 96% after 16 days of stagnation. Cell growth was dominated by potential MC producers such as Microcystis, Aphanocapsa, Chroococcus, and Dolichospermum. Shotgun metagenomic sequencing unveiled that stagnation stress shifts the cyanobacterial communities from the stress-sensitive Nostocales (e.g., Dolichospermum) order towards less compromised orders and potential MC producers such as Chroococcales (e.g., Microcystis) and Synechococcales (e.g., Synechococcus). The relative increase of cyanotoxin producers presents a health challenge when the supernatant of the stored sludge is recycled to the head of the DWTP or discharged into the source. These findings emphasize the importance of a strategy to manage cyanobacteria-laden sludge and suggest practical approaches should be adopted to control health/environmental impacts of cyanobacteria and cyanotoxins in sludge.