Water tracking in surface water, groundwater and soils using free and alginate-chitosan encapsulated synthetic DNA tracers

Pang, Liping; Abeysekera, Gayan; Hanning, Kyrin R.; Premaratne, Aruni; Robson, Beth; Abraham, Phillip; Sutton, Richard; Hanson, Carl; Hadfield, John; Heiligenthal, Laura; Stone, Dana; McBeth, Kurt; Billington, Craig

doi:10.1016/j.watres.2020.116192

Cited by 27 publications

(46 citation statements)

References 37 publications

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“…To minimize the impacts of surrogates on humans and the eco-environment, biodegradable and biocompatible materials should be used for surrogate synthesis if surrogates are to be applied to the EWS, including drinking water distribution systems, and eco-sensitive freshwaters. Naturally occurring biopolymers can be used for this purpose, and recently, alginate and chitosan have been utilized to generate microencapsulated synthetic DNA tracers for water applications …”

Section: Introductionmentioning

confidence: 99%

Surface-Modified Biopolymer Microparticles: A Potential Surrogate for Studying Legionella pneumophila Attachment to Biofilms in Engineered Water Systems

et al. 2021

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Despite numerous legionellosis outbreaks attributed to contaminated engineered water systems (EWSs), no suitable surrogate has been established for studying Legionella pneumophila in the EWSs. In this study, we developed a novel biopolymer surrogate by using DNA-encapsulated, amino acid-modified alginate−calcium carbonate microparticles. The surrogate mimicked L. pneumophila's size, rod shape, surface charge, and hydrophobicity. Preliminary proof-of-concept validations were conducted with the surrogate and L. pneumophila in laboratory flow-through biofilm bioreactor experiments with and without chlorine. The 2 entities displayed similar attachment/detachment magnitudes and kinetics to/from Pseudomonas f luorescens biofilms grown on stainless steel piping material. Their relative concentrations and peak attachment values were within the same orders of magnitude, and both exhibited slower attachment than detachment processes. Compared with the experiments without chlorine, surrogate and L. pneumophila biofilm attachment was significantly reduced in the presence of chlorine. P. f luorescens biofilm formation was not affected by exposure to the surrogate microparticles that were made of biocompatible materials. The promising results from our study suggest that further validations of this new surrogate are warranted. It may create new pathways for investigating L. pneumophila mobility and persistence in EWSs, which will facilitate better control of legionellosis risks.

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Section: Introductionmentioning

confidence: 99%

Surface-Modified Biopolymer Microparticles: A Potential Surrogate for Studying Legionella pneumophila Attachment to Biofilms in Engineered Water Systems

et al. 2021

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“…Sequences of the primers used in this study are listed in Supplemental Table S1. BiADO primers amplify 279 bp of the 16S gene from Bifidobacterium adolescentis, which is a widely used indicator of fecal pollution (Matsuki et al, 2004), and K9 primers amplify 352 bp of a synthetic construct cloned into pcloneEZ, which we have developed for tracking water contamination (Pang et al, 2020). There are two forward primers used for BiADO detection, to enable the detection of both genotypes A and B (Matsuki et al, 2004).…”

Section: Instruments Templates and Primersmentioning

confidence: 99%

“…We chose these targets because K9 is one of the DNA tracers we have developed for tracking water contamination (Pang et al, 2020), and BiADO is a widely used indicator of fecal pollution (Matsuki et al, 2004). Results of these SYBR green qPCR assays are shown in Figure 1.…”

Section: Standard Curvesmentioning

confidence: 99%

“…Notably, the gold-standard test for the SARS-CoV-2 virus, which is the cause of the current COVID-19 global pandemic, is a reverse-transcriptase qPCR test (van Kasteren et al, 2020). Quantitative real-time polymerase chain reaction is a key technique our Institute uses for environmental monitoring including tracking water pollution using DNA tracers (Pang et al, 2020), using surrogates for evaluation of pathogen transport (Pang et al, 2014) and environmental fecal source tracking (Devane et al, 2019). However, one significant limitation of qPCR technology for field research has been the lack of portability of the amplification and detection equipment needed to perform qPCR assays, necessitating transport and often storage of samples at the laboratory prior to analyses.…”

Section: Introductionmentioning

confidence: 99%

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Utility of a field deployable qPCR instrument for analyzing freshwater quality

Billington

Abeysekera

Scholes

et al. 2021

Agrosystems Geosci & Env

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A significant limitation of quantitative real-time polymerase chain reaction (qPCR) technology for environmental research has been the lack of portability of the amplification and detection equipment needed to perform qPCR assays in the field. The ability to perform qPCR assays in the field would significantly improve the utility of the technology, enabling quicker risk management decisions in acute environmental emergencies, and speeding the pace of environmental research. We tested a portable qPCR instrument, Liberty16, to evaluate if this instrument would be useful for qPCR analysis in our fieldwork. Parallel testing of the Liberty16 with our existing LightCycler 480 Instrument II revealed similar repeatability and sensitivity for the detection of targets in known and blind freshwater samples by qPCR. Both instruments were able to detect all target sequences at 20 copies per reaction and up to 66% targets in a 10-fold dilution. In seven of eight blind freshwater samples, results from both instruments were consistent for detected or nondetected target. In the other sample, the target was only detected by the Liberty16. Portable instruments of this type may open up new avenues for researchers to perform qPCR analyses in the field.

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“…The main criteria for biopolymer selection were solubility in water at ambient temperature, overall surface charge, availability of facile methods for microparticle synthesis without the requirement of specialized equipment or hazardous chemicals for potential future upscaling and environmental application, and the availability of functional groups for surface modifications. In addition, the mimics needed to incorporate short DNA tracer sequences to enable their tracking in the environment [15]. Synthesis techniques were evaluated in the laboratory to obtain a rod-shaped biopolymer surrogate with a size, surface charge, and relative hydrophobicity similar to stationary-phase L. pneumophila.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of Biopolymer Materials and Synthesis Techniques to Develop a Rod-Shaped Biopolymer Surrogate for Legionella pneumophila

et al. 2022

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Biopolymer microparticles have been developed for applications that require biocompatibility and biodegradability, such as drug delivery. In this study, we assessed the production of microparticles using carnauba wax, κ-carrageenan, alginate, and poly (lactic-co-glycolic acid) (PLGA) with the aim of developing a novel, DNA-tracer-loaded, biopolymer surrogate with a size, shape, surface charge, and relative hydrophobicity similar to stationary-phase Legionella pneumophila to mimic the bacteria’s mobility and persistence in engineered water systems. We found that the type and concentration of biopolymer, reaction conditions, and synthesis methods affected the morphology, surface charge, relative hydrophobicity, and DNA tracer loading efficiency of the biopolymer microparticles produced. Carnauba wax, κ-carrageenan, and alginate (Protanal®, and low and medium viscosity) produced highly polydisperse microspheres. In contrast, PLGA and alginate-CaCO3 produced uniform microspheres and rod-shaped microparticles, respectively, with high DNA tracer loading efficiencies (PLGA 70% and alginate-CaCO3 95.2 ± 5.7%) and high reproducibilities. Their synthesis reproducibility was relatively high. The relative hydrophobicity of PLGA microspheres closely matched the cell surface hydrophobicity of L. pneumophila but not the bacterial morphology, whereas the polyelectrolyte layer-by-layer assembly was required to enhance the relative hydrophobicity of alginate-CaCO3 microparticles. Following this surface modification, alginate-CaCO3 microparticles represented the best match to L. pneumophila in size, morphology, surface charge, and relative hydrophobicity. This new biopolymer surrogate has the potential to be used as a mimic to study the mobility and persistence of L. pneumophila in water systems where the use of the pathogen is impractical and unsafe.

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Water tracking in surface water, groundwater and soils using free and alginate-chitosan encapsulated synthetic DNA tracers

Cited by 27 publications

References 37 publications

Surface-Modified Biopolymer Microparticles: A Potential Surrogate for Studying Legionella pneumophila Attachment to Biofilms in Engineered Water Systems

Surface-Modified Biopolymer Microparticles: A Potential Surrogate for Studying Legionella pneumophila Attachment to Biofilms in Engineered Water Systems

Utility of a field deployable qPCR instrument for analyzing freshwater quality

Evaluation of Biopolymer Materials and Synthesis Techniques to Develop a Rod-Shaped Biopolymer Surrogate for Legionella pneumophila

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