Persistence and Environmental Relevance of Extracellular Antibiotic Resistance Genes: Regulation by Nanoparticle Association

Chowdhury, Nadratun N.; Wiesner, Mark R.

doi:10.1089/ees.2021.0299

Cited by 7 publications

(13 citation statements)

References 49 publications

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“…Therefore, the ARGs detected on the 0.2 µm membrane are likely to represent ARGs adsorbed to nanoparticles or colloids. Experiments conducted by Chowdhury et al (2021) showed that this sorption of eARGs to nanoparticle surfaces is irreversible under specific conditions (humic-acid-functionalized silica nanoparticles) and to protects ARGs from DNAse I degradation ( Chowdhury and Wiesner, 2021 ). Moreover, DNA was found to adsorb better in sand and mineral surfaces, in general, under low pH conditions and higher chaotropic salt concentrations ( Lorenz and Wackernagel, 1987 ; Romanowski et al, 1991 ).…”

Section: Resultsmentioning

confidence: 99%

Cascade Filtration With PCR Detection and Field-Flow-Fractionation Online With ICP-MS for the Characterization of DNA Interaction With Suspended Particulate Matter

et al. 2022

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The variety of applied antibiotics in animal and human medicine results in the release, development, and spread of relevant numbers of antibiotic resistance genes (ARGs) in the environment. The majority of ARGs are present in intracellular forms (in bacteria). Neglected aspects are extracellular variants of ARGs (eARGs) and their fragments, which have been detected in surface-water samples and sediments. The stability of eARGs is expected to be low; however, binding to particulate matter is likely to improve their stability and also affect their transport and dissemination behavior. Few studies have investigated DNA particle interactions, mostly via indirect characterization of adduct formation in model systems but not in real environmental matrices. Therefore, our study aims at a novel approach for direct characterization of desoxyribonucleic acid (DNA) particle interactions using both cascade filtration and field-flow fractionation. Cascade filtration with quantitative polymerase chain reaction (qPCR) detection indicated retention of ARGs on filters with much larger pore sizes supporting the hypothesis of ARG-particle interactions. However, artifacts from membrane clogging or DNA–membrane interaction cannot be excluded. Consequently, asymmetric flow field-flow fractionation was investigated as an alternative separation technique with the advantage of particle separation in a thin channel, reducing the risk of artifacts. The key method parameters, membrane composition, molecular weight cut off, and carrier composition, were systematically investigated using a calf-thymus DNA-spiked surface-water sample as a model. The results clearly showed a shift in the elution time of clay particles suggesting the presence of DNA–clay adducts. Multi-element detection by inductively coupled plasma mass spectrometry (ICP-MS) enabled monitoring of clay via the Al, Fe, and Si signals and DNA via the P signal. Matching peak profiles for the new fraction in the fractograms of the ARG and DNA-spiked water sample support adduct formation. Further evidence was provided by a novel post-channel filtration approach for the separation of free DNA from DNA–clay adducts.

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Section: Resultsmentioning

confidence: 99%

Cascade Filtration With PCR Detection and Field-Flow-Fractionation Online With ICP-MS for the Characterization of DNA Interaction With Suspended Particulate Matter

et al. 2022

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“…eARGs were equilibrated with particles and DNase I prior to natural transformation assays. DNase I was added so that the impact of particle-mediated protection of genes could be assessed on transformation. The DNA sorption and DNase I exposure procedure is described in the Supporting Information (Text S5).…”

Section: Methodsmentioning

confidence: 99%

“…Extracted B. subtilis 1A189 DNA was exposed to a range of DNase I concentrations (0, 0.5, 1, 5, 10, and 20 μg/mL). Previously, it was reported that the amount of persistent eDNA varied significantly across this range of DNase I exposures . Therefore, this full range was tested here to evaluate how changes in persistence may correlate with changes in bioavailability.…”

Section: Methodsmentioning

confidence: 99%

“…The remaining eARGs (the 5 PCR amplified blt fragments, the single stranded 906 bp blt fragment, and DNA extracted from B. subtilis 1A354 and B. subtilis 1A491) were then prepared (exposed to a single DNase I concentration (1 μg/mL)) to determine impacts of eDNA properties on transformation. At this DNase I concentration, most free eDNA is fully degraded, ,, and remaining eDNA are likely particle-associated. Using this DNase I concentration ensures a comparison between the impact of gene properties on particle-free and particle-mediated transformation.…”

Section: Methodsmentioning

confidence: 99%

“…Among the particles that may associate with eDNA are naturally ubiquitous nanoparticles (NPs) which have been shown to irreversibly bind eARGs and protect them from enzymatic degradation . Based on these findings, this work hypothesizes that NPs may also mediate natural transformation.…”

Section: Introductionmentioning

confidence: 96%

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Investigating and Modeling the Regulation of Extracellular Antibiotic Resistance Gene Bioavailability by Naturally Occurring Nanoparticles

Chowdhury

Hicks

Wiesner

2022

Environ. Sci. Technol.

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Extracellular antibiotic resistance genes (eARGs) are widespread in the environment and can genetically transform bacteria. This work examined the role of environmentally relevant nanoparticles (NPs) in regulating eARG bioavailability. eARGs extracted from antibiotic-resistant B. subtilis were incubated with nonresistant recipient B. subtilis cells. In the mixture, particle type (either humic acid coated nanoparticles (HASNPs) or their micron-sized counterpart (HASPs)), DNase I concentration, and eARG type were systematically varied. Transformants were counted on selective media. Particles decreased bacterial growth and eARG bioavailability in systems without nuclease. When DNase I was present (≥5 μg/mL), particles increased transformation via chromosomal (but not plasmid-borne) eARGs. HASNPs increased transformation more than HASPs, indicating that the smaller nanoparticle with greater surface area per volume is more effective in increasing eARG bioavailability. These results were also modeled via particle aggregation theory, which represented eARG–bacteria interactions as transport leading to collision, followed by attachment. Using attachment efficiency as a fitting factor, the model predicted transformant concentrations within 35% of experimental data. These results confirm the ability of NPs to increase eARG bioavailability and suggest that particle aggregation theory may be a simplified and suitable framework to broadly predict eARG uptake.

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Removing antibiotic resistance genes under heavy metal stress with carbon-based materials and clay minerals: By sorption alone?

Huang

Huang²,

Chen³

et al. 2022

Chemical Engineering Journal

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Persistence and Environmental Relevance of Extracellular Antibiotic Resistance Genes: Regulation by Nanoparticle Association

Cited by 7 publications

References 49 publications

Cascade Filtration With PCR Detection and Field-Flow-Fractionation Online With ICP-MS for the Characterization of DNA Interaction With Suspended Particulate Matter

Cascade Filtration With PCR Detection and Field-Flow-Fractionation Online With ICP-MS for the Characterization of DNA Interaction With Suspended Particulate Matter

Investigating and Modeling the Regulation of Extracellular Antibiotic Resistance Gene Bioavailability by Naturally Occurring Nanoparticles

Removing antibiotic resistance genes under heavy metal stress with carbon-based materials and clay minerals: By sorption alone?

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