This study evaluated uranium sequestration performance in iron-rich (30 g/kg) sediment via biored... more This study evaluated uranium sequestration performance in iron-rich (30 g/kg) sediment via bioreduction followed by reoxidation. Field tests (1383 days) at Oak Ridge, Tennessee demonstrated that uranium contents in sediments increased after bioreduced sediments were re-exposed to nitrate and oxygen in contaminated groundwater. Bioreduction of contaminated sediments (1200 mg/kg U) with ethanol in microcosm reduced aqueous U from 0.37 to 0.023 mg/L. Aliquots of the bioreduced sediment were reoxidized with O2, H2O2, and NaNO3, respectively, over 285 days, resulting in aqueous U of 0.024, 1.58 and 14.4 mg/L at pH 6.30, 6.63 and 7.62, respectively. The source-and the three reoxidized sediments showed different desorption and adsorption behaviors of U, but all fit a Freundlich model. The adsorption capacities increased sharply at pH 4.5 to 5.5, plateaued at pH 5.5 to 7.0, then decreased sharply as pH increased from 7.0 to 8.0. The O2-reoxidized sediment retained a lower desorption efficiency at pH over 6.0. The NO3-reoxidized sediment exhibited higher adsorption capacity at pH 5.5 to 6.0. The pH-dependent adsorption onto Fe(III) oxides and formation of U coated particles and precipitates resulted in U sequestration, and bioreduction followed by reoxidation can enhance the U sequestration in sediment.
The polymerase chain reaction (PCR) is instrumental in molecular analysis of microorganisms, allo... more The polymerase chain reaction (PCR) is instrumental in molecular analysis of microorganisms, allowing for the selective amplification of nucleic acids directly from clinical and environmental samples. However, the principles that allow for targeted amplification of DNA become a hindrance when attempting to simultaneously discriminate and quantify complex mixtures of homologous genes. Here we present a simple solution to the quantitative problem by separating the enrichment and amplification aspects of a conventional PCR reaction. In this assay, genes are enriched using a DNA oligonucleotide capture probe and subsequently amplified in a two-step random amplification protocol. In order to evaluate the quantitative aspects of the gene capture assay, we used real-time quantitative-PCR to measure initial and final concentrations of homologous genes from constructed mixtures of genomes. Upon sampling for the universal DNA-dependent RNA polymerase gene, rpoC, we were able to demonstrate quantitative recoveries from a mixed DNA sample despite differences in gene copy number ranging up to 4 orders of magnitude. This suggests that minority populations as low as 0.01% of the total community are represented as accurately as populations at higher abundance. These results offer new possibilities for accurately and quantitatively monitoring diverse mixtures of microorganisms.
Struvite recovered from wastewater treatment plants can be upgraded into high-value, sustainable ... more Struvite recovered from wastewater treatment plants can be upgraded into high-value, sustainable fire retardants through encapsulation in viscoelastic fluids.
Environmental Science: Water Research & Technology
Heterotrophic nitrification is linked to the use of nitrogen-containing coagulants added to enhan... more Heterotrophic nitrification is linked to the use of nitrogen-containing coagulants added to enhance centrifugablity of biosolids.
Environmental Science: Water Research & Technology
First demonstration of the capture of salinity gradient energy from stormwater runoff to the ocea... more First demonstration of the capture of salinity gradient energy from stormwater runoff to the ocean, used to power UV-LED disinfection.
Recent studies have demonstrated the ability for polystyrene (PS) degradation within the gut of m... more Recent studies have demonstrated the ability for polystyrene (PS) degradation within the gut of mealworms ( Tenebrio molitor). To determine whether plastics may be broadly susceptible to biodegradation within mealworms, we evaluated the fate of polyethylene (PE) and mixtures (PE + PS). We find that PE biodegrades at comparable rates to PS. Mass balances indicate conversion of up 49.0 ± 1.4% of the ingested PE into a putative gas fraction (CO). The molecular weights ( M) of egested polymer residues decreased by 40.1 ± 8.5% in PE-fed mealworms and by 12.8 ± 3.1% in PS-fed mealworms. NMR and FTIR analyses revealed chemical modifications consistent with degradation and partial oxidation of the polymer. Mixtures likewise degraded. Our results are consistent with a nonspecific degradation mechanism. Analysis of the gut microbiome by next-generation sequencing revealed two OTUs ( Citrobacter sp. and Kosakonia sp.) strongly associated with both PE and PS as well as OTUs unique to each plast...
Applied and environmental microbiology, Jan 16, 2018
A site in Oak Ridge, Tennessee, USA, has sediments that contain >3% iron oxides and is contami... more A site in Oak Ridge, Tennessee, USA, has sediments that contain >3% iron oxides and is contaminated with uranium (U). The U(VI) was bioreduced to U(IV) and immobilizedthrough intermittent injections of ethanol. Then it was allowed to reoxidize via the invasion of low-pH (3.6-4.0), high-nitrate (up to 200 mM) groundwater back into the reduced zone for 1,383 days. To examine the biogeochemical response, high-throughput sequencing and network analysis were applied to characterize bacterial population shifts, as well as co-occurrence and co-exclusion patterns among microbial communities. Paired t-test indicated no significant changes of α-diversity for the bioactive wells. However, both nonmetric multidimensional scaling and analysis of similarity confirmed a significant distinction in the overall composition of the bacterial communities between the bioreduced and the reoxidized sediments. The top 20 major genera accounted for >70% of the cumulative contribution to the dissimilari...
Planning of water reuse systems is a complex endeavor. We have developed a software toolkit, IRIP... more Planning of water reuse systems is a complex endeavor. We have developed a software toolkit, IRIPT (Integrated Urban Reclaimed Water Infrastructure Planning Toolkit) that facilitates planning and design of reclaimed water infrastructure for both centralized and hybrid configurations that incorporate satellite treatment plants (STPs). The toolkit includes a Pipeline Designer (PRODOT) that optimizes routing and sizing of pipelines for wastewater capture and reclaimed water distribution, a Selector (SelWTP) that assembles and optimizes wastewater treatment trains, and a Calculator (CalcBenefit) that estimates fees, revenues, and subsidies of alternative designs. For hybrid configurations, a Locator (LocSTP) optimizes siting of STPs and associated wastewater diversions by identifying manhole locations where the flowrates are sufficient to ensure that wastewater extracted and treated at an adjacent STP can generate the revenue needed to pay for treatment and delivery to customers. Practi...
The United States Microbead-Free Waters Act was signed into law in December 2015. It is a biparti... more The United States Microbead-Free Waters Act was signed into law in December 2015. It is a bipartisan agreement that will eliminate one preventable source of microplastic pollution in the United States. Still, the bill is criticized for being too limited in scope, and also for discouraging the development of biodegradable alternatives that ultimately are needed to solve the bigger issue of plastics in the environment. Due to a lack of an acknowledged, appropriate standard for environmentally safe microplastics, the bill banned all plastic microbeads in selected cosmetic products. Here, we review the history of the legislation and how it relates to the issue of microplastic pollution in general, and we suggest a framework for a standard (which we call "Ecocyclable") that includes relative requirements related to toxicity, bioaccumulation, and degradation/assimilation into the natural carbon cycle. We suggest that such a standard will facilitate future regulation and legislation to reduce pollution while also encouraging innovation of sustainable technologies.
The first methanotrophic syntheses of polyhydroxyalkanoates (PHAs) that contain repeating units b... more The first methanotrophic syntheses of polyhydroxyalkanoates (PHAs) that contain repeating units beyond 3-hydroxybutyrate and 3-hydroxyvalerate are reported. New PHAs synthesized by methanotrophs include poly(3-hydroxybutyrate-co-4-hydroxybutyrate) (P(3HB-co-4HB)), poly(3-hydroxybutyrate-co-5-hydroxyvalerate-co-3-hydroxyvalerate) (P(3HB-co-5HV-co-3HV)), and poly(3-hydroxybutyrate-co-6-hydroxyhexanoate-co-4-hydroxybutyrate) (P(3HB-co-6HHx-co-4HB)). This was achieved from a pure culture of Methylocystis parvus OBBP where the primary substrate is methane and the corresponding ω-hydroxyalkanoate monomers are added as a co-substrate after the cells are subjected to nitrogen-limited conditions.
Kinetic models of anaerobic digestion (AD) are widely applied to soluble and particulate substrat... more Kinetic models of anaerobic digestion (AD) are widely applied to soluble and particulate substrates, but have not been systematically evaluated for bioplastics. Here, five models are evaluated to determine their suitability for modeling of anaerobic biodegradation of the bioplastic poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV): (1) first-order kinetics with and without a lag phase, (2) two-step first-order, (3) Monod (4) Contois, and (5) Gompertz. Three models that couple biomass growth with substrate hydrolysis (Monod, Contois, and Gompertz) gave the best overall fits for the data (R(2)>0.98), with reasonable estimates of ultimate CH4 production. The particle size limits of these models were then evaluated. Below a particle size of 0.8mm, rates of hydrolysis and acetogenesis exceeded rates of methanogenesis with accumulation of intermediates leading to a temporary inhibition of CH4 production. Based on model fit and simplicity, the Gompertz model is recommended for application...
Under iron-limiting conditions, Pseudomonas stutzeri KC secretes a small but as yet unidentified ... more Under iron-limiting conditions, Pseudomonas stutzeri KC secretes a small but as yet unidentified factor that transforms carbon tetrachloride (CT) to CO2 and nonvolatile products when activated by reduction at cell membranes. Pseudomonas fluorescens and other cell types activate the factor. Triparental mating was used to generate kanamycin-resistant lux::Tn5 recombinants of strain KC. Recombinants were streaked onto the surface of agar medium plugs in microtiter plates and were then screened for carbon tetrachloride degradation by exposing the plates to gaseous 14C-carbon tetrachloride. CT+ recombinants generated nonvolatile 14C-labeled products, but four CT- recombinants did not generate significant nonvolatile 14C-labeled products and had lost the ability to degrade carbon tetrachloride. When colonies of P. fluorescens were grown next to colonies of CT+ recombinants and were exposed to gaseous 14C-carbon tetrachloride, 14C-labeled products accumulated around the P. fluorescens colonies, indicating that the factor secreted by CT+ colonies had diffused through the agar and become activated. When P. fluorescens was grown next to CT- colonies, little carbon tetrachloride transformation was observed, indicating a lack of active factor. Expression of lux reporter genes in three of the CT- mutants was regulated by added iron and was induced under the same iron-limiting conditions that induce carbon tetrachloride transformation in the wild-type.
Oxidation and mobilization of microbially-generated U(IV) is of great concern for in situ uranium... more Oxidation and mobilization of microbially-generated U(IV) is of great concern for in situ uranium bioremediation. This study investigated the reoxidation of uranium by oxygen and nitrate in a sulfate-reducing enrichment and an iron-reducing enrichment derived from sediment and groundwater from the Field Research Center in Oak Ridge, Tennessee. Both enrichments were capable of reducing U(VI) rapidly. 16S rRNA gene clone libraries of the two enrichments revealed that Desulfovibrio spp. are dominant in the sulfate-reducing enrichment, and Clostridium spp. are dominant in the iron-reducing enrichment. In both the sulfate-reducing enrichment and the iron-reducing enrichment, oxygen reoxidized the previously reduced uranium but to a lesser extent in the iron-reducing enrichment. Moreover, in the iron-reducing enrichment, the reoxidized U(VI) was eventually re-reduced to its previous level. In both, the sulfate-reducing enrichment and the iron-reducing enrichment, uranium reoxidation did not occur in the presence of nitrate. The results indicate that the Clostridium-dominated iron-reducing communities created conditions that were more favorable for uranium stability with respect to reoxidation despite the fact that fewer electron equivalents were added to these systems. The likely reason is that more of the added electrons are present in a form that can reduce oxygen to water and U(VI) back to U(IV).
One method for the in situ remediation of uranium contaminated subsurface environments is the rem... more One method for the in situ remediation of uranium contaminated subsurface environments is the removal of highly soluble U(VI) from groundwater by microbial reduction to the sparingly soluble U(IV) mineral uraninite. Success of this remediation strategy will, in part, be determined by the extent and products of microbial reduction. In heterogeneous subsurface environments, microbial processes will likely yield a combination of U(IV) and U(VI) phases distributed throughout the soil matrix. Here, we use a combination of bulk X-ray absorption spectroscopy (XAS) and micro-focused XAS and X-ray diffraction to determine uranium speciation and distribution with sediment from a pilotscale uranium remediation project located in Oak Ridge, TN.
This study evaluated uranium sequestration performance in iron-rich (30 g/kg) sediment via biored... more This study evaluated uranium sequestration performance in iron-rich (30 g/kg) sediment via bioreduction followed by reoxidation. Field tests (1383 days) at Oak Ridge, Tennessee demonstrated that uranium contents in sediments increased after bioreduced sediments were re-exposed to nitrate and oxygen in contaminated groundwater. Bioreduction of contaminated sediments (1200 mg/kg U) with ethanol in microcosm reduced aqueous U from 0.37 to 0.023 mg/L. Aliquots of the bioreduced sediment were reoxidized with O2, H2O2, and NaNO3, respectively, over 285 days, resulting in aqueous U of 0.024, 1.58 and 14.4 mg/L at pH 6.30, 6.63 and 7.62, respectively. The source-and the three reoxidized sediments showed different desorption and adsorption behaviors of U, but all fit a Freundlich model. The adsorption capacities increased sharply at pH 4.5 to 5.5, plateaued at pH 5.5 to 7.0, then decreased sharply as pH increased from 7.0 to 8.0. The O2-reoxidized sediment retained a lower desorption efficiency at pH over 6.0. The NO3-reoxidized sediment exhibited higher adsorption capacity at pH 5.5 to 6.0. The pH-dependent adsorption onto Fe(III) oxides and formation of U coated particles and precipitates resulted in U sequestration, and bioreduction followed by reoxidation can enhance the U sequestration in sediment.
The polymerase chain reaction (PCR) is instrumental in molecular analysis of microorganisms, allo... more The polymerase chain reaction (PCR) is instrumental in molecular analysis of microorganisms, allowing for the selective amplification of nucleic acids directly from clinical and environmental samples. However, the principles that allow for targeted amplification of DNA become a hindrance when attempting to simultaneously discriminate and quantify complex mixtures of homologous genes. Here we present a simple solution to the quantitative problem by separating the enrichment and amplification aspects of a conventional PCR reaction. In this assay, genes are enriched using a DNA oligonucleotide capture probe and subsequently amplified in a two-step random amplification protocol. In order to evaluate the quantitative aspects of the gene capture assay, we used real-time quantitative-PCR to measure initial and final concentrations of homologous genes from constructed mixtures of genomes. Upon sampling for the universal DNA-dependent RNA polymerase gene, rpoC, we were able to demonstrate quantitative recoveries from a mixed DNA sample despite differences in gene copy number ranging up to 4 orders of magnitude. This suggests that minority populations as low as 0.01% of the total community are represented as accurately as populations at higher abundance. These results offer new possibilities for accurately and quantitatively monitoring diverse mixtures of microorganisms.
Struvite recovered from wastewater treatment plants can be upgraded into high-value, sustainable ... more Struvite recovered from wastewater treatment plants can be upgraded into high-value, sustainable fire retardants through encapsulation in viscoelastic fluids.
Environmental Science: Water Research & Technology
Heterotrophic nitrification is linked to the use of nitrogen-containing coagulants added to enhan... more Heterotrophic nitrification is linked to the use of nitrogen-containing coagulants added to enhance centrifugablity of biosolids.
Environmental Science: Water Research & Technology
First demonstration of the capture of salinity gradient energy from stormwater runoff to the ocea... more First demonstration of the capture of salinity gradient energy from stormwater runoff to the ocean, used to power UV-LED disinfection.
Recent studies have demonstrated the ability for polystyrene (PS) degradation within the gut of m... more Recent studies have demonstrated the ability for polystyrene (PS) degradation within the gut of mealworms ( Tenebrio molitor). To determine whether plastics may be broadly susceptible to biodegradation within mealworms, we evaluated the fate of polyethylene (PE) and mixtures (PE + PS). We find that PE biodegrades at comparable rates to PS. Mass balances indicate conversion of up 49.0 ± 1.4% of the ingested PE into a putative gas fraction (CO). The molecular weights ( M) of egested polymer residues decreased by 40.1 ± 8.5% in PE-fed mealworms and by 12.8 ± 3.1% in PS-fed mealworms. NMR and FTIR analyses revealed chemical modifications consistent with degradation and partial oxidation of the polymer. Mixtures likewise degraded. Our results are consistent with a nonspecific degradation mechanism. Analysis of the gut microbiome by next-generation sequencing revealed two OTUs ( Citrobacter sp. and Kosakonia sp.) strongly associated with both PE and PS as well as OTUs unique to each plast...
Applied and environmental microbiology, Jan 16, 2018
A site in Oak Ridge, Tennessee, USA, has sediments that contain >3% iron oxides and is contami... more A site in Oak Ridge, Tennessee, USA, has sediments that contain >3% iron oxides and is contaminated with uranium (U). The U(VI) was bioreduced to U(IV) and immobilizedthrough intermittent injections of ethanol. Then it was allowed to reoxidize via the invasion of low-pH (3.6-4.0), high-nitrate (up to 200 mM) groundwater back into the reduced zone for 1,383 days. To examine the biogeochemical response, high-throughput sequencing and network analysis were applied to characterize bacterial population shifts, as well as co-occurrence and co-exclusion patterns among microbial communities. Paired t-test indicated no significant changes of α-diversity for the bioactive wells. However, both nonmetric multidimensional scaling and analysis of similarity confirmed a significant distinction in the overall composition of the bacterial communities between the bioreduced and the reoxidized sediments. The top 20 major genera accounted for >70% of the cumulative contribution to the dissimilari...
Planning of water reuse systems is a complex endeavor. We have developed a software toolkit, IRIP... more Planning of water reuse systems is a complex endeavor. We have developed a software toolkit, IRIPT (Integrated Urban Reclaimed Water Infrastructure Planning Toolkit) that facilitates planning and design of reclaimed water infrastructure for both centralized and hybrid configurations that incorporate satellite treatment plants (STPs). The toolkit includes a Pipeline Designer (PRODOT) that optimizes routing and sizing of pipelines for wastewater capture and reclaimed water distribution, a Selector (SelWTP) that assembles and optimizes wastewater treatment trains, and a Calculator (CalcBenefit) that estimates fees, revenues, and subsidies of alternative designs. For hybrid configurations, a Locator (LocSTP) optimizes siting of STPs and associated wastewater diversions by identifying manhole locations where the flowrates are sufficient to ensure that wastewater extracted and treated at an adjacent STP can generate the revenue needed to pay for treatment and delivery to customers. Practi...
The United States Microbead-Free Waters Act was signed into law in December 2015. It is a biparti... more The United States Microbead-Free Waters Act was signed into law in December 2015. It is a bipartisan agreement that will eliminate one preventable source of microplastic pollution in the United States. Still, the bill is criticized for being too limited in scope, and also for discouraging the development of biodegradable alternatives that ultimately are needed to solve the bigger issue of plastics in the environment. Due to a lack of an acknowledged, appropriate standard for environmentally safe microplastics, the bill banned all plastic microbeads in selected cosmetic products. Here, we review the history of the legislation and how it relates to the issue of microplastic pollution in general, and we suggest a framework for a standard (which we call "Ecocyclable") that includes relative requirements related to toxicity, bioaccumulation, and degradation/assimilation into the natural carbon cycle. We suggest that such a standard will facilitate future regulation and legislation to reduce pollution while also encouraging innovation of sustainable technologies.
The first methanotrophic syntheses of polyhydroxyalkanoates (PHAs) that contain repeating units b... more The first methanotrophic syntheses of polyhydroxyalkanoates (PHAs) that contain repeating units beyond 3-hydroxybutyrate and 3-hydroxyvalerate are reported. New PHAs synthesized by methanotrophs include poly(3-hydroxybutyrate-co-4-hydroxybutyrate) (P(3HB-co-4HB)), poly(3-hydroxybutyrate-co-5-hydroxyvalerate-co-3-hydroxyvalerate) (P(3HB-co-5HV-co-3HV)), and poly(3-hydroxybutyrate-co-6-hydroxyhexanoate-co-4-hydroxybutyrate) (P(3HB-co-6HHx-co-4HB)). This was achieved from a pure culture of Methylocystis parvus OBBP where the primary substrate is methane and the corresponding ω-hydroxyalkanoate monomers are added as a co-substrate after the cells are subjected to nitrogen-limited conditions.
Kinetic models of anaerobic digestion (AD) are widely applied to soluble and particulate substrat... more Kinetic models of anaerobic digestion (AD) are widely applied to soluble and particulate substrates, but have not been systematically evaluated for bioplastics. Here, five models are evaluated to determine their suitability for modeling of anaerobic biodegradation of the bioplastic poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV): (1) first-order kinetics with and without a lag phase, (2) two-step first-order, (3) Monod (4) Contois, and (5) Gompertz. Three models that couple biomass growth with substrate hydrolysis (Monod, Contois, and Gompertz) gave the best overall fits for the data (R(2)>0.98), with reasonable estimates of ultimate CH4 production. The particle size limits of these models were then evaluated. Below a particle size of 0.8mm, rates of hydrolysis and acetogenesis exceeded rates of methanogenesis with accumulation of intermediates leading to a temporary inhibition of CH4 production. Based on model fit and simplicity, the Gompertz model is recommended for application...
Under iron-limiting conditions, Pseudomonas stutzeri KC secretes a small but as yet unidentified ... more Under iron-limiting conditions, Pseudomonas stutzeri KC secretes a small but as yet unidentified factor that transforms carbon tetrachloride (CT) to CO2 and nonvolatile products when activated by reduction at cell membranes. Pseudomonas fluorescens and other cell types activate the factor. Triparental mating was used to generate kanamycin-resistant lux::Tn5 recombinants of strain KC. Recombinants were streaked onto the surface of agar medium plugs in microtiter plates and were then screened for carbon tetrachloride degradation by exposing the plates to gaseous 14C-carbon tetrachloride. CT+ recombinants generated nonvolatile 14C-labeled products, but four CT- recombinants did not generate significant nonvolatile 14C-labeled products and had lost the ability to degrade carbon tetrachloride. When colonies of P. fluorescens were grown next to colonies of CT+ recombinants and were exposed to gaseous 14C-carbon tetrachloride, 14C-labeled products accumulated around the P. fluorescens colonies, indicating that the factor secreted by CT+ colonies had diffused through the agar and become activated. When P. fluorescens was grown next to CT- colonies, little carbon tetrachloride transformation was observed, indicating a lack of active factor. Expression of lux reporter genes in three of the CT- mutants was regulated by added iron and was induced under the same iron-limiting conditions that induce carbon tetrachloride transformation in the wild-type.
Oxidation and mobilization of microbially-generated U(IV) is of great concern for in situ uranium... more Oxidation and mobilization of microbially-generated U(IV) is of great concern for in situ uranium bioremediation. This study investigated the reoxidation of uranium by oxygen and nitrate in a sulfate-reducing enrichment and an iron-reducing enrichment derived from sediment and groundwater from the Field Research Center in Oak Ridge, Tennessee. Both enrichments were capable of reducing U(VI) rapidly. 16S rRNA gene clone libraries of the two enrichments revealed that Desulfovibrio spp. are dominant in the sulfate-reducing enrichment, and Clostridium spp. are dominant in the iron-reducing enrichment. In both the sulfate-reducing enrichment and the iron-reducing enrichment, oxygen reoxidized the previously reduced uranium but to a lesser extent in the iron-reducing enrichment. Moreover, in the iron-reducing enrichment, the reoxidized U(VI) was eventually re-reduced to its previous level. In both, the sulfate-reducing enrichment and the iron-reducing enrichment, uranium reoxidation did not occur in the presence of nitrate. The results indicate that the Clostridium-dominated iron-reducing communities created conditions that were more favorable for uranium stability with respect to reoxidation despite the fact that fewer electron equivalents were added to these systems. The likely reason is that more of the added electrons are present in a form that can reduce oxygen to water and U(VI) back to U(IV).
One method for the in situ remediation of uranium contaminated subsurface environments is the rem... more One method for the in situ remediation of uranium contaminated subsurface environments is the removal of highly soluble U(VI) from groundwater by microbial reduction to the sparingly soluble U(IV) mineral uraninite. Success of this remediation strategy will, in part, be determined by the extent and products of microbial reduction. In heterogeneous subsurface environments, microbial processes will likely yield a combination of U(IV) and U(VI) phases distributed throughout the soil matrix. Here, we use a combination of bulk X-ray absorption spectroscopy (XAS) and micro-focused XAS and X-ray diffraction to determine uranium speciation and distribution with sediment from a pilotscale uranium remediation project located in Oak Ridge, TN.
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Papers by Craig Criddle