Paul Hatzinger - Academia.edu (original) (raw)

Papers by Paul Hatzinger

Research paper thumbnail of Evaluation of microbial transport during aerobic bioaugmentation of an RDX-contaminated aquifer

Biodegradation, 2015

In situ bioaugmentation with aerobic hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX)-degrading bact... more In situ bioaugmentation with aerobic hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX)-degrading bacteria is being considered for treatment of explosives-contaminated groundwater at Umatilla Chemical Depot, Oregon (UMCD). Two forced-gradient bacterial transport tests of site groundwater containing chloride or bromide tracer and either a mixed culture of Gordonia sp. KTR9 (xplA (+)Km(R)), Rhodococcus jostii RHA1 (pGKT2 transconjugant; xplA (+)Km(R)) and Pseudomonas fluorescens I-C (xenB (+)), or a single culture of Gordonia sp. KTR9 (xplA (+); i.e. wild-type) were conducted at UMCD. Groundwater monitoring evaluated cell viability and migration in the injection well and downgradient monitoring wells. Enhanced degradation of RDX was not evaluated in these demonstrations. Quantitative PCR analysis of xplA, the kanamycin resistance gene (aph), and xenB indicated that the mixed culture was transported at least 3 m within 2 h of injection. During a subsequent field injection of bioaugmented groundwater, strain KTR9 (wild-type) migrated up to 23-m downgradient of the injection well within 3 days. Thus, the three RDX-degrading strains were effectively introduced and transported within the UMCD aquifer. This demonstration represents an innovative application of bioaugmentation to potentially enhance RDX biodegradation in aerobic aquifers.

Research paper thumbnail of Effect of Aging on the Biodegradation of Chemicals

Research paper thumbnail of Overview of in situ and ex situ bioremediation approaches for perchlorate

A mathematical model was developed to describe the biodegradation kinetics of perchlorate in the ... more A mathematical model was developed to describe the biodegradation kinetics of perchlorate in the presence of nitrate and oxygen as competing electron acceptors. The rate of perchlorate degradation is described as a function of the electron donor (acetate) degradation rate, the concentration of the alternate electron acceptors, and rates of biomass growth and decay. The kinetics of biomass growth are described using a modified Monod model, and inhibition factors are incorporated to describe the influence of oxygen and nitrate on perchlorate degradation. In order to develop input parameters for the model, a series of batch biodegradation studies were performed using Azospira suillum JPLRND, a perchlorate-degrading strain isolated from groundwater. This strain is capable of utilizing oxygen, nitrate, or perchlorate as terminal electron acceptors. The maximum specific growth rate (µ max ) and half-saturation constant (K don S ) for the bacterium when utilizing either perchlorate or nitrate were similar; 0.16 per h and 158 mg acetate/L, respectively. However, these parameters were different when the strain was growing on oxygen. In this case, µ max and K don S were 0.22 per h and 119 mg acetate/L, respectively. The batch experiments also revealed that nitrate inhibits perchlorate biodegradation by this strain. This finding was incorporated into the model by applying an inhibition coefficient (K nit i ) value of 25 mg nitrate/L. Combined with appropriate groundwater transport models, this model can be used to predict perchlorate biodegradation during in situ remediation efforts.

Research paper thumbnail of Using Stable Isotope Ratio Analysis to Distinguish Perchlorate Sources

Research paper thumbnail of ESTCP Project ER-200829

Research paper thumbnail of Mapping groundwater recharge areas for land use planning

Research paper thumbnail of Perchlorate Biodegradation for Water Treatment

Environmental Science & Technology, 2005

Research paper thumbnail of Rat kidney proximal tubule cells in defined medium: the roles of cholera toxin, extracellular calcium and serum in cell growth and expression of gamma-glutamyltransferase

In Vitro Cellular & Developmental Biology

A culture system is described in which rat kidney proximal tubule epithelial cells {RPTE) can be ... more A culture system is described in which rat kidney proximal tubule epithelial cells {RPTE) can be prepared with good yield and high viability and grown in culture under serum-free conditions. The cells require EGF, insulin, cholera toxin and either l% dialyzed serum or a complex of bovine serum albumin with oleic acid (BSA/OAk The cells can be maintained for long periods of time and express several markers for RPTE. The cells have both alkaline phosphatase and ?-glutamyltransferase activity and respond to parathyroid hormone but not vasopressin. The specific activity of y-glutamyltransferase decreases when the cells begin to grow, but increases when they reach confluence. Extracellular calcium plays a role in the induction of y-glutamyltransferase in confluent cells. Cells grown in media containing low calcium, i.e. less than 0.4 mM, have reduced specific activity of ),-glutamyltransferase. Extracellular calcium also alters the morphology of the cells in that cells grown in low calcium are single cells or loose clusters suggesting poor cell-cell contact. When the calcium is raised to 1.0 mM, the cells change their shape and organization to adopt the morphology of cells maintained continuously in 1.0 mM calcium. The cells can be passaged onto plastic surfaces which have been coated with collagen but cannot be subcultured on uncoated or serum coated plastic. This culture system will be a useful model for the investigation of renal carcinogenesis and the role of cell proliferation in that process. This paper reports a method for isolation and, uniquely, multipassage culture of rat proximal tubule epithelial cells. The authors use this culture system to explore some aspects of the physiology of these cells, demonstrating the value of the model.

Research paper thumbnail of Quantitation of multiple pathways for the metabolism of nephrotoxic cysteine conjugates using selective inhibitors of L-α-hydroxy acid oxidase (L-amino oxidase) and cysteine conjugate β-lyase

Drug Metabolism and Disposition

In this study, we have established the selectivity of inhibitors for rat kidney cysteine conjugat... more In this study, we have established the selectivity of inhibitors for rat kidney cysteine conjugate beta-lyase and L-alpha-hydroxy acid oxidase (L-amino acid oxidase) and have used these inhibitors to explore the relative roles of these two enzymes in the metabolism of nephrotoxic cysteine conjugates by rat kidney homogenate. In addition, we have investigated the relationship between structure and the metabolism of toxic cysteine conjugates by purified rat kidney L-alpha-hydroxy acid oxidase. With purified enzyme, S-(1,2,3,4,4-pentachlorobutadienyl)-L-cysteine (PCBDC) was about four times more active than S(1,2-dichlorovinyl)-L-cysteine (DCVC). Three alkyl conjugates were less active than DCVC. Purified L-alpha-hydroxy acid oxidase was not inhibited by the beta-lyase inhibitor aminooxyacetic acid but was inactivated by 2-hydroxy-3-butynoate. PCBDC metabolism in rat kidney homogenate was inhibited 74% by aminooxyacetic acid and 42% by 2-hydroxy-3-butynoate, whereas DCVC metabolism was inhibited 77% by aminooxyacetic acid and 28% by 2-hydroxy-3-butynoate. However, only aminooxyacetic acid inhibited the binding of 35S label from [35S]DCVC. Based on these results we have reached three conclusions. First, L-alpha-hydroxy acid oxidase plays a significant role in the metabolism of some cysteine conjugates. Second, metabolism of DCVC by L-alpha-hydroxy acid oxidase does not contribute directly to covalent binding. Third, as much as 65% of DCVC may be metabolized to its corresponding alpha-keto acid. The results are discussed with regard to the nephrotoxicity of cysteine conjugates.

Research paper thumbnail of Perchlorate and nitrate in situ bioremediation of ground water

Research paper thumbnail of Modeling In Situ Bioremediation of Perchlorate-Contaminated Groundwater

Perchlorate-contaminated groundwater is a significant national problem. An innovative technology ... more Perchlorate-contaminated groundwater is a significant national problem. An innovative technology was recently developed which uses a pair of dual-screened treatment wells to mix an electron donor into perchlorate- contaminated groundwater in order to effect in situ bioremediation of the perchlorate by indigenous perchlorate reducing bacteria (PRB) without the need to extract the contaminated water from the subsurface. The two treatment

Research paper thumbnail of In situ assessment of the transport and microbial consumption of oxygen in ground water, Cape Cod, Massachusetts

Research paper thumbnail of Guidance manual for forensic analysis of perchlorate in groundwater using chlorine and oxygen isotopic analyses

Research paper thumbnail of Field Demonstration of In Situ Perchlorate Bioremediation in Groundwater

Research paper thumbnail of Application of (13)C and (15)N stable isotope probing to characterize RDX degrading microbial communities under different electron-accepting conditions

Journal of hazardous materials, Jan 22, 2015

This study identified microorganisms capable of using the explosive hexahydro-1,3,5-trinitro-1,3,... more This study identified microorganisms capable of using the explosive hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) or its metabolites as carbon and/or nitrogen sources under different electron-accepting conditions using (13)C and (15)N stable isotope probing (SIP). Mesocosms were constructed using groundwater and aquifer solids from an RDX-contaminated aquifer. The mesocosms received succinate as a carbon source and one of four electron acceptors (nitrate, manganese(IV), iron(III), or sulfate) or no additional electron acceptor (to stimulate methanogenesis). When RDX degradation was observed, subsamples from each mesocosm were removed and amended with (13)C3- or ring-(15)N3-, nitro-(15)N3-, or fully-labeled (15)N6-RDX, followed by additional incubation and isolation of labeled nucleic acids. A total of fifteen 16S rRNA sequences, clustering in α- and γ-Proteobacteria, Clostridia, and Actinobacteria, were detected in the (13)C-DNA fractions. A total of twenty seven sequences were deri...

Research paper thumbnail of Factors affecting sequestration and bioavailability of phenanthrene in soils

Environmental Toxicology and Chemistry

A study was conducted to determine factors affecting the sequestration and changes in bioavailabi... more A study was conducted to determine factors affecting the sequestration and changes in bioavailability as phenanthrene persists in soils. Phenanthrene became sequestered in seven soils differing appreciably in organic matter and clay content as measured by earthworm uptake, bacterial mineralization, or extractability. Phenanthrene also became sequestered as it aged in soil aggregates of various sizes as measured by decline in availability to a bacterium, a mild extractant, or both. Wetting and drying a soil during aging reduced the amount of phenanthrene recovered by a mild extractant and the rate and extent of bacterial mineralization of the hydrocarbon. After biodegradation of phenanthrene added to the soil, more of the compound remained if it had been aged than if it had not been aged. Wetting and drying the soil during aging further increased the amount of phenanthrene remaining after biodegradation. The rate and extent of bacterial mineralization of phenanthrene were less in lea...

Research paper thumbnail of In-Situ and Ex-Situ Bioremediation Options for Treating Perchlorate in Groundwater

Remediation Journal, 2002

Research paper thumbnail of Active Bioremediation

SERDP/ESTCP Environmental Remediation Technology, 2009

Research paper thumbnail of Isotopic Tracing of Perchlorate in the Environment

Handbook of Environmental Isotope Geochemistry, 2011

Isotopic measurements can be used for tracing the sources and behavior of environmental contamina... more Isotopic measurements can be used for tracing the sources and behavior of environmental contaminants. Perchlorate (ClO 4 À ) has been detected

Research paper thumbnail of Applications of Selective Ion Exchange for Perchlorate Removal, Recovery and Environmental Forensics

Ion Exchange and Solvent Extraction Series, 2011

Isotopic measurements can be used for tracing the sources and behavior of environmental contamina... more Isotopic measurements can be used for tracing the sources and behavior of environmental contaminants. Perchlorate (ClO 4 À ) has been detected

Research paper thumbnail of Evaluation of microbial transport during aerobic bioaugmentation of an RDX-contaminated aquifer

Biodegradation, 2015

In situ bioaugmentation with aerobic hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX)-degrading bact... more In situ bioaugmentation with aerobic hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX)-degrading bacteria is being considered for treatment of explosives-contaminated groundwater at Umatilla Chemical Depot, Oregon (UMCD). Two forced-gradient bacterial transport tests of site groundwater containing chloride or bromide tracer and either a mixed culture of Gordonia sp. KTR9 (xplA (+)Km(R)), Rhodococcus jostii RHA1 (pGKT2 transconjugant; xplA (+)Km(R)) and Pseudomonas fluorescens I-C (xenB (+)), or a single culture of Gordonia sp. KTR9 (xplA (+); i.e. wild-type) were conducted at UMCD. Groundwater monitoring evaluated cell viability and migration in the injection well and downgradient monitoring wells. Enhanced degradation of RDX was not evaluated in these demonstrations. Quantitative PCR analysis of xplA, the kanamycin resistance gene (aph), and xenB indicated that the mixed culture was transported at least 3 m within 2 h of injection. During a subsequent field injection of bioaugmented groundwater, strain KTR9 (wild-type) migrated up to 23-m downgradient of the injection well within 3 days. Thus, the three RDX-degrading strains were effectively introduced and transported within the UMCD aquifer. This demonstration represents an innovative application of bioaugmentation to potentially enhance RDX biodegradation in aerobic aquifers.

Research paper thumbnail of Effect of Aging on the Biodegradation of Chemicals

Research paper thumbnail of Overview of in situ and ex situ bioremediation approaches for perchlorate

A mathematical model was developed to describe the biodegradation kinetics of perchlorate in the ... more A mathematical model was developed to describe the biodegradation kinetics of perchlorate in the presence of nitrate and oxygen as competing electron acceptors. The rate of perchlorate degradation is described as a function of the electron donor (acetate) degradation rate, the concentration of the alternate electron acceptors, and rates of biomass growth and decay. The kinetics of biomass growth are described using a modified Monod model, and inhibition factors are incorporated to describe the influence of oxygen and nitrate on perchlorate degradation. In order to develop input parameters for the model, a series of batch biodegradation studies were performed using Azospira suillum JPLRND, a perchlorate-degrading strain isolated from groundwater. This strain is capable of utilizing oxygen, nitrate, or perchlorate as terminal electron acceptors. The maximum specific growth rate (µ max ) and half-saturation constant (K don S ) for the bacterium when utilizing either perchlorate or nitrate were similar; 0.16 per h and 158 mg acetate/L, respectively. However, these parameters were different when the strain was growing on oxygen. In this case, µ max and K don S were 0.22 per h and 119 mg acetate/L, respectively. The batch experiments also revealed that nitrate inhibits perchlorate biodegradation by this strain. This finding was incorporated into the model by applying an inhibition coefficient (K nit i ) value of 25 mg nitrate/L. Combined with appropriate groundwater transport models, this model can be used to predict perchlorate biodegradation during in situ remediation efforts.

Research paper thumbnail of Using Stable Isotope Ratio Analysis to Distinguish Perchlorate Sources

Research paper thumbnail of ESTCP Project ER-200829

Research paper thumbnail of Mapping groundwater recharge areas for land use planning

Research paper thumbnail of Perchlorate Biodegradation for Water Treatment

Environmental Science & Technology, 2005

Research paper thumbnail of Rat kidney proximal tubule cells in defined medium: the roles of cholera toxin, extracellular calcium and serum in cell growth and expression of gamma-glutamyltransferase

In Vitro Cellular & Developmental Biology

A culture system is described in which rat kidney proximal tubule epithelial cells {RPTE) can be ... more A culture system is described in which rat kidney proximal tubule epithelial cells {RPTE) can be prepared with good yield and high viability and grown in culture under serum-free conditions. The cells require EGF, insulin, cholera toxin and either l% dialyzed serum or a complex of bovine serum albumin with oleic acid (BSA/OAk The cells can be maintained for long periods of time and express several markers for RPTE. The cells have both alkaline phosphatase and ?-glutamyltransferase activity and respond to parathyroid hormone but not vasopressin. The specific activity of y-glutamyltransferase decreases when the cells begin to grow, but increases when they reach confluence. Extracellular calcium plays a role in the induction of y-glutamyltransferase in confluent cells. Cells grown in media containing low calcium, i.e. less than 0.4 mM, have reduced specific activity of ),-glutamyltransferase. Extracellular calcium also alters the morphology of the cells in that cells grown in low calcium are single cells or loose clusters suggesting poor cell-cell contact. When the calcium is raised to 1.0 mM, the cells change their shape and organization to adopt the morphology of cells maintained continuously in 1.0 mM calcium. The cells can be passaged onto plastic surfaces which have been coated with collagen but cannot be subcultured on uncoated or serum coated plastic. This culture system will be a useful model for the investigation of renal carcinogenesis and the role of cell proliferation in that process. This paper reports a method for isolation and, uniquely, multipassage culture of rat proximal tubule epithelial cells. The authors use this culture system to explore some aspects of the physiology of these cells, demonstrating the value of the model.

Research paper thumbnail of Quantitation of multiple pathways for the metabolism of nephrotoxic cysteine conjugates using selective inhibitors of L-α-hydroxy acid oxidase (L-amino oxidase) and cysteine conjugate β-lyase

Drug Metabolism and Disposition

In this study, we have established the selectivity of inhibitors for rat kidney cysteine conjugat... more In this study, we have established the selectivity of inhibitors for rat kidney cysteine conjugate beta-lyase and L-alpha-hydroxy acid oxidase (L-amino acid oxidase) and have used these inhibitors to explore the relative roles of these two enzymes in the metabolism of nephrotoxic cysteine conjugates by rat kidney homogenate. In addition, we have investigated the relationship between structure and the metabolism of toxic cysteine conjugates by purified rat kidney L-alpha-hydroxy acid oxidase. With purified enzyme, S-(1,2,3,4,4-pentachlorobutadienyl)-L-cysteine (PCBDC) was about four times more active than S(1,2-dichlorovinyl)-L-cysteine (DCVC). Three alkyl conjugates were less active than DCVC. Purified L-alpha-hydroxy acid oxidase was not inhibited by the beta-lyase inhibitor aminooxyacetic acid but was inactivated by 2-hydroxy-3-butynoate. PCBDC metabolism in rat kidney homogenate was inhibited 74% by aminooxyacetic acid and 42% by 2-hydroxy-3-butynoate, whereas DCVC metabolism was inhibited 77% by aminooxyacetic acid and 28% by 2-hydroxy-3-butynoate. However, only aminooxyacetic acid inhibited the binding of 35S label from [35S]DCVC. Based on these results we have reached three conclusions. First, L-alpha-hydroxy acid oxidase plays a significant role in the metabolism of some cysteine conjugates. Second, metabolism of DCVC by L-alpha-hydroxy acid oxidase does not contribute directly to covalent binding. Third, as much as 65% of DCVC may be metabolized to its corresponding alpha-keto acid. The results are discussed with regard to the nephrotoxicity of cysteine conjugates.

Research paper thumbnail of Perchlorate and nitrate in situ bioremediation of ground water

Research paper thumbnail of Modeling In Situ Bioremediation of Perchlorate-Contaminated Groundwater

Perchlorate-contaminated groundwater is a significant national problem. An innovative technology ... more Perchlorate-contaminated groundwater is a significant national problem. An innovative technology was recently developed which uses a pair of dual-screened treatment wells to mix an electron donor into perchlorate- contaminated groundwater in order to effect in situ bioremediation of the perchlorate by indigenous perchlorate reducing bacteria (PRB) without the need to extract the contaminated water from the subsurface. The two treatment

Research paper thumbnail of In situ assessment of the transport and microbial consumption of oxygen in ground water, Cape Cod, Massachusetts

Research paper thumbnail of Guidance manual for forensic analysis of perchlorate in groundwater using chlorine and oxygen isotopic analyses

Research paper thumbnail of Field Demonstration of In Situ Perchlorate Bioremediation in Groundwater

Research paper thumbnail of Application of (13)C and (15)N stable isotope probing to characterize RDX degrading microbial communities under different electron-accepting conditions

Journal of hazardous materials, Jan 22, 2015

This study identified microorganisms capable of using the explosive hexahydro-1,3,5-trinitro-1,3,... more This study identified microorganisms capable of using the explosive hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) or its metabolites as carbon and/or nitrogen sources under different electron-accepting conditions using (13)C and (15)N stable isotope probing (SIP). Mesocosms were constructed using groundwater and aquifer solids from an RDX-contaminated aquifer. The mesocosms received succinate as a carbon source and one of four electron acceptors (nitrate, manganese(IV), iron(III), or sulfate) or no additional electron acceptor (to stimulate methanogenesis). When RDX degradation was observed, subsamples from each mesocosm were removed and amended with (13)C3- or ring-(15)N3-, nitro-(15)N3-, or fully-labeled (15)N6-RDX, followed by additional incubation and isolation of labeled nucleic acids. A total of fifteen 16S rRNA sequences, clustering in α- and γ-Proteobacteria, Clostridia, and Actinobacteria, were detected in the (13)C-DNA fractions. A total of twenty seven sequences were deri...

Research paper thumbnail of Factors affecting sequestration and bioavailability of phenanthrene in soils

Environmental Toxicology and Chemistry

A study was conducted to determine factors affecting the sequestration and changes in bioavailabi... more A study was conducted to determine factors affecting the sequestration and changes in bioavailability as phenanthrene persists in soils. Phenanthrene became sequestered in seven soils differing appreciably in organic matter and clay content as measured by earthworm uptake, bacterial mineralization, or extractability. Phenanthrene also became sequestered as it aged in soil aggregates of various sizes as measured by decline in availability to a bacterium, a mild extractant, or both. Wetting and drying a soil during aging reduced the amount of phenanthrene recovered by a mild extractant and the rate and extent of bacterial mineralization of the hydrocarbon. After biodegradation of phenanthrene added to the soil, more of the compound remained if it had been aged than if it had not been aged. Wetting and drying the soil during aging further increased the amount of phenanthrene remaining after biodegradation. The rate and extent of bacterial mineralization of phenanthrene were less in lea...

Research paper thumbnail of In-Situ and Ex-Situ Bioremediation Options for Treating Perchlorate in Groundwater

Remediation Journal, 2002

Research paper thumbnail of Active Bioremediation

SERDP/ESTCP Environmental Remediation Technology, 2009

Research paper thumbnail of Isotopic Tracing of Perchlorate in the Environment

Handbook of Environmental Isotope Geochemistry, 2011

Isotopic measurements can be used for tracing the sources and behavior of environmental contamina... more Isotopic measurements can be used for tracing the sources and behavior of environmental contaminants. Perchlorate (ClO 4 À ) has been detected

Research paper thumbnail of Applications of Selective Ion Exchange for Perchlorate Removal, Recovery and Environmental Forensics

Ion Exchange and Solvent Extraction Series, 2011

Isotopic measurements can be used for tracing the sources and behavior of environmental contamina... more Isotopic measurements can be used for tracing the sources and behavior of environmental contaminants. Perchlorate (ClO 4 À ) has been detected