Bulletin of Environmental Contamination and Toxicology, 2020
Polycyclic aromatic hydrocarbons (PAHs) and naphthenic acids (NAs) are toxic contaminants of envi... more Polycyclic aromatic hydrocarbons (PAHs) and naphthenic acids (NAs) are toxic contaminants of environmental concern found in process water and tailings from the oil sands industry. BioTiger™ (BT), a patented microbial consortium, was found to cometabolically biodegrade the NA hexanoic acid (HA) and the PAH phenanthrene (PH). BT also biodegrades both HA and PH when present in a mixture as well as biodegrading HA in the presence of phenanthrene and mature fine tailings (MFT). Hexanamide was produced and consumed during cometabolism of HA. Also, three of the BT strains were found to generate biosurfactants with the bacterial adhesion to hydrocarbons assay, seven with the MBAS assay, and nine with a hemolysis assay. Serial transfers of the BT consortium demonstrated the stability of HA degradation over several generations. The results demonstrate that BT cometabolically biodegrades various combinations of PH HA and that some components in the consortium produce biosurfactants in concert.
Rare earth elements (REEs) are crucial to many high-tech devices and maintaining a competitive mo... more Rare earth elements (REEs) are crucial to many high-tech devices and maintaining a competitive modern economy, thus they are of critical and strategic importance. Currently, mining approaches include rock grinding, strong acid or base extraction, separation, and concentration. The objective of this project was to develop cost-effective, environmentally friendly technologies for bioextracting REEs from mined materials.
Project Highlight Critical elements play a vital role in consumer electronics including computers... more Project Highlight Critical elements play a vital role in consumer electronics including computers, cell phones, medical equipment, as well as military hardware that are critical to the country's economic and national security. Strategies are needed to reduce our dependency on critical element imports. This project is developing approaches for biomining critical elements from geological materials. Project Team Robin L. Brigmon (PI) Charles Turick (retired 9/30/21), Daniel I. Kaplan, Anna Knox (co-PIs) Alex Kugler, a SRNL Post-doc starting his second year on the project.
BioTiger™ (BT) is a consortium of 12 bacteria designed for petroleum waste biodegradation. BT is ... more BioTiger™ (BT) is a consortium of 12 bacteria designed for petroleum waste biodegradation. BT is currently being studied and could be considered for bioremediation of the Athabasca oil sands refineries in Canada and elsewhere. The run off ponds from the petroleum extraction processes, called tailings ponds, are a mixture of polycyclic aromatic hydrocarbons, naphthenic acids, hydrocarbons, toxic chemicals like heavy metals, water, and sand. Due to environmental regulations the oil industry would like to separate and degrade the hazardous chemical species from the tailings ponds while recycling the water. It has been shown that BT at 30 C°is able to completely degrade 10 mM hexanoic acid (HA) cometabolically with 0.2% yeast extract (w/v) in 48 hours when starting at 0.4 OD 600nm. After establishing this stable degradation capability, variations were tested to explore the wider parameters of BT activity in temperature, pH, intermediate degradation, co-metabolic dependence, and transfer stability. Due to the vast differences in temperature at various points in the refineries, a wide range of temperatures were assessed. The results indicate that BT retains the ability to degrade HA, a model surrogate for tailings pond contaminants, at temperatures ranging from 15°C to 35°C. Hexanamide (HAM) was shown to be an intermediate generated during the degradation of HA in an earlier work and HAM is completely degraded after 48 hours, indicating that HAM is not the final product of HA degradation. Various replacements for yeast extract were attempted. Glucose, a carbon source; casein amino acids, a protein source; additional ammonia, mimicking known media; and additional phosphate with Wolffe's vitamins and minerals all showed no significant degradation of HA compared to control. Decreasing the yeast extract concentration (0.05%) demonstrated limited but significant degradation. Finally, serial inoculations of BT were performed to determine the stability of degradation over several generations. Overall, BT has shown to be moderately flexible for HA co-metabolic biodegradation.
This report was prepared as an account of work sponsored by an agency of the United States Govern... more This report was prepared as an account of work sponsored by an agency of the United States Government. Neither the United States Government nor any agency thereof, nor any of their employees, makes any warranty, express or implied, or assumes any legal liability or responsibility for the accuracy, completeness, or usefulness of any information, apparatus, product, or-process disclosed, or represents that its use would not infringe privately owned rights. Reference herein to any specific commercial product, process. or service by trade name, trademark, manufacturer, or otherwise does not necessarily constitute o r imply its endorsement. recommendation. or favoring by the United States Government or any agency thereof. The views and opinions of authors expressed herein do not necessarily state or reflect those of the United States-Government or any agency thereof. This report has been reproduced directly from the best available copy. Available to DOE and DOE contractors from the Office of Scientific and Technical Information, P. 0. Box 62, Oak Ridge, TN 37831: prices available from (615) 576-8401.
increased in treated soil about 16-fold, 10-fold and 2-fold, respectively. Changes in the biodive... more increased in treated soil about 16-fold, 10-fold and 2-fold, respectively. Changes in the biodiversity of bacterial populations were evaluated by the Ecophysiological Index (EP) and the Colony Development Index (CD) for oligotrophs and copiotrophs. During the experimental period slower growing microorganisms (K-strategists) predominated. The application of amendments to the soil led to an increase of the CD index in both copiotroph and oligotroph populations after 28 weeks. EP, CD, bacteria, actinomycetes and fungi increased in the treated soil. Traditional microbiological methods based on culture techniques can be used to evaluate the biological quality of the phytostabilized heavy metal-contaminated soil.
Comparative Biochemistry and Physiology Part A: Physiology, 1994
/I-Hydroxybutyrate and routine biochemical parameters were determined on plasma and urine samples... more /I-Hydroxybutyrate and routine biochemical parameters were determined on plasma and urine samples obtained from free-ranging desert tortoises over a l-year period. Urea nitrogen varied inversely with total protein concentration (P < 0.019); significant azotemia and hypoproteinemia were noted following emergence from hibernation. In summer, elevated glucose and cholesterol levels, together with lower sodium, urea nitrogen and urine specific gravity, reflected availability of food and water. Plasma /l-hydroxybutyrate levels signiilcantly (P < 0.004) increased in the fall (1.92 _+ 0.95 mmol/l), during a period of low rainfall. Results suggest that plasma j?-hydroxybutyrate may increase in response to short term energy needs, while protein catabolism plays a greater role in providing fuel during hibernation.
Adult male and female Beagle dogs (eight total) were exposed individually, in series, to each of ... more Adult male and female Beagle dogs (eight total) were exposed individually, in series, to each of 23 effective temperatures for a period of 2 hours or until rectal temperature increased 1.1 degrees C. Rectal temperature was measured to the nearest 0.1 degree C by thermistor probes in the pre-test condition (basal temperature) and at each 5-minute interval during the test conditions (effective temperatures between 21.1 degrees C and 34.7 degrees C). The frequency at which dogs displayed a 1.1 degree C rise in rectal temperature was related to the magnitude of the effective temperature. At an effective temperature of 32.6 degrees C or greater, 100% of the dogs displayed a 1.1 degree C rise in rectal temperature. Between an effective temperature of 29.3 degrees C and 31.4 degrees C, some animals displayed a 1.1 degree C rise while others did not. At an effective temperature of 28.4 degrees C or below no animals displayed a 1.1 degree C rise. The mean time necessary for a 1.1 degree C ri...
An inexpensive but reliable telemetry system for long-term, sequential monitoring of body tempera... more An inexpensive but reliable telemetry system for long-term, sequential monitoring of body temperature in up to 20 laboratory animals is described. The system consists of frequency-modulated (FM) temperature transmitters, remote-controlled power switches to extend battery life, a multi-channel telemetry receiver, and a frequency counter interfaced with a personal computer to record data. Analysis of body temperature data obtained from four New Zealand White rabbits confirms the reliability and value of this system.
ABSTRACT Due to its chemical stability, uniform pore size, and high pore density, nanoporous alum... more ABSTRACT Due to its chemical stability, uniform pore size, and high pore density, nanoporous alumina is being investigated for use in biosensing, drug delivery, hemodialysis, and other medical applications. In recent work, we have examined the use of atomic layer deposition for coating the surfaces of nanoporous alumina membranes. Zinc oxide coatings were deposited on nanoporous alumina membranes using atomic layer deposition. The zinc oxide-coated nanoporous alumina membranes demonstrated antimicrobial activity against Escherichia coli and Staphylococcus aureus bacteria. These results suggest that atomic layer deposition is an attractive technique for modifying the surfaces of nanoporous alumina membranes and other nanostructured biomaterials.
World Journal of Microbiology and Biotechnology, 2006
Fate of benzene ethylbenzene toluene xylenes (BTEX) compounds through biodegradation was investig... more Fate of benzene ethylbenzene toluene xylenes (BTEX) compounds through biodegradation was investigated using two different bacteria, Ralstonia picketti (BP-20) and Alcaligenes piechaudii (CZOR L-1B). These bacteria were isolated from extremely polluted petroleum hydrocarbon contaminated soils. PCR and Fatty Acid Methyl Ester (FAME) were used to identify the isolates. Biodegradation was measured using each organism individually and in combination. Both bacteria were shown to degrade each of the BTEX compounds. Alcaligenes piechaudii biodegraded BTEXs more efficiently while mixed with BP-20 and individually. Biosurfactant production was observed by culture techniques. In addition 3hydroxy fatty acids, important in biosurfactant production, was observed by FAME analysis. In the all experiments toluene and m+p-xylenes were better growth substrates for both bacteria than the other BTEX compounds. In addition, the test results indicate that the bacteria could contribute to bioremediation of aromatic hydrocarbons (BTEX) pollution increase biodegradation through the action by biosurfactants.
Description/Abstract The degradation of petroleum waste by mixed bacterial cultures which produce... more Description/Abstract The degradation of petroleum waste by mixed bacterial cultures which produce biosurfactants: Ralstonia pickettii SRS (BP-20), Alcaligenes piechaudii SRS (CZOR L-1B), Bacillus subtilis (1'-1a), Bacillus sp.(T-1) and Bacillus sp.(T'-1) was investigated. ...
Immunological procedures were developed to enumerate chlorinated hydrocarbon (CH) degrading bacte... more Immunological procedures were developed to enumerate chlorinated hydrocarbon (CH) degrading bacteria in environmental samples. Polyclonal antibodies (Pabs) were produced by immunizing New Zealand white rabbits against 18 CH-degrading bacteria. These bacteria included methanotrophic and chlorobenzene (CB)-degrading species. An enzyme-linked immunosorbent assay (ELISA) was used to test for cross-reactivity and sensitivity of the Pabs. Direct fluorescent antibodies (DFAs) were developed against selected methanotrophic bacteria isolated from a trichloroethylene (TCE)-contaminated landfill at the Savannah River Site (SRS) and cultures from the American Type Culture Collection (ATCC). Analysis of ELISA results showed some of the Pabs to be group specific while others were species specific. The threshold of sensitivity for the ELISA 5 is 10 bacteria cells / ml. Results from the DFA and ELISA techniques for enumeration of methanotrophic bacteria in groundwater were higher but not significantly different (P,0.05) compared to indirect microbiological techniques such as MPN, although the direct enumerations were accomplished in significantly less time. These methods provide important information on in situ microbial community structure and function for bioremediation applications within 1-4 h of sampling.
Bulletin of Environmental Contamination and Toxicology, 2020
Polycyclic aromatic hydrocarbons (PAHs) and naphthenic acids (NAs) are toxic contaminants of envi... more Polycyclic aromatic hydrocarbons (PAHs) and naphthenic acids (NAs) are toxic contaminants of environmental concern found in process water and tailings from the oil sands industry. BioTiger™ (BT), a patented microbial consortium, was found to cometabolically biodegrade the NA hexanoic acid (HA) and the PAH phenanthrene (PH). BT also biodegrades both HA and PH when present in a mixture as well as biodegrading HA in the presence of phenanthrene and mature fine tailings (MFT). Hexanamide was produced and consumed during cometabolism of HA. Also, three of the BT strains were found to generate biosurfactants with the bacterial adhesion to hydrocarbons assay, seven with the MBAS assay, and nine with a hemolysis assay. Serial transfers of the BT consortium demonstrated the stability of HA degradation over several generations. The results demonstrate that BT cometabolically biodegrades various combinations of PH HA and that some components in the consortium produce biosurfactants in concert.
Rare earth elements (REEs) are crucial to many high-tech devices and maintaining a competitive mo... more Rare earth elements (REEs) are crucial to many high-tech devices and maintaining a competitive modern economy, thus they are of critical and strategic importance. Currently, mining approaches include rock grinding, strong acid or base extraction, separation, and concentration. The objective of this project was to develop cost-effective, environmentally friendly technologies for bioextracting REEs from mined materials.
Project Highlight Critical elements play a vital role in consumer electronics including computers... more Project Highlight Critical elements play a vital role in consumer electronics including computers, cell phones, medical equipment, as well as military hardware that are critical to the country's economic and national security. Strategies are needed to reduce our dependency on critical element imports. This project is developing approaches for biomining critical elements from geological materials. Project Team Robin L. Brigmon (PI) Charles Turick (retired 9/30/21), Daniel I. Kaplan, Anna Knox (co-PIs) Alex Kugler, a SRNL Post-doc starting his second year on the project.
BioTiger™ (BT) is a consortium of 12 bacteria designed for petroleum waste biodegradation. BT is ... more BioTiger™ (BT) is a consortium of 12 bacteria designed for petroleum waste biodegradation. BT is currently being studied and could be considered for bioremediation of the Athabasca oil sands refineries in Canada and elsewhere. The run off ponds from the petroleum extraction processes, called tailings ponds, are a mixture of polycyclic aromatic hydrocarbons, naphthenic acids, hydrocarbons, toxic chemicals like heavy metals, water, and sand. Due to environmental regulations the oil industry would like to separate and degrade the hazardous chemical species from the tailings ponds while recycling the water. It has been shown that BT at 30 C°is able to completely degrade 10 mM hexanoic acid (HA) cometabolically with 0.2% yeast extract (w/v) in 48 hours when starting at 0.4 OD 600nm. After establishing this stable degradation capability, variations were tested to explore the wider parameters of BT activity in temperature, pH, intermediate degradation, co-metabolic dependence, and transfer stability. Due to the vast differences in temperature at various points in the refineries, a wide range of temperatures were assessed. The results indicate that BT retains the ability to degrade HA, a model surrogate for tailings pond contaminants, at temperatures ranging from 15°C to 35°C. Hexanamide (HAM) was shown to be an intermediate generated during the degradation of HA in an earlier work and HAM is completely degraded after 48 hours, indicating that HAM is not the final product of HA degradation. Various replacements for yeast extract were attempted. Glucose, a carbon source; casein amino acids, a protein source; additional ammonia, mimicking known media; and additional phosphate with Wolffe's vitamins and minerals all showed no significant degradation of HA compared to control. Decreasing the yeast extract concentration (0.05%) demonstrated limited but significant degradation. Finally, serial inoculations of BT were performed to determine the stability of degradation over several generations. Overall, BT has shown to be moderately flexible for HA co-metabolic biodegradation.
This report was prepared as an account of work sponsored by an agency of the United States Govern... more This report was prepared as an account of work sponsored by an agency of the United States Government. Neither the United States Government nor any agency thereof, nor any of their employees, makes any warranty, express or implied, or assumes any legal liability or responsibility for the accuracy, completeness, or usefulness of any information, apparatus, product, or-process disclosed, or represents that its use would not infringe privately owned rights. Reference herein to any specific commercial product, process. or service by trade name, trademark, manufacturer, or otherwise does not necessarily constitute o r imply its endorsement. recommendation. or favoring by the United States Government or any agency thereof. The views and opinions of authors expressed herein do not necessarily state or reflect those of the United States-Government or any agency thereof. This report has been reproduced directly from the best available copy. Available to DOE and DOE contractors from the Office of Scientific and Technical Information, P. 0. Box 62, Oak Ridge, TN 37831: prices available from (615) 576-8401.
increased in treated soil about 16-fold, 10-fold and 2-fold, respectively. Changes in the biodive... more increased in treated soil about 16-fold, 10-fold and 2-fold, respectively. Changes in the biodiversity of bacterial populations were evaluated by the Ecophysiological Index (EP) and the Colony Development Index (CD) for oligotrophs and copiotrophs. During the experimental period slower growing microorganisms (K-strategists) predominated. The application of amendments to the soil led to an increase of the CD index in both copiotroph and oligotroph populations after 28 weeks. EP, CD, bacteria, actinomycetes and fungi increased in the treated soil. Traditional microbiological methods based on culture techniques can be used to evaluate the biological quality of the phytostabilized heavy metal-contaminated soil.
Comparative Biochemistry and Physiology Part A: Physiology, 1994
/I-Hydroxybutyrate and routine biochemical parameters were determined on plasma and urine samples... more /I-Hydroxybutyrate and routine biochemical parameters were determined on plasma and urine samples obtained from free-ranging desert tortoises over a l-year period. Urea nitrogen varied inversely with total protein concentration (P < 0.019); significant azotemia and hypoproteinemia were noted following emergence from hibernation. In summer, elevated glucose and cholesterol levels, together with lower sodium, urea nitrogen and urine specific gravity, reflected availability of food and water. Plasma /l-hydroxybutyrate levels signiilcantly (P < 0.004) increased in the fall (1.92 _+ 0.95 mmol/l), during a period of low rainfall. Results suggest that plasma j?-hydroxybutyrate may increase in response to short term energy needs, while protein catabolism plays a greater role in providing fuel during hibernation.
Adult male and female Beagle dogs (eight total) were exposed individually, in series, to each of ... more Adult male and female Beagle dogs (eight total) were exposed individually, in series, to each of 23 effective temperatures for a period of 2 hours or until rectal temperature increased 1.1 degrees C. Rectal temperature was measured to the nearest 0.1 degree C by thermistor probes in the pre-test condition (basal temperature) and at each 5-minute interval during the test conditions (effective temperatures between 21.1 degrees C and 34.7 degrees C). The frequency at which dogs displayed a 1.1 degree C rise in rectal temperature was related to the magnitude of the effective temperature. At an effective temperature of 32.6 degrees C or greater, 100% of the dogs displayed a 1.1 degree C rise in rectal temperature. Between an effective temperature of 29.3 degrees C and 31.4 degrees C, some animals displayed a 1.1 degree C rise while others did not. At an effective temperature of 28.4 degrees C or below no animals displayed a 1.1 degree C rise. The mean time necessary for a 1.1 degree C ri...
An inexpensive but reliable telemetry system for long-term, sequential monitoring of body tempera... more An inexpensive but reliable telemetry system for long-term, sequential monitoring of body temperature in up to 20 laboratory animals is described. The system consists of frequency-modulated (FM) temperature transmitters, remote-controlled power switches to extend battery life, a multi-channel telemetry receiver, and a frequency counter interfaced with a personal computer to record data. Analysis of body temperature data obtained from four New Zealand White rabbits confirms the reliability and value of this system.
ABSTRACT Due to its chemical stability, uniform pore size, and high pore density, nanoporous alum... more ABSTRACT Due to its chemical stability, uniform pore size, and high pore density, nanoporous alumina is being investigated for use in biosensing, drug delivery, hemodialysis, and other medical applications. In recent work, we have examined the use of atomic layer deposition for coating the surfaces of nanoporous alumina membranes. Zinc oxide coatings were deposited on nanoporous alumina membranes using atomic layer deposition. The zinc oxide-coated nanoporous alumina membranes demonstrated antimicrobial activity against Escherichia coli and Staphylococcus aureus bacteria. These results suggest that atomic layer deposition is an attractive technique for modifying the surfaces of nanoporous alumina membranes and other nanostructured biomaterials.
World Journal of Microbiology and Biotechnology, 2006
Fate of benzene ethylbenzene toluene xylenes (BTEX) compounds through biodegradation was investig... more Fate of benzene ethylbenzene toluene xylenes (BTEX) compounds through biodegradation was investigated using two different bacteria, Ralstonia picketti (BP-20) and Alcaligenes piechaudii (CZOR L-1B). These bacteria were isolated from extremely polluted petroleum hydrocarbon contaminated soils. PCR and Fatty Acid Methyl Ester (FAME) were used to identify the isolates. Biodegradation was measured using each organism individually and in combination. Both bacteria were shown to degrade each of the BTEX compounds. Alcaligenes piechaudii biodegraded BTEXs more efficiently while mixed with BP-20 and individually. Biosurfactant production was observed by culture techniques. In addition 3hydroxy fatty acids, important in biosurfactant production, was observed by FAME analysis. In the all experiments toluene and m+p-xylenes were better growth substrates for both bacteria than the other BTEX compounds. In addition, the test results indicate that the bacteria could contribute to bioremediation of aromatic hydrocarbons (BTEX) pollution increase biodegradation through the action by biosurfactants.
Description/Abstract The degradation of petroleum waste by mixed bacterial cultures which produce... more Description/Abstract The degradation of petroleum waste by mixed bacterial cultures which produce biosurfactants: Ralstonia pickettii SRS (BP-20), Alcaligenes piechaudii SRS (CZOR L-1B), Bacillus subtilis (1'-1a), Bacillus sp.(T-1) and Bacillus sp.(T'-1) was investigated. ...
Immunological procedures were developed to enumerate chlorinated hydrocarbon (CH) degrading bacte... more Immunological procedures were developed to enumerate chlorinated hydrocarbon (CH) degrading bacteria in environmental samples. Polyclonal antibodies (Pabs) were produced by immunizing New Zealand white rabbits against 18 CH-degrading bacteria. These bacteria included methanotrophic and chlorobenzene (CB)-degrading species. An enzyme-linked immunosorbent assay (ELISA) was used to test for cross-reactivity and sensitivity of the Pabs. Direct fluorescent antibodies (DFAs) were developed against selected methanotrophic bacteria isolated from a trichloroethylene (TCE)-contaminated landfill at the Savannah River Site (SRS) and cultures from the American Type Culture Collection (ATCC). Analysis of ELISA results showed some of the Pabs to be group specific while others were species specific. The threshold of sensitivity for the ELISA 5 is 10 bacteria cells / ml. Results from the DFA and ELISA techniques for enumeration of methanotrophic bacteria in groundwater were higher but not significantly different (P,0.05) compared to indirect microbiological techniques such as MPN, although the direct enumerations were accomplished in significantly less time. These methods provide important information on in situ microbial community structure and function for bioremediation applications within 1-4 h of sampling.
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