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R J Larson

Publications and source records attributed to R J Larson.

At least 19 recordsLinked to original sources
PubMed2001

Structure of microbial communities in activated sludge: potential implications for assessing the biodegradability of chemicals.

Various methods used to assess the biodegradability of chemicals often employ activated sludge as an inoculum since chemicals that ultimately enter the environment are often discharged through wastewater. Differences in the structure and function of activated sludge microbial communities that may complicate interpretation of biodegradation tests could arise from differences in wastewater composition, wastewater treatment plant (WWTP) operation, or manipulations done after collection of the activated sludge. In this study, various methods were used to characterize the structure of microbial communities found in freshly collected activated sludge from WWTPs in Japan, Europe, and the United States, as well as sludge that had been continuously fed either sewage or a glucose-peptone mixture for several weeks after collection. Comparisons of biomass levels, whole-community substrate utilization (determined using Biolog GN and GP plates), and phospholipid fatty acid (PLFA) profiles indicated there were both geographical and temporal differences among freshly collected activated sludge samples. Moreover, marked shifts in the structure of activated sludge microbial communities occurred upon continuous cultivation in the laboratory for 5 weeks using a glucose-peptone feed. These shifts were evident from whole-community substrate utilization and PLFA profiles as well as differences in the profiles of 16S rDNA genes from numerically dominant populations obtained by denaturing gradient gel electrophoresis and terminal restriction fragment analyses. Further studies are needed to better define the variability within and between activated sludge from wastewater treatment plants and laboratory reactors and to assess the impact of such differences on the outcome of biodegradability tests.

Bacteria↗
PubMed2001

Characterization and distribution of esterase activity in activated sludge.

The location and activity of esterase enzymes in activated sludge from three municipal wastewater treatment plants were characterized using model substrates and denaturing and non-denaturing polyacrylamide gel electrophoresis (PAGE) of particulate, freeze-thaw (primarily periplasmic enzymes and those associated with outer cell surfaces) and extracellular fractions of activated sludge bacteria. Particulate and freeze-thaw fractions had a similar spectrum of substrate specificity and contained significant levels of protein and esterase activity against model substrates, C2-C18 monoesters of p-nitrophenol and C2-C8 diesters of fluorescein. Esterase activity was highest with substrates that had short alkyl chains (C4) and decreased as the chain lengths increased beyond C8. Extracellular fractions contained very low levels of protein (<0.1 mg/l) and showed no esterase activity against any of the model substrates tested. Multiple bands were observed upon analysis of particulate and freeze-thaw fractions by non-denaturing PAGE in combination with activity staining using various alpha-naphthol ester substrates (C2-C8). Our results indicate that esterase enzymes in activated sludge are fairly diverse from a structural standpoint but exhibit a high level of functional redundancy, with different enzymes catalyzing the same reactions in different sludges. Extracellular esterase activity was totally absent for the substrates we tested and the esterase activity that we observed was closely linked to a particulate floc or cellular material.

Electrophoresis, Polyacrylamide Gel↗
PubMed2000

Evaluation of an inactivation procedure for determining the sorption of organic compounds to activated sludge.

A batch method was developed and validated for determining the sorption coefficient (Kd) of biodegradable organic compounds to activated sludge solids using a nonspecific analytical technique, total organic carbon (TOC) analysis. In this method, activated sludge solids were first inactivated by lyophilization and dry heat (103 degrees C) and then washed to remove any dissolved organic carbon released during the inactivation procedure. The inactivated sludge solids were exposed to a range of concentrations of different test compounds in synthetic wastewater until equilibrium was achieved (< 2 h). The amount of test compound sorbed to the solids was then determined by measuring TOC levels in centrifuged supernatants. Results revealed that the sorption coefficients (Kd values) for four detergent chemicals in inactivated sludge solids using TOC analysis were in good agreement with values determined in fresh activated sludge using radiolabeled materials. These Kd values are suitable for use in estimating environmental exposure concentrations and for developing screening-level models to assess the removal of organic compounds by sorption and settling during activated sludge wastewater treatment.

Adsorption↗
PubMed1998

Chicken ovalbumin upstream promoter-transcription factor (COUP-TF) modulates expression of the Purkinje cell protein-2 gene. A potential role for COUP-TF in repressing premature thyroid hormone action in the developing brain.

The cerebellar Purkinje cell-specific PCP-2 gene is transcriptionally activated by thyroid hormone during the 2nd and 3rd weeks of postnatal life in the rat. In contrast, thyroid hormone has no detectable effects on PCP-2 expression in the fetal rat. We now present data that suggest that the orphan nuclear receptor chicken ovalbumin upstream promoter-transcription factor (COUP-TF) represses triiodothyronine (T3)-dependent transcriptional activation of PCP-2 in the immature Purkinje cell. Gel shift assays show that the PCP-2 A1TRE and adjoining sequences (-295/-199 region) bind to rat and mouse brain nucleoproteins in a developmentally regulated fashion and that one of these nucleoproteins could be the orphan nucleoprotein COUP-TF. In support of this hypothesis, in vitro translated COUP-TF binds to the -295/-199 region and COUP-TF represses T3-dependent activation of the PCP-2 promoter in transient transfection analyses. Finally, immunohistochemical studies reveal that COUP-TF is specifically expressed in the immature fetal and early neonatal Purkinje cell and that this expression diminishes coincident with thyroid hormone induction of PCP-2 expression. Our findings are consistent with the hypothesis that the presence or absence of inhibitory proteins bound to the thyroid hormone response element of T3-responsive genes governs the responsivity of these genes to thyroid hormone during brain development.

Animals↗
PubMed1998

A simplified modeling approach using microbial growth kinetics for predicting exposure concentrations of organic chemicals in treated wastewater effluents.

Various mathematical relationships have been used to assess exposure concentrations of organic chemicals when emissions occur via wastewater treatment. These relationships range from a simple removal factor calculation to more sophisticated approaches using kinetic based mathematical models. While these existing approaches have been used by decision makers to screen new chemicals for exposure assessments, they all have limitations in the predictive capabilities. Thus, a simplified modeling approach grounded in sound scientific fundamentals that utilizes relatively easy to obtain input parameters is needed. In this paper a simplified modeling approach that utilizes microbial growth kinetics was developed for predicting effluent concentrations in secondary biological wastewater treatment systems. Receiving water predicted exposure concentrations (PEC) are assessed by using a dilution factor. One advantage of this approach is that it allows for wastewater treatment plant effluent concentrations, and therefore receiving water exposure levels, to be predicted with a minimum amount of experimental data. It also provides quantitative data that can be used to assess the relative biodegradability of different chemicals for use in regulatory and risk assessment activities.

Bacteria↗
PubMed1997

Purkinje cell protein-2 cis-elements mediate repression of T3-dependent transcriptional activation.

Previous studies in our laboratory show that triiodothyronine upregulates expression of the cerebellar Purkinje cell-specific gene Pcp-2 during the first 2 weeks of rat neonatal life. A specific thyroid hormone response element, the A1 TRE, mediates this regulation. The finding that the contiguous 68 bases (-267/ -199) of the Pcp-2 promoter 3' to the A1 TRE repressed T3 response in transactivation studies suggested that this sequence could play a role in preventing premature T3-dependent activation of Pcp-2 in the fetus. We now show that deletion of this region resulted in enhanced T3-dependent activation of the native Pcp-2 promoter. The sequence is not a generalized silencer since it does not alter basal activity of mouse mammary tumor virus (MMTV) or thymidine kinase (TK) promoters. Deletion and linker scanning studies indicate that the 5' 30 bases of the -267/ -199 region mediate most of the response silencing activity. The -267/ -199 region also attenuates T3-induced transactivation mediated by other TREs. Gel shift analysis reveals that nuclear proteins from fetal but not adult brains complex with the -267/ -199 region, supporting the hypothesis that this region binds proteins that suppress Pcp-2 expression early in brain development.

Animals↗
PubMed1997

Biodegradation of [S,S], [R,R] and mixed stereoisomers of ethylene diamine disuccinic acid (EDDS), a transition metal chelator.

An in-depth biodegradation test program was executed on the hexadentate ligand Ethylene Diamine Di Succinate (EDDS). The EDDS structure contains two chiral carbon atoms, and has three stereoisomers ([R,R], [R,S]/[S,R], [S,S]). Our research has focused on the isomer mixture (i.e. 25%[S,S]; 25%[R,R]; 50%[S,R]/[R,S], as produced from the reaction of ethylene diamine with maleic anhydride) and on the single [S,S]- and [R,R]-isomers. Biodegradation screening of the 14C-labelled EDDS isomer mixture in a Batch Activated Sludge (BAS) test with various inocula revealed incomplete mineralization, up to ca. 65% after 28 days. N-(2-aminoethyl) aspartic acid (AEAA), probably the d-isomer, was identified as the major portion of the 14C-material remaining in solution. Further testing revealed that the [S,S]-isomer is rapidly and completely mineralized in all test systems. By contrast, [R,R]-EDDS remained undegraded in a Sturm (OECD 301B) test, but was very slowly biotransformed into the recalcitrant metabolite AEAA in a BAS test. The [S,R]/[R,S] form undergoes biotransformation to AEAA in both high and low biomass systems. In a sewage treatment simulation test (OECD 303) the steady state DOC removal of mixture-EDDS in a CAS test was limited to 25-35%, even after extensive pre-acclimation, while the [S,S]-isomer achieved nearly complete removal (96%). This study illustrates the importance stereospecificity may have on the biodegradation and metabolite formation of a chemical. A biodegradation scheme for the different EDDS stereoisomers is proposed.

Aspartic Acid↗
PubMed1996

Estimating the removal and biodegradation potential of radiolabeled organic chemicals in activated sludge.

A two-step procedure is described to characterize the removal and biodegradation potential of nonvolatile 14C-labeled organic compounds in activated sludge. In the first step, trace concentrations of radiolabeled test materials are dosed in influent wastewater to continuous-flow activated sludge (CAS) systems which have been previously exposed or acclimated to unlabeled test material. Radiolabel is quantified in influent, effluent, and activated sludge mixed liquor to determine total 14C removal and partitioning of radiolabel in solid and liquid compartments. The 14C data are used to calculate the amount of removal due to sorption and biodegradation and to estimate the apparent sorption coefficients for 14C activity to activated sludge solids. The 14C-labeled CAS studies are followed by biodegradation studies in batch-activated sludge (BAS) systems using sludge derived from the CAS system. The kinetics of biodegradation (defined as mineralization to 14CO2) are measured in the BAS system to confirm the CAS biodegradation results and generate mineralization rate constants for kinetic modeling. The two-step procedure was applied to radiolabeled anionic (linear alkylbenzene sulfonate) and cationic (dodecyltrimethylammonium chloride, distearyldimethylammonium chloride) surfactants which differed greatly in their biodegradation and sorption profiles. Laboratory removal figures for these materials were comparable to values measured in full-scale wastewater treatment systems, although the amount of removal due to sorption and biodegradation varied significantly for the different surfactants. In general, the 14C method has several advantages over standard methods used in the United States and Europe which employ unlabeled materials. These advantages include the use of realistic concentrations and test conditions for acclimating and dosing activated sludge microorganisms and the ability to generate partitioning and kinetic constants that can be used more broadly in environmental fate and exposure models.

Biodegradation, Environmental↗
PubMed1996

A Purkinje cell protein-2 intronic thyroid hormone response element binds developmentally regulated thyroid hormone receptor-nuclear protein complexes.

Two thyroid hormone response elements (TREs), designated A1 TRE (-295/-268) and B1 TRE (+207/+227), have been identified within the Purkinje cell-expressed Pcp-2 gene. Previous studies have characterized the A1 TRE (Zou et al., 1994). This article analyzes the structural and functional characteristics of the intronic B1 TRE. The B1 sequence contains four overlapping TRE half-sites. The 3' DR4 motif, consisting of the second and forth half-sites, is responsible for the T3 induction observed with the B1 sequence. Gel-shift analysis reveals developmentally regulated complexes that are abundant in the fetus and at birth and then fall precipitously in the neonate bind to B1. The observed time-course of these complexes varies inversely with the rise in Pcp-2 expression, thus raising the possibility that the complexes may represent inhibitory factors. Supershift analysis indicates that endogenous TR alpha 1 is present in the fetal nuclear protein complexes that bind to B1. Competition analysis also indicates the second B1 TRE half-site is important in binding the TR alpha 1-TRAP complexes. These studies suggest that the B1 sequence may bind potential TR alpha 1-TRAP repressor complexes in the fetus, whereas in the neonate, these TRE sites may be involved in the activation of Pcp-2 by binding other TR-TRAP-activating complexes.

Animals↗
PubMed1996

Carbon dioxide recovery in ready biodegradation tests: mass transfer and kinetic considerations.

The kinetics and efficiency of carbon dioxide recovery in modern versions of the Sturm Ready Biodegradation Test were examined to determine the ability of CO2 evolution measurements to accurately estimate the rate and extent of ultimate biodegradation (mineralization). Kinetic data were analyzed by nonlinear regression techniques using an automated curve-fitting package available from commercial sources. The kinetics of CO2 recovery in standard 3.8 L glass carboys containing 2 L of medium were rapid when headspace aeration (approximately 6 ml/min) and moderate agitation (140 rev/min) on a rotary platform shaker were used to ensure adequate aeration and mixing. The time (half-life) for 50% CO2 recovery in external base traps was 4-5 hours, and stoichiometric recoveries of CO2 equivalents added as bicarbonate were obtained within 24 hours. The kinetics of CO2 evolution during biodegradation of several test compounds were significantly slower than the kinetics of CO2 recovery, with half-lives between 65 and 191 hours. Our results indicate that mass transfer limitations do not impact CO2 recoveries or biodegradation kinetic measurements in modern versions of the Sturm Test, even in test vessels with relatively low surface area to volume ratios (1:1). The use of headspace aeration and mixing generates reliable kinetic data, which can be analyzed by commercially-available nonlinear regression packages to provide rate information for the classification of chemicals with different biodegradation profiles.

Biodegradation, Environmental↗
PubMed1996

Impact of biodegradation test methods on the development and applicability of biodegradation QSARs.

The biodegradability of a substance depends on the structure and physical form of the substance, the time that has been available for acclimation, and the environmental conditions. Importantly, these later factors can be just as important as structure in determining the outcome of a biodegradation test. The development of appropriate QSARs for biodegradation and the ultimate value of the final QSAR depends on understanding these factors. This paper will describe what is known about the effect of test conditions on the results of biodegradation tests. The ability of these tests to reflect real environmental conditions will also be examined. Finally, we will discuss what we believe, in the light of this information, should be the goal of biodegradation QSARs and how these QSARs can be most appropriately used in fate assessments.

Bacteria↗
PubMed1995

Integrated approach for environmental assessment of new and existing substances.

To ensure the environmental safety of new and existing substances, the environmental fate and potential effects resulting from their release into the environment must be assessed. This requires the development of reasonable, consistent, and effective methods to conduct environmental risk assessments and to prioritize testing for these substances. This assessment must integrate fate and effects at the point-of-entry; it should also extend to an assessment of the potential to persist, and the consequences of increases in exposure concentrations, and to bioaccumulate. The conventional environmental risk assessment approach is used to assess the fate and effects of a substance at its point-of-entry into the environment. In this paper, an approach is presented for conducting quantitative environmental risk assessments of new and existing substances that builds on this conventional approach by including quantitative assessment of the potential for, and consequences of, persistence and bioaccumulation. The approach is described for aquatic, sediment, and terrestrial environments. For each environmental compartment, the approach includes (i) classification of the substance, based on environmental partitioning processes, to ensure that the appropriate data are collected and models used; (ii) a fate assessment to understand the ultimate fate of the substance after entry into the environment or "an environmental compartment" and to predict the exposure concentration of the substance at point-of-entry; (iii) a persistence assessment which determines the potential for increase in the exposure concentration as a result of repeated additions of the substance; (iv) effects assessment; (v) environmental risk assessment to examine the potential for adverse impact on ecosystems; and (vi) a bioaccumulation assessment to evaluate the potential for direct and indirect effects on the species of interest due to bioaccumulation. The assessment approach is illustrated using data for a hypothetical consumer product substance that is disposed down-the-drain.

Animals↗
PubMed1992

Integrated approach to surfactant environmental safety assessment: fast atom bombardment mass spectrometry and liquid scintillation counting to determine the mechanism and kinetics of surfactant biodegradation.

Fast atom bombardment mass spectrometry and liquid scintillation counting have been used to study the biodegradation of a novel cationic surfactant in live sludge. The rates of primary biodegradation and the extent of complete mineralization were determined. Furthermore, an intermediate degradation product was identified and its rates of formation and subsequent removal have been established. These data find utility in assessing the environmental safety of the surfactant and the accuracy of various environmental fate models.

Biodegradation, Environmental↗
PubMed1992

Aspartate aminotransferase in effective and ineffective alfalfa nodules : cloning of a cDNA and determination of enzyme activity, protein, and mRNA levels.

Aspartate aminotransferase (AAT) is a key plant enzyme affecting nitrogen and carbon metabolism, particularly in legume root nodules and leaves of C(4) species. To ascertain the molecular genetic characteristics and biochemical regulation of AAT, we have isolated a cDNA encoding the nodule-enhanced AAT (AAT-2) of alfalfa (Medicago sativa L.) by screening a root nodule cDNA expression library with antibodies. Complementation of an Escherichia coli AAT mutant with the alfalfa nodule AAT-2 cDNA verified the identity of the clone. The deduced amino acid sequence of alfalfa AAT-2 is 53 and 47% identical to animal mitochondrial and cytosolic AATs, respectively. The deduced molecular mass of AAT-2 is 50,959 daltons, whereas the mass of purified AAT-2 is about 40 kilodaltons as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and the protein's N-terminal domain (amino acids 1-59) contains many of the characteristics of plastid-targeting peptides. We postulate that AAT-2 is localized to the plastid. Southern blot analysis suggests that AAT-2 is encoded by a small, multigene family. The expression of AAT-2 mRNA in nodules is severalfold greater than that in either leaves or roots. Northern and western blots showed that expression of AAT activity during effective nodule development is accompanied by a sevenfold increase in AAT-2 mRNA and a comparable increase in enzyme protein. By contrast, plant-controlled ineffective nodules express AAT-2 mRNA at much lower levels and have little to no AAT-2 enzyme protein. Expression of root nodule AAT-2 appears to be regulated by at least two events: the first is independent of nitrogenase activity; the second is associated with nodule effectiveness.

Journal Article↗
PubMed1989

Biodegradation kinetics of linear alkylbenzene sulfonate in sludge-amended agricultural soils.

The kinetics of ultimate biodegradation (mineralization to CO2) of linear alkylbenzene sulfonate (LAS) were studied in sludge-amended agricultural soils for a series of pure chain length LAS homologs containing 10 to 14 carbon atoms in the alkyl chain. Degradation rates were measured by following the production of 14CO2 from uniformly 14C-ring-labeled material. In general, degradation of LAS was rapid in soil over a broad concentration range (0.1 to 10 times the expected environmental concentration) and demonstrated little variation among different homologs. Half-lives for mineralization of the benzene ring ranged from 18 to 26 days and were not significantly different for any homolog over the range of alkyl chain lengths tested. Half-lives measured for LAS degradation in these studies were comparable to values reported in the literature and also to values obtained for naturally occurring materials (stearic acid, cellulose) typically present in soil environments. On the basis of the results of the present studies and those of other investigators, it is concluded that soil environments exposed to LAS in sewage sludges contain microbial communities which can actively metabolize this material. Rates of biodegradation of the benzene ring, the final step in the LAS biodegradation pathway prior to complete mineralization, are also sufficient to prevent LAS from accumulating in soil environments.

Alkanesulfonates↗
PubMed1986

Kinetics of biodegradation of nitrilotriacetic acid (NTA) in an estuarine environment.

The effects of salinity and dissolved organic carbon (DOC) on the kinetics of biodegradation of nitrilotriacetic acid (NTA) were studied in a Canadian estuary with a prior history of NTA exposure. Kinetic parameters for degradation of 14C-labeled NTA, maximum velocity (Vmax) and first-order rate constant (k1), were estimated by nonlinear regression models from velocity and time-course plots, respectively. The distribution of bacteria with NTA-degrading capability was also determined at various salinities and DOC levels by the 14C-most-probable-number (14C-MPN) technique. In general, NTA degradation was rapid in estuarine water over the range of salinities and DOC levels tested. Mean Vmax and k1 values (+/- standard deviation) across several sampling periods averaged 4753 +/- 2849 ng liter-1 hr-1 and 0.32 +/- 0.19 day-1, respectively. The estimated half-life for NTA degradation in estuarine water, based on the mean k1 value, was approximately 2 days. Degradation rates for NTA were relatively insensitive to changes in salinity or DOC values, and neither of these two parameters had significant effects on NTA degradation at the microbial community or individual cell levels. Based on 14C-MPN results, the distribution of estuarine bacteria capable of degrading NTA was broad and not related to salinity or DOC levels. The NTA degraders appeared to be indigenous members of the estuarine microbial community and not wastewater-associated microorganisms.

Acetates↗
PubMed1986

Adaptation of aquatic microbial communities to quaternary ammonium compounds.

The effects of long-chain (C(12) to C(18)) quaternary ammonium compounds (QACs) on the density, heterotrophic activity, and biodegradation capabilities of heterotrophic bacteria were examined in situ in a lake ecosystem. Monoalkyl and dialkyl substituted QACs were tested over a range of concentrations (0.001 to 10 mg/liter) in both acute (3 h) and chronic (21 day) exposures. In general, none of the QACs tested had significant adverse effects on bacterial densities in either acute or chronic studies. However, significant decreases in bacterial heterotrophic activity were noted in acute studies at QAC concentrations from 0.1 to 10 mg/liter. Chronic exposure of lake microbial communities to a specific monoalkyl QAC resulted in an adaptive response and recovery of heterotrophic activity. No-observable-effect level in the adapted populations was >10 mg/liter. Chronic exposure also resulted in significant increases in the number and activity of bacteria capable of biodegrading the material. The increase in biodegradation capability was observed at low (microgram per liter) concentrations which are approximately the same as realistic environmental levels. In general, our studies indicated that exposure of lake microbial communities to QACs results in the development of adapted communities which are less sensitive to potential toxic effects and more active in the biodegradation of these materials.

Journal Article↗