Browsing by Subject "EPS"
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Item Characterization of marine exopolymeric substance (EPS) responsible for binding of thorium (IV) isotopes(Texas A&M University, 2005-08-29) Alvarado Quiroz, Nicolas GabrielThe functional group composition of acid polysaccharides was determined after isolation using cross-flow ultrafiltration, radiolabeling with 234Th(IV) and other isotopes, and separation using isoelectric focusing (IEF) and polyacrylamide gel electrophoresis (PAGE). Phosphate and sulphate concentrations were determined from cultured bacterial and phytoplankton colloid, particulate and colloidal samples collected from the Gulf of M??xico (GOM). Characterization of the 234Th(IV)-binding biomolecule was performed using ion chromatography (IC), and gas chromatography-mass spectrometry (GC-MS). Radiotracer experiments and culture experiments were conducted in determining the binding environment of the 234Th(IV)-binding ligand (i.e., sorption onto suspended particles), as well as the origin of the ligand in seawater systems. In all samples, 234Th(IV) isoelectric focusing profiles indicated that 49% to 65% of the 234Th(IV) labeled EPS from Roseobacter gallaeciensis, Sagittula stellata, Emiliania huxleyi, Synechococcus elongatus and GOM Station 4-72m was found at a pHIEF of 2 in the IEF spectrum. The carboxylic acid group appeared at the same pHIEF as 234Th(IV) for EPS from Roseobacter gallaeciensis, Emiliania huxleyi, Synechococcus elongatus and GOM colloidal organic matter sample. The phosphate group appeared at the same pHIEF as 234Th(IV) for EPS from Roseobacter gallaeciensis, and Synechococcus elongatus sample. The sulphate group was found at the same pHIEF as 234Th(IV) for EPS from S. elongatus and GOM colloidal organic matter sample. The total polysaccharide content was only 14% and 8%, uronic acids were approximately 5.4% and 87.1%, and total protein content was 2.6% and 6.2% of total carbon content of Sagittula stellata and Synechococcus elongatus, respectively. Monosaccharides identified in both Sagittula stellata and Synechococcus elongatus were galactose, glucose, and xylose in common. In addition, Sagittula stellata contained mannose and Synechococcus elongatus had galactoglucuronic acid. Thus, depending on the species, the size, structural composition, and functional groups of the 234Th(IV)-binding, acidic polysaccharides will vary. From these observations, it is concluded that the steric environment and not necessarily the exact functional group might actually be responsible for thorium-234 complexation to macromolecular organic matter. This research helped to improve our understanding of the observed variability in POC/234Th ratios in the ocean and provided insights into factors that regulate organic carbon export fluxes.Item The effect of nutrient limitations on the production of extracellular polymeric substances by drinking-water bacteria(2013-05) Evans, Ashley Nichole; Kirisits, Mary JoBiological filtration (biofiltration) of drinking-water is gaining popularity due the potential for biodegradation of an array of contaminants not removed by traditional drinking-water processes. However, previous research has suggested that biomass growth on biofilter media may lead to increased headloss, and thus, greater energy and water requirements for backwashing. Research has suggested that the main cause of headloss might be due to extracellular polymeric substances (EPS) rather than the bacterial cells themselves. As EPS production has been shown to increase under nitrogen- and phosphorus-limited or -depleted conditions, the goal of this research was to add to the body of knowledge regarding biofiltration by studying the relationship between EPS production and nutrient limitations in drinking-water. Batch experiments with a synthetic groundwater were run with a mixed community of drinking-water bacteria under nutrient-balanced (a molar carbon to nitrogen to phosphorus ratio [C:N:P] of 100:10:1), nutrient-limited (e.g., C:N:P of 100:10:0.1), and nutrient-depleted conditions (C:N:P of 100:0:1 or 100:10:0). After 5 days, growth was measured as the optical density at 600 nanometers (OD600), and the concentrations of free and bound carbohydrates and proteins, the main components of EPS, were measured. In batch experiments with 2.0 and 0.2 g/L as carbon (mixture of acetic acid, mannitol and sucrose) increases in EPS production per OD600 and decreases in growth were noted under nutrient-depleted conditions. When the same experiments were conducted with a pure culture of Bacillus cereus, bound polysaccharides normalized to OD600 increased under nitrogen- and phosphorus-depleted conditions. Since previous research suggested that Bradyrhizobium would be an important player in EPS production in drinking-water biofilters, similar batch experiments were conducted with Bradyrhizobium. However, due to experimental challenges with Bradyrhizobium japonicum USDA 110, differences in EPS production under nutrient limitations could not be reliably assessed. Additional work is required with Bradyrhizobium. Recommendations for future work include the replication of these batch conditions in steady-state chemostats containing biofilm attachment media and in bench-scale columns. Additionally, future work should include experiments at carbon concentrations as low as 2 mg/L to match typical carbon concentrations in drinking-water biofilters.Item Factors Affecting Carbohydrate Production and Loss in Salt Marsh Sediments of Galveston Bay(2010-10-12) Wilson, Carolyn E.Benthic microalgae (BMA) living within the surface sediment of salt marshes are highly productive organisms that provide a significant proportion of organic carbon inputs into estuarine systems. BMA secrete extracellular carbohydrates in the form of low molecular weight carbohydrates and extracellular polymeric substances (EPS) as they migrate within the sediment. EPS plays an important role in the structure and function of BMA biofilms in shallow-water systems as EPS affects habitat structure, stabilizes the sediment, reduces sediment erosion, and is a carbon source for organisms. This study looked at the effect of nutrients and carbohydrate additions on BMA biomass, bacterial biomass, carbohydrate production, and glycosidase activity in the surface 5 mm of intertidal sediment in a subtropical salt marsh (Galveston Bay, Texas). Nitrogen and phosphorus were added to cores collected from the salt marsh and incubated in the lab over four days. Very little change was seen in the biomass of the benthic microalgae or in the different carbohydrate fractions with the added nutrients. The mean chlorophyll a concentration was 13 +/- 5 ug g-1 sediment, the mean saline extractable carbohydrate concentration was 237 +/- 113 ug g-1 sediment, and the mean EPS concentration was 48 +/- 25 ug g-1 sediment. The chlorophyll a and saline extractable carbohydrate concentrations initially decreased over the first 24 hours, but then increased over the rest of the experiment, indicating a possible species compositional shift in the BMA. With no major response with nutrient additions, it is likely that a different environmental factor is limiting for the growth of the benthic microalgae, and therefore the production of sEPS, in this salt marsh. A series of experiments was conducted in situ by adding glucose, alginic acid, and phosphorus to sediment within experimental plots. Samples were taken periodically over three to seven days to determine the biomass of the microbial community, enzyme activities and kinetics, and changes in the concentrations of several sediment carbohydrate pools. u-glucosidase activities (15 +/- 3 nmol g-1 h-1) were significantly higher than u-xylosidase (6 +/- 2 nmol g-1 h-1) and u-galactosidase (8 +/- 2 nmol g-1 h-1) activities within the sediment, and there was no suppression of u-glucosidase activity measured with the glucose addition. These data represent the first measurement of u- xylosidase and u-galactosidase activity in intertidal sediment dominated by BMA. Although preliminary experiments suggested a possible phosphorus limitation within the sediment, there was little change in the bacteria abundance or the benthic microalgae biomass when phosphorus was added in situ. This study begins to illustrate the dynamics of carbohydrate production and loss in this salt marsh, and the ability for the microbial community in the salt marshes of Galveston Bay to adjust to the nutrient and carbohydrate treatments.