This research evaluated dissolved air flotation (DAF) as a separation method for algae and phosphorus from municipal wastewater at the City of Logan, Utah Wastewater Reclamation Facility. DAF uses the supersaturation of air to raise suspended algae and other particles to the surface, where they can be easily removed. DAF, in conjunction with chemical coagulants and flocculants, can approach 95% algae and phosphorus removal. The algae removed using the DAF process will be used in the production of biofuels and bioplastics. A pilot DAF unit was used to determine the optimum alum dose for total phosphorus (TP) and algae removal. In addition, a bench-scale jar test unit was used to study the effects of various alum and polymer doses on removal efficiencies at different times of the day. An optimal alum dose was found to be 30 mg/L based on results from both the pilot and bench-scale units. No advantage to adding polymer was found. Algae removal efficiencies on the pilot DAF ranged from 68-70%, and the effluent algae concentration was reduced to 10 mg/L. Approximately 65% of the total phosphorus was removed, from 1.1 to 0.4 mg/L, which is low enough to meet regulations anticipated to be promulgated by the state of Utah. Using the assumption that the molar weight of algae is 3,550 g/mole, the molar ratio of Al/TSS was found to be 30.1 and the molar ratio of Al/TP was found to be 7.5. Extracellular polymeric substances (EPS) excreted by algal cells act as a natural flocculant and may allow for chemical usage to be minimized. Autoflocculation and bioflocculation, natural processes caused by EPS production and an increased pH level, were not observed to be a significant factor. The chemical dosing rates provide the City of Logan with basic operational parameters for a full-scale (15 million gallons per day) DAF plant, providing an effluent phosphorus level below 0.5 mg/L. The alum will cost $1,118 per day, with a daily electrical cost of approximately $149. This full-scale DAF plant would harvest 1,563 kg of algal biomass per day, with a cost per kilogram of algae at $0.81.

Investigates several hypotheses regarding the use of bioflocculation as a harvesting method for wastewater pond microalgae. Research performed on pilot-scale high rate ponds (HRPs) on the California Central Coast.

In recent years there has been growing interest in bio-mineral processing due to its low operating costs and its application in processing lean-grade ores. Bioprocessing is a good alternative to conventional hydrometallurgy process in mineral processing. In recent times microorganisms have been used as surface modifiers in processes such as froth flotation and flocculation. The surface properties of microbes and minerals such as zeta potential and surface hydrophobicity play a major role in determining adhesion of microorganisms to minerals and hence, the efficiency of flocculation and flotation. These properties also depend on solution conditions such as pH and ionic strength. Adhesion of microorganisms to mineral surfaces can alter the surface properties of the minerals. Such surface modification imparting hydrophobicity or hydrophilicity is used in flocculation and flotation of fine particles. In this research work the effect of ionic strength and pH in deteraiining the surface properties and hence adhesion of the bacterium Bacillus polymyxa to minerals such as hematite, quartz and coal has been studied in detail. The effect of the ionic strength and pH on the electrokinetics of the minerals and bacteria and its subsequent effect on adhesion and flocculation were investigated in detail. Contact angle measurements along with the zeta potential results were used to calculate the interaction energies between the mineral and the microorganism to establish a mechanism for the interaction. The following major conclusions can be drawn from this study. Results indicate that increase in the ionic strength significantly changes the zeta potential of hematite and bacteria without varying the isoelectric point. Increase in the ionic strength caused very little change in the zeta potential of quartz and coal. The adhesion of bacterial cells on to the minerals was found to be dependent on pH, ionic strength and conditioning time. Adhesion of bacterial cells was found to be more on hematite and coal when compared to quartz. The adsorption isotherms of Bacillus polymyxa cells with respect to all the three minerals were found to obey Langmuir isotherm. Flocculation studies demonstrated that the settling rate of hematite and coal was enhanced in presence of bacterial cells and electrolyte. However quartz settled much slower under the same conditions indicating that the quartz particles are being dispersed. Thus, selective flocculation of hematite and coal is possible which can be used in separating them from quartz effectively. The different components of total interaction energy arising from Lifshitz-van der Waal forces, acid/base forces and electrostatic forces were calculated using the van Oss approach. Calculation of the components of the acid base free energy showed that coal and hematite were hydrophobic compared to quartz and the bacterium. From total interaction energy calculation based on the extended DLVO theory, hematite and coal were found to have a net negative interaction energy in acidic pH values and hence attractive forces are predominant. Quartz was found to have a net repulsive energy at all the pH values at low ionic strengths but increase in ionic strength the interaction energy become attractive. The AGLW values of quartz was found to be attractive which is probably responsible for bacterial adhesion onto quartz.

Enzymes are extremely attractive biological catalysts in industrial conversion processes due to their ability to promote reactions with high enantioselectivity and in milder conditions than chemicals, as well as biological flocculants are advantageous for easy biodegradation and safety to living beings. Both are easily obtained from microorganisms that naturally inhabit environments where these molecules are necessary to favor their metabolism. In the present work, four bacterial isolates were used, two from the Atlantic Forest but cultivated for 40 years with sugarcane (LBPMA strains: APF.SG3-Isox and ACO.PR1-Isox), previously selected by high tolerance to agrochemicals, and two from agro-industrial sludge (LBPMA: BDLJ2 and BDL07) (Coruripe-AL, Brazil), sorted for producing polyhydroxyalkanoates and biosurfactants. Each lineage was submitted to biochemical and molecular tests for identification purposes and then submitted to qualitative / semi-quantitative tests (cultures in solid medium containing specific substrate) and quantitative (submerged cultures containing specific substrates) of cellulolytic, proteolytic, lipolytic activity and bioflocculant producer. Based on the morphological properties and analyzes of the 16S rDNA sequences, the isolates LBPMA-APF.SG3-Isox, LBPMA-ACO.PR1-Isox, LBPMA-BDLJ2 and LBPMA-BDL07 were identified respectively as B. megaterium, B. toyonensis, B. thuringiensis and B. pumilus, and all were able to present the studied activities. The maximum indices for protease and lipase activity in solid culture were obtained by B. toyonensis (IE = 4.55 and 2.41, respectively), while the maximum cellulolytic activity in solid medium (IE = 1.40) was obtained by B. pumilus. In liquid medium, the strains of B. pumilus and B. thuringiensis showed the highest cellulolytic activity in the first 48 h. The maximum lipolytic activity of the three isolates analyzed in submerged culture containing olive oil as a specific substrate (0.274 U. mL-1 for Bacillus thuringiensis LBPMA-BDLJ2, 0.450 U. mL-1 for Bacillus pumilus LBPMA-BDL07 and 0.552 U. mL-1 for Bacillus megaterium LBPMA- APF.SG3), also corresponded to the maximum cell growth at 72 h of incubation. Concerning proteolytic activity in submerged culture, B. thuringiensis showed the same maximum activity as B. pumilus, but in a shorter incubation period. All isolates secreted the tested enzymes and bioflocculat at the constant temperature of 37 ± 1 ° C, without changes in the pH over time. The flocculant activity increased with the incubation time, initially reaching the maximum value of 57% at 72 h for B. pumilus culture, while for B. thuringiensis, B. toyonensis and B. megaterium this was 33%, 21% and 34%, respectively, after 24 h of incubation. These initial results stimulate the need for optimization of the pH (3, 5, 7 and 9) and nitrogen [Urea, Peptone and (NH4)2SO4] and carbon sources (Maltose, Sucrose and Glucose) variables of the medium for flocculation using such strains of the Bacillus genus. Sucrose and Maltose were the best sources of carbon, whilst Urea was the prefered source of nitrogen for two of the tested isolates (B. pumilus and B. toyonensis), followed by (NH4)2SO4 (B . megaterium) and Peptone (B. thurigiensis). The best pH values for the production of bioflocculant were 5.0 (B. toyonensis e B. thuringiensis) and 3.0 (B. pumilus e B. megaterium). The FTIR-ATR spectra of the extracted flocculants revealed the presence of the carboxyl, hydroxyl and methoxy functional groups as responsible for the flocculant activity of the studied strains, thus being characterized as polysaccharides. / Conselho Nacional de Desenvolvimento Científico e Tecnológico / Enzimas são catalisadores biológicos extremamente atrativos em processos de conversão industrial devido a suas habilidades em promover as reações com alta enantio-seletividade e em condições mais brandas que os químicos, da mesma forma que floculantes biológicos são vantajosos pela fácil biodegradação e segurança aos seres vivos. Ambos são facilmente obtidos a partir de microrganismos que naturalmente habitem ambientes onde tais moléculas se fazem necessárias para favorecer seu metabolismo. No presente trabalho, foram utilizados quatro isolados bacterianos, dois provenientes de solo de Mata Atlântica (linhagens LBPMA: APF.SG3-Isox e ACO.PR1-Isox), previamente selecionados pela alta tolerância a agroquímicos, e dois de lodo agroindustrial (linhagens LBPMA: BDLJ2 e BDL07) (Coruripe-AL, Brasil), triados por produzirem polihidroxialcanoatos e biosurfactantes. Cada linhagem foi submetida a testes bioquímicos e moleculares com fins de identificação e então, submetidas a testes qualitativos/semi-quantitativos (culturas em meio sólido contendo substrato específico) e quantitativos (culturas submersas contendo substratos específicos) de atividade celulolítica, proteolítica, lipolítica e produtora de biofloculante. Baseado nas propriedades morfológicas e análises das sequências 16S rDNA, os isolados LBPMA-APF.SG3-Isox, LBPMA-ACO.PR1-Isox, LBPMA-BDLJ2 e LBPMA-BDL07 foram identificados respectivamente como B. megaterium, B. toyonensis, B. thuringiensis e B. pumilus, e todos foram capazes de apresentar as atividades estudadas. Os índices máximos para atividade proteásica e lipásica em cultura sólida foram obtidos por B. toyonensis (IE= 4,55 e 2,41, respectivamente). Para atividade celulásica máxima, B. pumilus foi o isolado mais eficiente em meio sólido (IE=1,40), e em meio líquido, as estirpes de B. pumilus e B. thuringiensis apresentaram a máxima atividade nas primeiras 48 h. A atividade lipolítica máxima dos três isolados analisados em cultura submersa contendo azeite de oliva como substrato específico (0,274 U. mL-1 para Bacillus thuringiensis LBPMA-BDLJ2, 0,450 U. mL-1 para Bacillus pumilus LBPMA-BDL07 e 0,552 U. mL-1 para Bacillus megaterium LBPMA- APF.SG3), correspondeu também ao máximo crescimento celular às 72 h de incubação. Já em relação à atividade proteolítica em cultura submersa, B. thuringiensis apresentou a mesma atividade máxima que B. pumilus, porém num período menor de incubação. Todos os isolados secretaram as enzimas investigadas como também biofloculantes, ambos na temperatura constante de 37 ± 1 °C, sem alterações de pH ao longo do tempo. A atividade floculante aumentou com o tempo de cultivo, a princípio, chegando ao valor máximo de 57 % às 72 h para as amostras de B. pumilus, enquanto para B. thuringiensis, B. toyonensis e B. megaterium foram de 33 %, 21 % e 34 % (respectivamente), após 24 h de incubação. Estes resultados obtidos estimularam a otimização das variáveis pH (3, 5, 7 e 9), fontes de nitrogênio [Uréia, Peptona e (NH4)2SO4] e carbono (Maltose, Sacarose e Glicose) do meio, para melhoria das taxas de floculação por tais linhagens do gênero Bacillus. A Sacarose e a Maltose apresentaram-se como as melhores fontes de carbono, enquanto Uréia foi a fonte preferencial de nitrogênio para dois dos isolados testados (B. pumilus e B. toyonensis), seguido de (NH4)2SO4 (B. megaterium) e Peptona (B. thuringiensis). Os melhores valores de pH para a produção de biofloculante foram 5,0 (B. toyonensis e B. thuringiensis) e 3,0 (B. pumilus e B. megaterium). Os espectros de FTIR-ATR dos floculantes extraídos revelaram a presença dos grupos funcionais carboxilo, hidroxilo e metoxilo como responsáveis pela atividade floculante das estirpes estudadas, sendo assim caracterizados como polissacarídeos.

ABSTRACT IMPROVED MICROALGAL BIOMASS HARVESTING USING OPTIMIZED ENVIRONMENTAL CONDITIONS AND BACTERIAL BIOFLOCCULANTS DEREK CONTE MANHEIM The cost and energy balance of microalgae biofuel production is sensitive to the algae harvesting method, among many other factors. Bioflocculation and settling of suspended microalgae cultures is a harvesting method with potentially low cost and energy input. However, bioflocculation (the spontaneous flocculation of algal cells without chemical addition) has not been a reliable process with cultures grown in ponds. To provide insights to help improve algae settling, factors affecting the settling of algae were investigated in the laboratory using pure cultures of two common microalgae species: Scenedesmus sp. and Chlorella vulgaris. Bioflocculation of these algae was studied with and without the addition of bioflocculants produced by the bacterium, Burkholderia cepacia, to improve settling efficiencies. The bioflocculant produced by this bacterium was used in two different forms: a cell suspension including capsular and dissolved extracellular polymeric substance (EPS) components of B. cepacia, and dialyzed filtrate of the bacterial culture (only dissolved EPS fraction). The effects of algal growth phase, mixing time, bioflocculant dose, and environmental conditions such as pH and nutrient deprivation of bacterial bioflocculant cultures on settling of the algae species were studied. Settling characteristics were different for the two algae cultures, and their settling was affected differently by the many factors studied. Scenedesmus settling was best in later growth stages, while Chlorella settled much better in early growth phases. Addition of B. cepacia cells as a bioflocculant improved settling of Scenedesmus, with the greatest effect during mid to late exponential growth of the Scenedesmus. In contrast, addition of B. cepacia filtrate as a bioflocculant best improved Chlorella settling during stationary growth of Chlorella. Longer mixing times (contact time between the algae cells and bacterial bioflocculant) improved the settling of Scenedesmus, while Chlorella settled better with a shorter mixing time. Reducing the pH to 3 (a typical isoelectric point for microalgae) improved the settling of both algae cultures, with and without bioflocculant addition. Increasing the pH to 11 autoflocculated Scenedesmus cultures, but not Chlorella cultures, at early growth stages. EPS produced by the algae, bacteria, and wastewater organisms was quantified using dialysis separation followed by total organic carbon (TOC) analysis. Wastewater organisms were included because wastewater is a potential growth medium for biofuel algae. Improved settling of both species of algae depended on both the quantity and type of EPS (dissolved or capsular) produced by both the bacterial bioflocculant, and the algae themselves. Scenedesmus settled the best during late growth phases while its own EPS production was high, and combined EPS (capsular and dissolved) from B. cepacia improved settling at a higher dosage of bacterial cells to algae (1:2 B. cepacia cells to algae cells). Since Chlorella settling was not improved at later growth stages when its own EPS production was greatest, it appears that Chlorella’s settling rate was less affected by the production of its own EPS. For Chlorella, B. cepacia EPS addition (capsular and dissolved) was effective only in low doses (1:6 B. cepacia cells to algae cells). Settling results with the addition of bacterial bioflocculants with the pure algae cultures were compared to settling results of lab experiments with algae pondwater sampled from high-rate algae ponds (HRAPs). These algae samples were used to test the addition of return activated sludge (RAS) to improve settling. RAS addition improved the settling of Chlorella, which was the dominant algae species in the HRAP during the time of this study, at two different doses (a ratio of RAS to algae pond water of 1:3 and 1:6). Nutrient deprivation of B. cepacia cells before use as a bioflucculant was found to improve settling for Scenedesmus, especially during early phases of growth when EPS production of Scenedesmus was low. The EPS produced by the starved bacterial cells was about 30% greater than that produced by cultures which were not nutrient-limited. For the bacterial cultures, EPS production peaked at mid stationary phase for non-starved cultures and during early stationary phase for starved cultures. Chlorella settling improved in early growth with starved bacterial cell addition and in later growth with non-starved bacterial cell addition. These results suggest that the settling of microalgae can vary dramatically by species and that the settling of different species is affected differently by growth phase and environmental conditions. In addition, species of algae respond differently to addition of bacterial bioflocculants. Given the dramatically different settling behavior of the two species of algae used in this research, more research should be directed to studying settling of other microalgal species. Based on this research, the use of bacterial bioflocculants is promising for improving algae settling and may contribute to the development of a reliable, low cost harvesting process for commercial biofuel production from microalgae.

Bioflocculation of Wastewater Treatment Pond Suspended Solids Louis Lefebvre Wastewater treatment lagoons and high rate algae ponds (HRAPs) can provide cost effective wastewater treatment, but they commonly have high effluent concentrations of total suspended solids (TSS). In this thesis algae pond effluent was treated in a beaker testing apparatus (mixed and aerated) with various mixtures of activated sludge and primary effluent simulating differing activated sludge aeration basin compositions then was allowed to settle to assess settleability. Conventionally, microalgal suspended solids are removed by chemical coagulation followed by separation methods that often have a high cost relative to the low cost lagoon or HRAP system where the solids were produced. This separation step is often cost prohibitive or operationally complex for municipalities or too energy intensive for application in algae biofuels production. This research investigates using a small amount of activated sludge material to promote bioflocculation of algae in pond effluent. It was hoped that the findings may demonstrate a path for municipalities to keep their lagoons, while increasing capacity and improving treatment without excessive cost or complexity. Experiments were conducted on microalgae samples from a pilot-scale HRAP and activated sludge and primary effluent samples from a local municipal wastewater plant. The samples were placed in a mixing apparatus and allowed to settle for a given period of time, after which TSS was analyzed for settleability. The experiments investigated the effect of various lab-scale activated sludge reactor operational schemes by varying the volumes (and masses) of activated sludge, algae-rich water, and activated sludge in the beaker. Results in the sorption test (tests with only activated sludge and algae-rich water) demonstrated algae pond effluent treated with activated sludge concentrations of 3000 mg/L or greater produced final effluent TSS concentrations near discharge requirements (40-50 mg/L) with only 30 minutes of settling and without addition of primary effluent. However, such high activated sludge concentrations are not feasible at full scale. Furthermore, beakers with activated sludge concentrations greater than 3000 mg/L reduced TSS concentrations by more than 150 mg/L with only 30 minutes of settling and without addition of primary effluent. Results in the aerobic beaker tests (tests with primary effluent, activated sludge, and algae-rich water) showed greater than 200 mg/L TSS removal and final effluent TSS concentration less than 30 mg/L was achieved using activated sludge to primary effluent volumetric ratios of 1:1 and greater which corresponded to activated sludge concentrations of 730 mg/L and greater. Activated sludge concentrations of 730 mg/L may not be feasible at full scale. This report shows that a PETRO-like process is effective in lowering wastewater pond suspended solids, however not to typical discharge standards.

Two sets of triplicate pilot algal raceway ponds (1000-L, 0.30-m deep, paddle wheel mixed) were operated for 14 months at a California wastewater treatment plant to treat wastewater and generate algal-bacterial biomass as biofuel feedstock. Two experiments were run to determine the effect on biomass productivity of (1) hydraulic residence time (HRT: 2, 3, 4, or 4.5 days) and (2) feeding schedule (18 small pulses during 8 AM-4 PM [diurnal] versus 20 large pulses during 4 AM-12 AM [diel]). The target productivity was at least 20 g volatile suspended solids per m2 of pond per day. Additional output variables were followed during the experiments: treatment performance and the effectiveness of biomass harvesting though bioflocculation. Productivity was consistently higher in ponds with a 2-d HRT versus longer HRTs. Average productivity for the 2-d HRT ponds and the variable-HRT ponds (3.6-d average HRT) were 30.1 and 23.4 g/m2-d, respectively. Productivity data collected during the feed regime experiment were highly variable, and average productivities were the same at 26 g/m2-d. During both experiments, both pond sets exceeded the target of 20 g/m2-d on an annual basis. During the hydraulic residence time experiment, the average pond productivity throughout the HRT experiment for the 2-d HRT and 3-d HRT ponds were 30.1 and 23.4 g/m2-d, respectively. Settling efficiency was high for both 2- and 3-d HRT ponds with average turbidity removal of 87-89%. However, total ammonia nitrogen (TAN) concentrations in the 2-day HRT pond effluent were 50-94% higher than in the 3-d HRT pond effluents, although effluent TAN concentrations in both ponds were approximately the same during mid-summer. Furthermore, effluent biochemical oxygen demand (BOD5) concentrations were similar, with the supernatant of Imhoff cones settled for 24 hours containing 24-27 mg/L BOD5 (81-92% removal). In general, the 3-d HRT ponds provided better treatment than the 2-d HRT ponds. During the feeding regime experiment, no productivity or BOD5 removal differences were evident. However, the 3-d HRT ponds had consistently 8 mg/L more effluent TAN than the 2-d HRT ponds.

Extracellular polymeric substances (EPS) in activated sludge are known to account for the flocculent nature of activated sludge. Extensive studies over the last few decades have attempted to extract and characterize activated sludge EPS, but a lack of agreement between studies has also been quite common. The molecular makeup of EPS has, however, remained nearly unexplored, leaving their identity, function, and fate over various stages in the activated sludge system mainly unknown. In spite of their critical involvement in bioflocculation and long history of related research, our understanding of EPS is still greatly limited and better elucidation of their composition and structure is needed. The hypothesis of this research was that activated sludge floc contains different fractions of EPS that are distinguishable by their association with certain cations and that each fraction behaves differently when subjected to shear, aerobic digestion, anaerobic digestion and other processes. In order to examine this floc hypothesis, the research mainly consisted of three sections: 1) development of EPS extraction methods that target cations of interest (divalent cations, especially calcium and magnesium, iron, and aluminum) from activated sludge; 2) molecular investigations on activated sludge EPS using metaproteomic analyses, comprising sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and protein identification by liquid chromatography tandem mass spectrometry (LC/MS/MS), and hemaagglutination (HA)/HA inhibition assays; and 3) investigating the fate of EPS in sludge digestion using SDS-PAGE. Evaluation of prior research and data from preliminary studies led to the development of the three extraction methods that were used to target specific cations from activated sludge and to release their associated EPS into solution. These methods are the cation exchange resin (CER) procedure for extracting Ca²⁺+Mg²⁺, sulfide extraction for removing Fe, and base treatment (pH 10.5) for dissolving Al. The cation selectivity in the three extraction methods, the composition of EPS (protein/polysaccharide), amino acid composition, and a series of sequential extraction data established initial research evidence that activated sludge EPS that are associated with different cations are not the same. SDS-PAGE was successfully applied to study extracellular proteins from several sources of both full- and bench-scale activated sludges. The three extraction methods led to different SDS-PAGE profiles, providing direct evidence that proteins released by the three methods were indeed different sludge proteins. Another important outcome from this stage of research was finding the similarity and differences of extracellular proteins between different sources of activated sludge. SDS-PAGE data showed that many of CER-extracted proteins were well conserved in all the sludges investigated, indicating that a significant fraction of Ca²⁺ and Mg²⁺-bound proteins are universal in activated sludge. On the other hand, protein profiles resulting from sulfide and base extraction were more diverse for different sludges, indicating that Al and Fe and their associated proteins are quite dynamic in activated sludge systems. Protein bands at high densities were analyzed for identifications by LC/MS/MS and several bacterial proteins and polypeptides originating from influent sewage were identified in this study. This was also thought to be the first account of protein identification work for full-scale activated sludge. The analysis of SDS-PAGE post sludge digestion revealed that CER-extracted proteins remained intact in anaerobic digestion while they were degraded in aerobic digestion. While the fate of sulfide-and base-extracted proteins in aerobic digestion was not as clearly resolved, their changes in anaerobic digestion were well determined in this research. Sulfide-extracted protein bands were reduced by anaerobic digestion, indicating that Fe-bound EPS were degraded under anaerobic conditions. While parts of base-extracted proteins disappeared after anaerobic digestion, others became more extractable along with the extraction of new proteins, indicating that the fate of base-extractable proteins, including Al-bound proteins, is more complex in anaerobic digestion than CER-extracted and sulfide-extracted proteins. These results show that Ca²⁺+Mg²⁺, Fe³⁺, and Al³⁺ play unique roles in floc formation and that each cation-associated EPS fraction imparts unique digestion characteristics to activated sludge. Finally, since a considerably different cation content is quite common for different wastewaters, it is postulated that this variability is one important factor that leads to different characteristics of activated sludge and sludge digestibility across facilities. The incorporation of the impact of cations and EPS on floc properties into an activated sludge model might be challenging but will assure a better engineering application of the activated sludge process. / Ph. D.

Studies were conducted to determine the role of bioflocculation in the activated sludge unit processes. Laboratory and full-scale studies revealed that bioflocculation is important in determining settling, dewatering, effluent and digested sludge properties (activated sludge properties) and may be vital to the function of all processes related to the above properties. In these studies, it was shown that divalent cations such as calcium and magnesium improved activated sludge properties, whereas monovalent cations such as sodium, potassium and ammonium ions were detrimental to these properties. The divalent cations promoted bioflocculation through charge bridging mechanisms with negatively charged biopolymers (mainly protein and polysaccharide). It was found that oxidized iron plays a major role in bioflocculation and determination of activated sludge properties through surface interactions between iron and biopolymers. Oxidized iron was effective in removing colloidal biopolymers from solution in coagulation and conditioning studies. The research included experiments evaluating effects of potassium and ammonium ions on settling and dewatering properties; effects of magnesium on settling properties; effects of sodium, potassium, calcium and magnesium on effluent quality; effect of solids retention time on effluent quality; and evaluation of floc properties during aerobic and thermophilic digestion. A floc model is proposed in which calcium, magnesium and iron are important to bioflocculation and the functionality of aeration tanks, settling tanks, dewatering equipment and aerobic or anaerobic digesters. It is shown that activated sludge floc properties affect wastewater treatment efficiency. / Ph. D.

The possible effects of changes in the feeding pattern on activated sludge properties related to bioflocculation have been analyzed in lab scale sequencing batch reactors (SBR) in order to determine if these changes in effluent water quality and settling and dewatering properties are significant, so they can be considered in future studies or if they can be recommended as crucial when operating and designing wastewater treatment plants. The activated sludge process is widely used to treat wastewater from both industrial and municipal sources. Biomass from industrial facilities containing high monovalent to divalent ion content usually settles poorly, which leads to low quality effluents that fail to meet environmental requirements. Therefore, the combined effect of feeding pattern plus the addition of sodium to activated sludge reactors was studied in this experiment. A series of SBRs were operated at different sodium concentrations that ranged from 1.5 - 15 meq/L and different feeding times that ranged from 1 minute to 4 hours. Biomass samples were taken from each reactor to study the settling and dewatering properties and effluent samples were used to analyze the amount of organic matter and exocellular polymeric substances present due to deflocculation. As expected, the changes in feeding strategies affected all of the properties measured. When the feeding time was maintained low (pulse feed) the effluent quality and settling properties were the best. As the feeding time was increased the effluent quality, settling, and dewatering properties increased suggesting that the way in which the reactors were fed affected the overall bioflocculation process. The causes of the high deflocculation observed are not well understood, but data suggest that a microbial community change could have affected exocellular biopolymers which are believed to play an important role on bioflocculation. This research demonstrates the importance of the interaction between cation content and feeding pattern when operating a wastewater treatment plants and when reporting lab-scaled results related to settling and bioflocculation. / Master of Science