Characterization of flocculation for cell removal from fermentation broth via polyelectrolyte addition is commonly based on qualitative methods such as physical appearance of the floc. The use of zeta potential as a quantitative measure of floc character was evaluated as an indicator of optimal polymer addition. Zeta potential was found to increase with increasing cationic polyelectrolyte dosage, but never reached zero regardless of the total amount of polymer added, indicating flocculation occurs at least partially through a bridging type mechanism. Experiments were conducted using various polymer concentrations (25-75 g/L) and dosing methods (batch, incremental and continuous addition) that resulted in variable overall polymer requirements to achieve optimum flocculation. Zeta potential was found to be constant at optimal floc character regardless of the total amount of polymer added, polymer concentration, or method of polymer addition. Experiments with two additional types of fermentation broth also showed characteristic zeta potentials at optimal flocculation. Polymer requirements to achieve a particular floc character can vary greatly, depending on polymer dosing conditions and fermentation batch. The effect of polymer dosing conditions on the polymer requirement to obtain optimal floc character was evaluated. Polymer dosing method and calcium concentration were both found to have a significant effect (P < 0.0001) with continuous polymer addition and high calcium concentration requiring less polymer than did batch polymer addition and low calcium concentration, respectively. Polymer dosing concentration did not significantly affect polymer requirement for optimal flocculation.

Water distribution systems are complex environments frequently containing corroded iron pipes and biofilms. To thoroughly understand the fate of halogenated disinfection byproducts (DBPs) in these systems, two degradation processes were investigated: abiotic degradation (i.e. hydrolysis and reductive dehalogenation) and biodegradation. DBPs were selected from 6 different compound classes representing both regulated DBPs (i.e. trihalomethanes or THMs, and haloacetic acids or HAAs) and non-regulated or "emerging" DBPs. Batch experiments were conducted to investigate the pathways and kinetics of DBP degradation. As expected, the relative importance of hydrolysis, abiotic reductive dehalogenation, and biodegradation depends on the DBP structure and on the environmental conditions (i.e. pH, temperature, dissolved oxygen, Fe minerals present, bacteria present, etc.). From our results, chloropicrin (i.e. trichloronitromethane) and most brominated DBPs are highly susceptible to abiotic reductive dehalogenation, trichloracetonitrile and trichloropropanone are the most susceptible to hydrolysis, and HAAs are readily biodegraded under aerobic conditions. Knowledge of DBP degradation mechanisms and rates in distribution systems is important for selecting DBP monitoring locations, modeling DBP fate, and for predicting exposure to these compounds. Such information could also be useful for developing treatment systems for DBP removal. 334

Halonitromethanes (HNMs) are a class of halogenated disinfection byproducts formed upon the addition of chlorine to water containing organic matter. Batch experiments were performed to investigate the reaction pathways and kinetics of three HNMs (chloropicrin or trichloronitromethane [TCNM], dichloronitromethane [DCNM], and chloronitromethane [CNM]) with zero-valent iron (Fe0). All three compounds reacted rapidly in the presence of Fe0 (1.8-4.4 g/L) with methylamine (MA) as the final product. The geometric surface area-normalized rate constants decreased with decreasing halogenation: TCNM (301 L/[h-m2]) > DCNM (153 L/(h-m2)) > CNM (45.9 L/[h-m2]). Nitromethane, an intermediate species, rapidly reacted to form MA (302 L/[h-m2]). These reactions all experienced some degree of mass transfer limitation (9-73%). The average carbon and chlorine mass balances for TCNM were >85%, indicating that the major reaction products were recovered. The degradation of TCNM and DCNM proceeded via the parallel reaction pathways of hydrogenolysis and alpha-elimination. For TCNM, 60.7 +/- 8.7% of reaction proceeded via hydrogenolysis and 39.3 +/- 6.4% via alpha-elimination. Knowledge of HNM reaction pathways and kinetics in the presence of Fe0 may be useful for predicting the fate of these compounds in drinking water distribution systems containing cast or ductile iron pipe and for developing treatment systems for HNM removal from water.

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Association of extracellular protein product with flocculated cells reduces product yield. Here, partitioning of the enzyme subtilisin between the liquid and polyelectrolyte-flocculated and sedimented Bacillus increased as the polymer dosage was increased beyond that necessary to obtain optimum floc character (brain floc) for cell removal by centrifugation. Partitioning to the cell floc is partly physical entrapment at all polymer dosages; however, at higher levels there is also direct interaction between the polyelectrolyte and enzyme. Enzyme loss was not likely due to pH denaturation during the flocculation process because conditions were within the stable pH range of the enzyme. The direct interaction between polyelectrolyte and enzyme was characterized through turbidimetric titrations and partitioning studies. Neither changes in the polymer feed concentration nor the method of polymer addition reduced the enzyme loss at dosages optimal for cell removal.