Using a pulsed electric field as a pretreatment for improved biosolids digestion and methanogenesis.

Researchers tested using pulsed electric field (PEF) technology to enhance conversion of organic solids material in waste activated sludge (WAS) and pig manure to soluble and colloidal forms, which are more bioavailable for methane production during subsequent anaerobic digestion. Operating parameters were varied from 1.1 to 19.8 kWhr/m3 to show the influence amount of treatment has on soluble chemical oxygen demand (SCOD), small colloidal solids, and methane production via the biochemical methane potential test. When PEF treatment exceeded a threshold, which was approximately 10 kWhr/m3, focused pulsed treatment solubilized approximately 10% of the total COD, increasing SCOD from as low as 20 mg/L to more than 1000 mg/L. The process also disrupted a larger portion of the volatile suspended solids (VSS) to form small colloids not measured by the VSS assay (between 0.2 and 1.2 microm). The effects increased the biological methane potential of the samples significantly: by 80% for pig manure and 100% for WAS after 25 to 30 days. These results support the conclusion that PEF pretreatment before anaerobic digestion has the potential to significantly improve digester performance, resulting in added methane production and decreased residual biosolids.

Thermodynamic evaluation on H2 production in glucose fermentation.

The normal maximum H2 yield in mesophilic biohydrogen (bioH2) fermentation is approximately 2 mol of H2/(mol of glucose). Thermodynamics could be the most fundamental control for bioH2 formation, since proton reduction is strongly energy consuming (+79.4 kJ/(mol of H2)). However, most of the electron equivalents in glucose do not accumulate in H2 but in a range of organic acids and alcohols. Thus, evaluating the hypothesis of thermodynamic control requiresthe full stoichiometry of the fermentation. We carried out batch bioH2 reactions with a range of pH values that yielded H2 yields from 0 to approximately 2 mol of H2/(mol of glucose). We constructed complete electron equivalent(e(-) equiv) balances for high or low H2 yield by measuring all e(-) sinks. The highest H2 yield occurred with pH approximately 4 and was coincident with major butyrate accumulation; ethanol or lactate correlated to reduced H2 yields at pH 7 and 10, respectively. Although the Gibb's free energies for all overall reactions were similar (-10.6 to -11.2 kJ/(e(-) equiv)), thermodynamics controlled the H2-producing reaction coupled to ferredoxin; this reaction was favorable at acidic pH but thermodynamically blocked at pH 10. Also, butyrate formation was the most thermodynamically favorable reaction that produced ATP after glycolysis.

Adhesion characteristics of two Burkholderia cepacia strains examined using colloid probe microscopy and gradient force analysis.

Colloid probe atomic force microscopy (CP-AFM) was used to investigate two strains of Burkholderia cepacia in order to determine what molecular scale characteristics of strain Env435 make it less adhesive to surfaces than the parent strain, G4. CP-AFM approach curves analyzed using a gradient force method showed that in a high ionic strength solution (IS=100 mM, Debye length=1 nm), the colloid probe was attracted to the surface of strain G4 at a distance of approximately 30 nm, but it was repelled over a distance of 25 nm when approaching strain Env435. Adhesion forces measured under the same solution conditions during colloid retraction showed that 1.38 nN of force was required to remove the colloid placed in contact with the surface of strain G4, whereas only 0.58 nN was required using strain Env435. At IS=1mM (Debye length=10nm), the attractive force observed with G4 was no longer present, and the repulsive force seen with Env435 was extended to approximately 250 nm. The adhesion of the bacteria to the probe was much less at low IS solution (1 mM) than at high IS (100 mM). The greater adhesion characteristics of strain G4 compared to Env435 were confirmed in column tests. Strain G4 had a collision efficiency of alpha=0.68, while strain Env435 had a much lower collision efficiency of alpha=0.01 (IS=100 mM). These results suggest that the reduced adhesion of strain Env435 measured in column tests is due to the presence of high molecular weight extracellular polymeric substances that extend out from the cell surface, creating long-range steric repulsion between the cell and a surface. Adhesion is reduced as these polymers do not appear to be "sticky" when placed in contact with a surface in AFM tests.

Transport of rodlike colloids through packed beds.

The effect of colloid shape on filtration rates in porous media was examined by constructing particles with different aspect ratios and measuring their retention in packed beds. Spherical polystyrene latex microspheres (1.0-microm diameter) were heated, stretched to the desired aspect ratio (2:1 and 3:1, with a 1:1 control), and quickly cooled. These particles were injected into minicolumns containing glass beads (40-microm diameter) in solutions at two different ionic strengths (IS = 1 and 100 mM). The measured retentions increased with aspect ratio in both IS solutions. The zeta-potentialsfor all three aspect ratios were indistinguishable, and no charge nonuniformity was measured for any of the samples. Thus, the data supportthat changes in retention resulted from the different aspect ratios rather than from different surface chemistries. Interpretation of the retention data in terms of a collision efficiency (alpha) showed an increase with aspect ratio in both IS solutions, and for 1 mM the alpha increased from 0.011 (1:1) to 0.095 (2:1) to 0.26 (3:1). These results demonstrate for the first time the direct impact of particle shape on retention in porous media. Our findings have important implications for the transport of particles with high aspect ratios, such as rod-shaped bacteria, and for the modeling of such transport.

Inhibition of biohydrogen production by ammonia.

Ammonia inhibition of biohydrogen production was investigated in batch and continuous flow reactors with glucose as a substrate. In batch tests, biohydrogen production rate was highly dependent on pH and ammonia (defined as the sum of NH3 of NH4+ species) concentrations above 2 g N/L. At pH = 6.2, the maximum production decreased from 56 mL/h at 2 g N/L to 16 mL/h at 10 g N/L. At pH = 5.2, production decreased from 49 mL/h (2g N/L) to 7 mL/h (16 g N/L). Hydrogen yield remained relatively constant in batch tests, varying from 0.96 to 1.17 mol-H2/mol-glucose. In continuous flow tests, both hydrogen production rates and yields were adversely affected by ammonia. When the reactor (2.0 L) was first acclimated under batch conditions to a low nitrogen concentration (<0.8 g N/L), H2 production and yields under continuous flow mode conditions were 170 mL/h and 1.9 mol-H2/mol-glucose, but decreased with increased ammonia concentrations up to 7.8 g N/L to 105 mL/h and 1.1 mol-H2/mol-glucose. There was no hydrogen production under continuous flow conditions if the reactor was initially operated under batch flow conditions at ammonia concentrations above 0.8 g N/L. It is concluded that the hydrogen production is possible at high concentrations (up to 7.8 g N/L) of ammonia in continuous flow systems as long as the reactor is initially acclimated to a lower ammonia concentration (<0.8 g N/L).

Differences between chemisorbed and physisorbed biomolecules on particle deposition to hydrophobic surfaces.

This study examines differences between chemisorbed and physisorbed biomolecules on bacterial adhesion to both hydrophobic and hydrophilic surfaces that are biologically nonspecific. Bacteria-sized latex microspheres were used as a simplified model in order to study these factors that affect microbial adhesion. Two biomolecules (protein A, poly-D-lysine) were covalently bound to microspheres in order to study the effect of proteins on particle filtration rates in columns packed with glass beads. When poly-D-lysine or protein A was covalently bonded to the microspheres, sticking coefficients (a) for the microspheres increased by up to an order of magnitude as compared with uncoated latex microspheres. The glass packing beads were then made hydrophobic by covalently attaching silane groups with different carbon-chain lengths (0.2, 1.2, and 2.8 nm). Sticking coefficients forthe uncoated microspheres on these silanized packing beads (alpha = 0.15 at 1 mM ionic strength; 0.76 at 100 mM) were larger than those on uncoated glass packing beads (0.02 at 1 mM; 0.15 at 100 mM). In addition, adhesion increased with ionic strength on both hydrophobic and hydrophilic surfaces. Physical adsorption gave different results. When either dextran or protein A was physically adsorbed to both the microspheres and the column, no appreciable change in adhesion was observed. Covalently attaching protein A to the microspheres increased their hydrophobicity, but sticking coefficients were large regardless of the substrate hydrophobicity as a result of biomolecule-surface interactions. This study demonstrates that, at high ionic strength, covalently attached hydrophobic species give much higher sticking coefficients for particles than do physically adsorbed species.

Importance of molecular details in predicting bacterial adhesion to hydrophobic surfaces.

Electrostatic and hydrophobic forces are generally recognized as important in bacterial adhesion. Current continuum models for these forces often wrongly predict measurements of bacterial adhesion forces. The hypothesis tested here is that even qualitative guides to bacterial adhesion often require more than continuum information about hydrophobic forces; they require knowledge about molecular details of the bacteria and substrate surface. In this study, four different strains of bacteria were adsorbed to silica surfaces hydrophobized with alkylsilanes. The thickness of the lipopolysaccharide layers varied on the different bacteria, and the lengths of the alkylsilane molecules were varied from experiment to experiment. Bacterial adhesion was assessed using column experiments and atomic force microscopy (AFM) experiments. Results show that hydrophobized surfaces have higher bacterial sticking coefficients and stronger adhesion forces than bare silica surfaces, as expected. However, adhesion decreased as the solution Debye length became longer than the alkylsilane, perhaps since the silane molecules could not "reach" the bacterial surface. Similarly, those bacteria with a long o-antigen layer had decreased adhesion, perhaps since the silane molecules could not reach surface-bound proteins on the bacteria. This study reveals that macroscopic measurements such as contact angle are not able to fully describe bacterial adhesion; rather, additional details such as the molecular length are required to predict adhesion.