Paul L. Bishop

Anaerobic/aerobic treatment of selected azo dyes in wastewater

Azo dyes represent the largest class of dyes in use today. Current environmental concern with these dyes revolves around the potential carcinogenic health risk presented by these dyes or their intermediate biodegradation products when exposed to microflora in the human digestive tract. These dyes may build up in the environment, since many wastewater treatment plants allow these dyes to pass through the system virtually untreated. The initial step in the degradation of these dyes is the cleavage of the Azo bond. This cleavage is often impossible under aerobic conditions, but has been readily demonstrated under anaerobic conditions. The focus of the study was to determine the feasibility of using an anaerobic fluidized-bed reactor to accomplish this cleavage. The effects of typical process variables such as hydraulic retention time (HRT), influent dye concentration levels, and degree of bed fluidization on removal efficiencies were also studied. The four dyes selected for this study were Acid-Orange 7, Acid-Orange 8, Acid-Orange 10, and Acid-Red 14. The effectiveness of using a bench-scale-activated sludge reactor as a sequenced second stage was also examined. Results indicate that nearly complete cleavage of the Azo bond is easily accomplished for each of the four dyes under hydraulic retention times of either 12 or 24 h. Initial results indicate, though, that aromatic amine by-products remain. The sequenced second stage was able to remove the remaining Chemical Oxygen Demand (COD) load to acceptable levels. Work is presently underway to determine the fate of the anaerobic by-products in the aerobic second stage.

Density, porosity, and pore structure of biofilms

Abstract The spatial distributions of biofilm properties have been investigated. Heterotrophic biofilms were cultured by using rotating drum biofilm reactors which were fed to a synthetic wastewater. The biofilm was first cut into 10–20 μm thick slices by use of a microtome, and then apportioned into samples representing 3 or 4 layers. By measuring total suspended solids, phospholipid concentrations and AR18 dye adsorption, and using these data with the theoretical model developed in this study, the densities, porosities, specific surface area, and mean pore radius of biofilms were determined. It was found that the densities (with units of mg-TS/cm3 total biofilm) in the bottom layers were 5–10 times higher than those in the top layers; the ratio of living cells to total biomass decreased from 72 to 91% in the top layers to 31–39% in the bottom layers; the porosities of biofilms changed from 84 to 93% in the top layers to 58–67% in the bottom layers. In contrast, the mean pore radius of biofilms decreased from approximately 1.7–2.7 μm in the top layers to 0.3–0.4 μm in the bottom layers. The results of this study are helpful in obtaining a much clearer physical description of biofilms.

Biodegradability of biofilm extracellular polymeric substances

This study discovered that biofilm extracellular polymeric substances (EPS) are biodegradable by their own producers and by other microorganisms when they are starved. The study was performed in a comparative fashion to examine the biodegradability of biofilm EPS by the microorganisms from the original biofilm (its own producers) and from activated sludge (other microorganisms). Four distinctive phases were observed during EPS biodegradation. In the first phase, instantaneous concentration increases of carbohydrate and protein in the test solutions were observed when EPS was added; in the second phase, easily biodegradable EPS from the added EPS was quickly utilized; in the third phase, microorganisms began to produce soluble EPS, using the minimally biodegradable EPS left from the previously added EPS; in the fourth phase, cells consumed the newly produced EPS and microbial activity gradually stopped. This study suggests that EPS can be used as a substrate, and that the EPS carbohydrate can be utilized faster than the EPS protein. The EPS utilization rates (including carbohydrate and protein) in the activated sludge suspension were greater than those in the biofilm suspension. It may take microorganisms longer to get acclimated to a new nutrient environment if they are in a starved state.

Effects of biofilm structure, microbial distributions and mass transport on biodegradation processes

The influence of biofilm structure on transport and transformation processes in biofilms has been investigated microscopically using microelectrodes, a micro-slicing procedure and various chemical and microbiological tests. The study demonstrates that the biofilm structure is highly stratified, characterized by an increase of biofilm density, a decrease of metabolically active biomass, and a decrease of porosity with biofilm depth. Both the effective diffusivity for dissolved oxygen and the effectiveness factor decrease with biofilm depth. Competition for substrate and space in biofilms results in this stratified structure, which is also affected by biofilm thickness. The study reveals that there are different trends for the density increase and the decreases of porosity, microbial activity and DO effective diffusivity with biofilm depth for different biofilm thicknesses. The results of this study are helpful in obtaining a clearer physical description of biofilms, and help to bridge the gap between the mathematical modelling and external-phenomenon observation of biofilm systems.

Competition for substrate and space in biofilms

Competition for substrate and space in biofilms was studied using a microelectrode technique and a micro-slicing technique. Three different kinds of biofilms were cultured using laboratory-scale, rotating drum biofilm reactors fed with synthetic wastewater. The measured concentration profiles provide direct experimental evidence of the competition in multispecies biofilms for substrates. Increases in organic loading or ammonium-nitrogen loading cause more consumption of oxygen, which results in competition for oxygen between heterotrophs and nitrifiers. Even in a pure nitrification system, heterotrophs, supported by soluble microbial products or metabolic products, could exist in the nitrification biofilm. Nitrifiers, however, have difficulty existing in the heterotrophic biofilms, and their populations were always 4 or 5 orders lower than those ofheterotrophs. It was found that the value of criterion for transition between oxygen and ammonium in nitrifying biofilms (S N /S DO ) was between 0.77 and 1.2, and the value decreased with an increase ofglucose loading. The competition for substrate in biofilms resulted in a stratified structure with nonuniform spatial distributions of biofilm properties, such as density, porosity, and effective diffusivity. This stratified structure in turn affects the substrate transfer and substrate competition within the biofilm. It was found that a certain biofilm system may not have only one penetration depth, corresponding to the critical thickness, for the whole range of biofilm thicknesses.

Evaluation of tortuosity factors and effective diffusivities in biofilms

Abstract A cylindrical model (CM) and a random porous cluster model (RPCM) have been developed in this paper to evaluate tortuosities, tortuosity factors, and effective diffusivities within biofilms, based on measurable biofilm properties. The tortuosity factor and the ratio of effective diffusivities to diffusivities in bulk solution ( D e /D b ) of the biofilm can be evaluated from biofilm porosity data by employing the CM presented in this paper. Both tortuosities and effective diffusivities depend on the densities and porosities of biofilms. For biofilms with porosities of 0.84–0.93 in the top layers and 0.58–0.67 in the bottom layer, it was found that tortuosity factors increase from 1.15 to approximately 1.6, while ratios of ( D e /D b ) decrease from 68–81% in the top layer to 38–45% in the bottom layer, based on the CM. The spatial distributions of biofilm properties affect the effectiveness factor profiles directly. Using incorrect ratios of ( D e /D b ) would result in false modeling results.

Characterization and evaluation of aerobic granules in sequencing batch reactor

In order to investigate the aerobic granules cultured under alternating aerobic and anoxic conditions, a sequencing batch reactor (SBR) was operated without the presence of a carrier material. Nitrification and denitrification occurred alternately in the SBR operation, with an increased nitrification efficiency of up to 97% and a high chemical oxygen demand (COD) removal efficiency of up to 95%. It was observed that physical characteristics of granule play an important role in the performance of the SBR process. Light microscopy was used to observe the time dependent development of the granules in the SBR. Based on the microscopic observations, some floc-like sludges remained in the form of a mixture with granules for 30 days of operation. Even though various granule sizes had been formed in the reactor after 50 days, the granule sizes were primarily from 1 +/- 0.35 to 1.3 +/- 0.45 mm, rarely exceeding 2 mm. The granules were analyzed by a combination of microelectrodes and fluorescent in situ hybridization (FISH), which provides more detailed information on what happens inside the granules. Based on their results, ammonia oxidizing bacteria (AOB) existed primarily in the upper and middle layers of the granule. Assuming a first-order reaction for nitrification, most of the nitrification is likely to occur from the surface to 300 microm into the granular thickness.

Degradation of acid orange 7 in an aerobic biofilm.

A stable microbial biofilm community capable of completely mineralizing the azo dye acid orange 7 (AO7) was established in a laboratory scale rotating drum bioreactor (RDBR) using waste liquor from a sewage treatment plant. A broad range of environmental conditions including pH (5.8-8.2), nitrification (0.0-4.0 mM nitrite), and aeration (0.2-6.2 mg O2 l(-1)) were evaluated for their effects on the biodegradation of AO7. Furthermore the biofilm maintained its biodegradative ability for over a year while the effects of these environmental conditions were evaluated. Reduction of the azo bond followed by degradation of the resulting aromatic amine appears to be the mechanism by which this dye is biodegraded. Complete loss of color, sulfanilic acid, and chemical oxygen demand (COD) indicate that AO7 is mineralized. To our knowledge this is the first reported occurrence of a sulfonated phenylazonaphthol dye being completely mineralized under aerobic conditions. Two bacterial strains (ICX and SAD4i) originally isolated from the RDBR were able to mineralize, in co-culture, up to 90% of added AO7. During mineralization of AO7, strain ICX reduces the azo bond under aerobic conditions and consumes the resulting cleavage product 1-amino-2-naphthol. Strain SAD4i consumes the other cleavage product, sulfanilic acid. The ability of the RDBR biofilm to aerobically mineralize an azo dye without exogenous carbon and nitrogen sources suggests that this approach could be used to remediate industrial wastewater contaminated with spent dye.

A microelectrode study of redox potential change in biofilms

In this study, we examined the stratification of microbial processes and the associated redox potential changes in biofilms using microelectrode techniques. Two types of biofilms, each with a different combination of microbial processes, were examined. The first type carried aerobic oxidation and sulfate reduction, while the second one provided aerobic oxidation and nitrification. The microelectrodes used were oxygen, sulfide, ammonium, pH and redox potential microelectrodes. The results of this study provide the following new experimental evidence: (1) The aerobic/sulfate-reducing biofilm had a clearly stratified structure with depth. In this biofilm, aerobic oxidation took place only in a shallow layer near the surface and sulfate reduction occurred in the deeper anoxic zone. The boundary between these two processes was well defined. (2) The aerobic/nitrifying biofilm also had a stratified structure with depth. In this biofilm, though aerobic oxidation took place throughout the biofilm depth, more nitrification occurred in the deeper section of the biofilm. The boundary between these two processes, however, was less well defined. (3) Redox potential could be an indicator for the existence of certain microbial processes in biofilms. The redox potential profile changes were correlated to shifts of microbial processes in both types of biofilms. The redox potential profiles in these biofilms can be used to elucidate the stratification of microbial processes in the biofilms.

Simultaneous anaerobic and aerobic degradation of the sulfonated azo dye Mordant Yellow 3 by immobilized cells from a naphthalenesulfonate-degrading mixed culture

Abstract The naphthalenesulfonate-oxidizing bacterium Sphingomonas sp. BN6 was immobilized in calcium alginate. These beads were incubated under aerobic conditions in a medium with the sulfonated azo dye, Mordant Yellow 3 (MY3), and glucose. The immobilized cells converted MY3, but only a marginal turnover of the dye was found under these conditions with freely suspended cells of Sphingomonas sp. BN6. Under anaerobic conditions, suspended cells of Sphingomonas sp. BN6 reductively cleaved the azo bond of MY3 to 6-aminonaphthalene-2-sulfonate (6A2NS) and 5-aminosalicylate. The turnover of MY3 by the immobilized cells under aerobic conditions resulted in the formation of more than equimolar amounts of 5-aminosalicylate, but almost no (6A2NS) was detected. Cells of Sphingomonas sp. BN6 aerobically oxidize 6A2NS to 5-aminosalicylate. It was therefore concluded that the cells in the anaerobic center of the alginate beads reduced MY3 to 6A2NS and 5-aminosalicylate and that 6A2NS was oxidized to 5-aminosalicylate by those cells that were immobilized in the outer aerobic zones of the alginate beads. The presence of oxygen gradients within the alginate beads was verified by using oxygen micro-electrodes. A coimmobilisate of Sphingomonas sp. BN6 with a 5-aminosalicylate degrading bacterium completely degraded MY3. The immobilized cells also converted the sulfonated azo dyes Amaranth and Acid Red␣1.