Electron flow of biological H2 production by sludge under simple thermal treatment: Kinetic study.

Mixed culture sludge has been widely used as a microbial consortium for biohydrogen production. Simple thermal treatment of sludge is usually required in order to eliminate any H2-consuming bacteria that would reduce H2 production. In this study, thermal treatment of sludge was carried out at various temperatures. Electron flow model was then applied in order to assess community structure in the sludge upon thermal treatment for biohydrogen production. Results show that the dominant electron sink was acetate (150-217 e- meq/mol glucose). The electron equivalent (e- eq) balances were within 0.8-18% for all experiments. Treatment at 100 °C attained the highest H2 yield of 3.44 mol H2/mol glucose from the stoichiometric reaction. As the treatment temperature increased from 80 to 100 °C, the computed acetyl-CoA and reduced form of ferredoxin (Fdred) concentrations increased from 13.01 to 17.34 e- eq (1.63-2.17 mol) and 1.34 to 4.18 e- eq (0.67-2.09 mol), respectively. The NADH2 balance error varied from 3 to 10% and the term e-(Fd↔NADH2) (m) in the NADH2 balance was NADH2 consumption (m = -1). The H2 production was mainly via the Fd:hydrogenase system and this is supported with a good NADH2 balance. Using the modified Gompertz model, the highest maximum H2 production potential was 1194 mL whereas the maximum rate of H2 production was 357 mL/h recorded at 100 °C of treatment.

Determining the Mechanism for Low Sludge Yields in the Cannibal Solids Reduction System.

Extracellular polymeric substances (EPS) and solids concentrations in samples from the Interchange Bioreactor (IBR), and return activated sludge (RAS) from Cannibal facilities having low and high sludge yields, were analyzed to understand the mechanisms behind low sludge production. Low sludge yields correlated to more EPS degradation, higher concentrations of iron, and reducing conditions in the IBR. In the low yield facilities, iron was reduced when the RAS passed through the anaerobic and reducing conditions of the IBR, and this led to more EPS solubilization and degradation. This "futile cycle" of EPS production and degradation appears to may have been most responsible for reducing sludge yields.

Toxicity assessment using different bioassays and microbial biosensors.

Toxicity assessment of water streams, wastewater, and contaminated sediments, is a very important part of environmental pollution monitoring. Evaluation of biological effects using a rapid, sensitive and cost effective method can indicate specific information on ecotoxicity assessment. Recently, different biological assays for toxicity assessment based on higher and lower organisms such as fish, invertebrates, plants and algal cells, and microbial bioassays have been used. This review focuses on microbial biosensors as an analytical device for environmental, food, and biomedical applications. Different techniques which are commonly used in microbial biosensing include amperometry, potentiometry, conductometry, voltammetry, microbial fuel cells, fluorescence, bioluminescence, and colorimetry. Examples of the use of different microbial biosensors in assessing a variety of environments are summarized.

The Selective Use of Hypochlorite to Prevent Pond Crashes for Algae-Biofuel Production.

Although algae-biofuels have many advantages including high areal productivity, algae can be preyed upon by amoebas, protozoans, ciliates, and rotifers, particularly in open pond systems. Thus, these higher organisms need to be controlled. In this study, Chlorella kessleri was used as the algal culture and Brachionus calyciflorus as the source of predation. The effect of sodium hypochlorite (bleach) was tested with the goal of totally inhibiting the rotifer while causing minor inhibition to the alga. The 24-hr LC(50) for B. calyciflorus in spring water was 0.198 mg Cl/L while the 24-hr LC(50) for C. kessleri was 0.321 mg Cl/L. However, chlorine dissipates rapidly as the algae serves as reductant. Results showed a chlorine dosage between 0.45 to 0.6 mg Cl/L and a dosing interval of two hours created the necessary chlorine concentrations to inhibit predation while letting the algae grow; thus giving algae farmers a tool to prevent pond crashes.

Use of Copper to Selectively Inhibit Brachionus calyciflorus (Predator) Growth in Chlorella kessleri (Prey) Mass Cultures for Algae Biodiesel Production.

A single Brachionus rotifer can consume thousands of algae cells per hour causing an algae pond to crash within days of infection. Thus, there is a great need to reduce rotifers in order for algal biofuel production to become reality. Copper can selectively inhibit rotifers in algae ponds, thereby protecting the algae crop. Differential toxicity tests were conducted to compare the copper sensitivity of a model rotifer-B. calyciflorus and an alga, C. kessleri. The rotifer LC50 was <0.1 ppm while the alga was not affected up to 5 ppm Cu(II). The low pH of the rotifer stomach may make it more sensitive to copper. However, when these cultures were combined, a copper concentration of 1.5 ppm was needed to inhibit the rotifer as the alga bound the copper, decreasing its bioavailability. Copper (X ppm) had no effect on downstream fatty acid methyl ester extraction.

Effect of centrifugation on water recycling and algal growth to enable algae biodiesel production.

The latest research shows that algal biofuels, at the production levels mandated in the Energy Independence and Security Act of 2007, will place significant demands on water and compete with agriculture meant for food production. Thus, there is a great need to recycle water while producing algal biofuels. This study shows that when using a synthetic medium, soluble algal products, bacteria, and other inhibitors can be removed by centrifugation and enable water recycling. Average water recovery reached 84% and water could be recycled at least 10 times without reducing algal growth.

Effect of trace tetracycline concentrations on the structure of a microbial community and the development of tetracycline resistance genes in sequencing batch reactors.

The objective of this study was to investigate effects of different concentrations of tetracycline (TC) on the microbial community and development of tetracycline resistance genes (TRGs) of sequencing batch reactors (SBRs). Polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE) analysis of 16S rRNA and real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) were used to detect the structural changes of the microbial community and the variations of eight TC resistance genes tet(A), tet(B), tet(C), tet(E), tet(M), tet(O), tet(S) and tet(X), respectively. The results indicated that, trace TC could substantially change the structure of the microbial community. Bacteria which could not adapt to environment with TC were gradually replaced by those adapting to tetracycline. Shannon's diversity index (H) and Simpson's index (D) reached maximum values when the concentration of TC was 1 µg L(-1). The resistance genes in the activated sludge proliferated under the pressure of trace TC.

Effect of organics and alkalinity on the sulfur oxidizing bacteria (SOB) biosensor.

The environmental risk assessment of toxic chemicals in stream water requires the use of a low cost standardized toxicity bioassay. Here, a biosensor for detection of toxic chemicals in stream water was studied using sulfur oxidizing bacteria (SOB) in continuous mode. The biosensor depends on the ability of SOB to oxidize sulfur particles under aerobic conditions to produce sulfuric acid. The reaction results in an increase in electrical conductivity (EC) and a decrease in pH. The biosensor is based on the inhibition of SOB in the presence of toxic chemicals by measuring changes in EC and pH. We found that the SOB biosensor can detect Cr(6+)at a low concentration (50 ppb) which is lower than many whole-cell biosensors. The effect of organic material in real stream water on SOB activity was studied. Due to the presence of mixotrophic SOB, we found that the presence of organic matter increases SOB activity which decreases the biosensor start up period. Low alkalinity (22 mg L(-1) CaCO(3)) increased effluent EC and decreased effluent pH which is optimal for biosensor operation. While at high alkalinity (820 mg L(-1) CaCO(3), the activity of SOB little decreased. We found that system can detect 50 ppb of Cr(6+) at low alkalinity (22 mg L(-1) CaCO(3)) in few hours while, complete inhibition was observed after 35 h of operation at high alkalinity (820 mg L(-1) CaCO(3)).

Kinetic study of slaughterhouse wastewater treatment by electrocoagulation using Fe electrodes.

In this study, treatment of slaughterhouse wastewater by electrocoagulation was investigated in batch system using Fe electrodes. The effect of various variables such as electrode number, current density and operating time was tested. Pollutant removal efficiency increased with increasing electrode number and operating time. The biochemical oxygen demand (BOD(5))(,) chemical oxygen demand (COD), total suspended solid (TSS), and total nitrogen (TN) removal efficiencies using eight electrodes at a contact time of 50 min and a current density of 10 A/m(2) were 66, 62, 60, and 56%, respectively. Higher electrode numbers will allow shorter operating times to achieve certain removal efficiencies. Also, removal efficiencies increased by increasing the current density; the highest removal efficiencies of BOD(5,) COD, TSS, and TN at a contact time of 50 min and a current density of 25 A/m(2) were 97, 93, 81, and 84%, respectively. The results also show that the reactor pH varies directly with the current density; at 25 A/m(2), the reactor pH increased from an initial value of 7.1 to 7.7 after 50 min. The experimental results showed that the kinetics of BOD(5), COD, TSS and TN removal could be fitted adequately using a first order kinetic model (higher R(2)).

Detection of Cr6+ by the sulfur oxidizing bacteria biosensor: effect of different physical factors.

A biosensor based on sulfur-oxidizing bacteria (SOB) for detection of toxic chemicals in water was developed. SOB are acidophilic microorganisms that get their energy through the oxidation of reduced sulfur compounds in the presence of oxygen to produce sulfuric acid. The reaction results in an increase in electrical conductivity (EC) and a decrease in pH. The bioassay is based on the inhibition of SOB in the presence of toxic chemicals by measuring changes in EC and pH. The effect of different physical factors such as HRT, inorganic sulfur (S°) particle size, and temperature on detection of Cr(6+) was studied. The detection of Cr(6+) (50 ppb) was improved by decreasing the hydraulic retention time (HRT) from 30 to 10 min and increasing S° particle size from 1 to 4.75 mm. Detection time was shorter at 30 °C compared to 45 °C and the SOB were active over a wide range of temperatures with a maximum temperature for growth at 45 °C. This novel biosensor is simple, highly sensitive to low Cr(6+) concentrations (50 ppb), and also minimizes detection time. The present findings can be applied to the proper continuous screening of water ecosystem toxicity.

Effect of NaCl on nitrate removal from ion-exchange spent brine in the membrane biofilm reactor (MBfR).

The H(2)-based membrane biofilm reactor was used to remove nitrate from synthetic ion-exchange brine at NaCl concentrations from ∼3 to 30 g/L. NaCl concentrations below 20 g/L did not affect the nitrate removal flux as long as potassium was available to generate osmotic tolerance for high sodium, the H(2) pressure was adequate, and membrane fouling was eliminated. Operating pHs of 7-8 and periodic citric acid washes controlled membrane fouling and enabled reactor operation for 650 days. At 30 psig H(2) and high nitrate loading rates of 15 to 80 g/m(2) d, nitrate removal fluxes ranged from 2.5 to ∼6 g/m(2) d, which are the highest fluxes observed when treating 30 g/L IX brine. However, percent removals were low, and the H(2) pressure probably limited the removal flux.

Effect of pH on nitrate and selenate reduction in flue gas desulfurization brine using the H2-based membrane biofilm reactor (MBfR).

Increased tightening of air regulations is leading more electric utilities to install flue gas desulfurization (FGD) systems. These systems produce brine containing high concentrations of nitrate, nitrite, and selenate which must be removed before discharge. The H2-based membrane biofilm reactor (MBfR) was shown to consistently remove nitrate, nitrite, and selenate at high efficiencies. The maximum selenate removal flux reached 362 mgSe m(-2)d(-1) and was higher than that observed in earlier research, which shows continual improvement of the biofilm for selenate reduction. A low pH of 6.8 inhibited precipitation when treating actual FGD brine, yet did not inhibit removal. SO4(2-) was not removed and therefore did not compete with nitrate, nitrite, and selenate reduction for the available H2.

The removal of selenate to low ppb levels from flue gas desulfurization brine using the H2-based membrane biofilm reactor (MBfR).

The H(2)-based membrane biofilm reactor (MBfR) was shown to consistently remove nitrate, nitrite, and selenate at high efficiencies from flue-gas desulfurization brine. Selenate was removed to <50 ppb which is the National Pollutant Discharge Elimination System (NPDES) criteria for the brine to be released into the environment. When selenate was removed to <50 ppb, nitrate and nitrite were still present in the mg/L range which suggests that selenate is able to be secondarily reduced to low levels when nitrate and nitrite serve as the main electron acceptors for bacterial growth. SO(4)(2-) was not removed and therefore did not compete with nitrate and selenate reduction for the available H(2).

Impact of precipitation on the treatment of real ion-exchange brine using the H(2)-based membrane biofilm reactor.

The H(2)-based membrane biofilm reactor (MBfR) was used to remove nitrate and perchlorate from real ion-exchange brine at two different salinities (30- and 50-g/L NaCl). Base production from nitrate reduction to N(2) gas caused the pH to increase, and this exacerbated precipitation of calcium and magnesium carbonates onto the MBfR fibers. The precipitates lowered the H(2) flux to the biofilm and caused a deterioration of denitrification performance that could be reversed by mild citric-acid washing. The addition of acid seems to be the only mechanism to avoid serious precipitation, membrane fouling, and non-optimal pH for denitrification.

Detecting oxidized contaminants in water using sulfur-oxidizing bacteria.

For the rapid and reliable detection of oxidized contaminants (i.e., nitrite, nitrate, perchlorate, dichromate) in water, a novel toxicity detection methodology based on sulfur-oxidizing bacteria (SOB) has been developed. The methodology exploits the ability of SOB to oxidize elemental sulfur to sulfuric acid in the presence of oxygen. The reaction results in an increase in electrical conductivity (EC) and a decrease in pH. When oxidized contaminants were added to the system, the effluent EC decreased and the pH increased due to the inhibition of the SOB. We found that the system can detect these contaminants in the 5-50 ppb range (in the case of NO(3)(-), 10 ppm was detected), which is lower than many whole-cell biosensors to date. At low pH, the oxidized contaminants are mostly in their acid or nonpolar, protonated form which act as uncouplers and make the SOB biosensor more sensitive than other whole-cell biosensors which operate at higher pH values where the contaminants exist as dissociated anions. The SOB biosensor can detect toxicity on the order of minutes to hours which can serve as an early warning so as to not pollute the environment and affect public health.

Detecting endocrine disrupting compounds in water using sulfur-oxidizing bacteria.

For the rapid and reliable detection of endocrine disrupting compounds in water, a novel toxicity detection methodology based on sulfur-oxidizing bacteria (SOB) has been developed. The methodology exploits the ability of SOB to oxidize elemental sulfur to sulfuric acid in the presence of oxygen. The reaction results in an increase in electrical conductivity (EC) and a decrease in pH. When endocrine disrupting compounds were added to the system, the effluent EC decreased and the pH increased due to the inhibition of the SOB. We found that the system can detect these chemicals in the 50-200 ppb range, which is lower than many whole-cell biosensors to date. The SOB biosensor can detect toxicity on the order of min to h which can serve as an early warning so as to not pollute the environment and affect public health.

Isolation and characterization of Acidithiobacillus caldus from a sulfur-oxidizing bacterial biosensor and its role in detection of toxic chemicals.

A novel toxicity detection methodology based on sulfur-oxidizing bacteria (SOB) has been developed for the rapid and reliable detection of toxic chemicals in water. The methodology exploits the ability of SOB to oxidize sulfur particles in the presence of oxygen to produce sulfuric acid. The reaction results in an increase in electrical conductivity (EC) and a decrease in pH. The assay is based on the inhibition of SOB in the presence of toxic chemicals by measuring changes in EC and pH. We found that SOB biosensor can detect toxic chemicals, such as heavy metals and CN-, in the 5-2000ppb range. One bacterium was isolated from an SOB biosensor and the 16S rRNA gene of the bacterial strain has 99% and 96% sequence similarity to Acidithiobacillus sp. ORCS6 and Acidithiobacillus caldus DSM 8584, respectively. The isolate was identified as A. caldus SMK. The SOB biosensor is ideally suited for monitoring toxic chemicals in water having the advantages of high sensitivity and quick detection.

Microbial community structure during nitrate and perchlorate reduction in ion-exchange brine using the hydrogen-based membrane biofilm reactor (MBfR).

Detoxification of perchlorate by microbial communities under denitrifying conditions has been recently reported, although the identity of the mixed populations involved in perchlorate reduction is not well understood. In order to address this, the bacterial diversity of membrane biofilm reactors (MBfR) set up under autotrophic denitrifying and perchlorate-reducing conditions were examined by analyses of the 16S rRNA gene sequences of clone libraries. Inocula from diverse locations were tested for their ability to reduce nitrate and perchlorate in synthetic ion exchange spent brine (45g/l NaCl) using H(2)-based MBfRs. Phylogenetic analysis of 16S rRNA gene sequences showed that proteobacterial species dominated the biofilm communities, particularly nitrate-reducing gamma-proteobacteria. Even though the inocula to the MBfRs came from different sources, clones closely related to Marinobacter hydrocarbonoclasticus represented 53% of all clones in the MBfR biofilms. The clone libraries contained no known perchlorate-reducing bacteria, which suggest that denitrifiers carried out perchlorate reduction, probably by secondary-utilization.

Bacterial biofilm-community selection during autohydrogenotrophic reduction of nitrate and perchlorate in ion-exchange brine.

Three hydrogen-based membrane biofilm reactors (MBfR) biologically reduced nitrate and perchlorate in a synthetic ion-exchange (IX) brine. Inocula from different natural saline environments were employed to initiate the three MBfRs. Under high-salinity (3%) conditions, bioreduction of nitrate and perchlorate occurred simultaneously, and the three MBfRs from the different inocula exhibited similar removal fluxes for nitrate and perchlorate. Clone libraries were generated from samples of the biofilms in the three MBfRs and compared to those of their inocula. When H(2) was the sole exogenous electron donor under high-salinity conditions, MBfR-driven community shifts were observed with a similar pattern regardless of inoculum. The following 16S rRNA gene phylogenetic analysis showed the presence of novel perchlorate-reducing bacteria in the salt-tolerant mBfR communities. These findings suggest that autohydrogenotrophic and high-salinity conditions provided strong selective pressure for novel perchlorate-reducing populations in the mBfRs.

Kinetics of nitrate and perchlorate reduction in ion-exchange brine using the membrane biofilm reactor (MBfR).

Several sources of bacterial inocula were tested for their ability to reduce nitrate and perchlorate in synthetic ion-exchange spent brine (30-45 g/L) using a hydrogen-based membrane biofilm reactor (MBfR). Nitrate and perchlorate removal fluxes reached as high as 5.4 g Nm(-2)d(-1) and 5.0 g ClO(4)m(-2)d(-1), respectively, and these values are similar to values obtained with freshwater MBfRs. Nitrate and perchlorate removal fluxes decreased with increasing salinity. The nitrate fluxes were roughly first order in H(2) pressure, but roughly zero-order with nitrate concentration. Perchlorate reduction rates were higher with lower nitrate loadings, compared to high nitrate loadings; this is a sign of competition for H(2). Nitrate and perchlorate reduction rates depended strongly on the inoculum. An inoculum that was well acclimated (years) to nitrate and perchlorate gave markedly faster removal kinetics than cultures that were acclimated for only a few months. These results underscore that the most successful MBfR bioreduction of nitrate and perchlorate in ion-exchange brine demands a well-acclimated inoculum and sufficient hydrogen availability.