Steven W. Van Ginkel

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.

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. Shannons diversity index (H) and Simpsons 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.

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 H2 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.

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.

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.

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.

Impact of precipitation on the treatment of real ion-exchange brine using the H2-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.

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).

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.