Enriching acid rock drainage related microbial communities from surface-deposited oil sands tailings.

Little is known about the microbial communities native to surface-deposited pyritic oil sands tailings, an environment where acid rock drainage (ARD) could occur. The goal of this study was to enrich sulfur-oxidizing organisms from these tailings and determine whether different populations exist at pH levels 7, 4.5, and 2.5. Using growth-based methods provides model organisms for use in the future to predict potential activities and limitations of these organisms and to develop possible control methods. Thiosulfate-fed enrichment cultures were monitored for approximately 1 year. The results showed that the enrichments at pH 4.5 and 7 were established quicker than at pH 2.5. Different microbial community structures were found among the 3 pH environments. The sulfur-oxidizing microorganisms identified were most closely related to Halothiobacillus neapolitanus, Achromobacter spp., and Curtobacterium spp. While microorganisms related to Chitinophagaceae and Acidocella spp. were identified as the only possible iron-oxidizing and -reducing microbes. These results contribute to the general knowledge of the relatively understudied microbial communities that exist in pyritic oil sands tailings and indicate these communities may have a potential role in ARD generation, which may have implications for future tailings management.

The presence of nitrate dramatically changed the predominant microbial community in perchlorate degrading cultures under saline conditions.

BACKGROUND: Perchlorate contamination has been detected in both ground water and drinking water. An attractive treatment option is the use of ion-exchange to remove and concentrate perchlorate in brine. Biological treatment can subsequently remove the perchlorate from the brine. When nitrate is present, it will also be concentrated in the brine and must also be removed by biological treatment. The primary objective was to obtain an in-depth characterization of the microbial populations of two salt-tolerant cultures each of which is capable of metabolizing perchlorate. The cultures were derived from a single ancestral culture and have been maintained in the laboratory for more than 10 years. One culture was fed perchlorate only, while the other was fed both perchlorate and nitrate. RESULTS: A metagenomic characterization was performed using Illumina DNA sequencing technology, and the 16S rDNA of several pure strains isolated from the mixed cultures were sequenced. In the absence of nitrate, members of the Rhodobacteraceae constituted the prevailing taxonomic group. Second in abundance were the Rhodocyclaceae. In the nitrate fed culture, the Rhodobacteraceae are essentially absent. They are replaced by a major expansion of the Rhodocyclaceae and the emergence of the Alteromonadaceae as a significant community member. Gene sequences exhibiting significant homology to known perchlorate and nitrate reduction enzymes were found in both cultures. CONCLUSIONS: The structure of the two microbial ecosystems of interest has been established and some representative strains obtained in pure culture. The results illustrate that under favorable conditions a group of organisms can readily dominate an ecosystem and yet be effectively eliminated when their advantage is lost. Almost all known perchlorate-reducing organisms can also effectively reduce nitrate. This is certainly not the case for the Rhodobacteraceae that were found to dominate in the absence of nitrate, but effectively disappeared in its presence. This study is significant in that it reveals the existence of a novel group of organisms that play a role in the reduction of perchlorate under saline conditions. These Rhodobacteraceae especially, as well as other organisms present in these communities may be a promising source of unique salt-tolerant enzymes for perchlorate reduction.

Retroreflective imaging system for optical labeling and detection of microorganisms.

A retroreflective imaging system for imaging microscopic targets over macroscopic sampling areas is introduced. Detection of microorganism-bound retroreflector (RR) targets across millimeter-scale samples is implemented according to retroreflection directionality, collimation, and contrast design characteristics. Retroreflection directionality is considered for corner-cube (CC) and spherical geometries. Spherical-RRs improve directionality and reliability. Retroreflection collimation is considered for spherical-RRs. Retroreflective images for micro-CC-RRs and micro-spherical-RRs with varying refractive indices show optimal results for high refractive index BaTiO3 micro-spherical-RRs. A differential imaging technique improves retroreflection contrast by 35 dB. High refractive index micro-spherical-RRs and differential imaging, together, can detect microscopic RR targets across macroscopic areas.

Kinetics analysis of a salt-tolerant perchlorate-reducing bacterium: effects of sodium, magnesium, and nitrate.

Salt-tolerant perchlorate-reducing bacteria can be used to regenerate ion-exchange brines or resins exhausted with perchlorate. A salt-tolerant perchlorate-reducing Marinobacter vinifirmus strain P4B1 was recently purified. This study determined the effects of Na(+) and Mg(2+) concentrations on the perchlorate reduction rate of P4B1. The results showed that strain P4B1 could utilize perchlorate and grow in the presence of 1.8% to 10.2% NaCl. Lower NaCl concentrations allowed faster perchlorate reduction. The addition of Mg(2+) to the culture showed significant effects on perchlorate reduction when perchlorate was the sole electron acceptor. A molar Mg(2+)/Na(+) ratio of ∼0.11 optimized perchlorate degradation and cell growth. When perchlorate and nitrate were both present, nitrate reduction did not start significantly until perchlorate was below 100 mg/L. Tests with washed cell suspensions indicated that strain P4B1 had both perchlorate and nitrate reduction enzymes. When the culture was exposed to both perchlorate and nitrate, the nitrate reduction enzyme activity was low. The maximum specific substrate utilization rate (Vm) and the half saturation coefficient (KS) for P4B1 (30 g/L NaCl) determined in this study were 0.049 ± 0.003 mg ClO4(-)/mg VSS-h and 18 ± 4 mg ClO4(-)/L, respectively.

Synthesis and study of CuNiTiO3 as an ORR electrocatalyst to enhance microbial fuel cell efficiency.

Microbial fuel cells (MFCs) have the capability of simultaneous sewage treatment and electricity generation. Modifying the cathode electrode enhances their efficiency. In this study, NiTiO3 and CuNiTiO3 were synthesized for practical application as cathode catalysts in a dual-chamber MFC and the performance of the modified cathodes was evaluated against a bare graphite electrode. SEM images showed that the particle sizes were mostly in the range of 40-120 and 20-80 nm for NiTiO3 and CuNiTiO3, respectively. According to AFM results, CuNiTiO3 presented a higher surface roughness than NiTiO3. MFC using CuNiTiO3/G electrode with a reduction potential value of -0.27 V (vs. SCE) and a power density of 62.18 mW m-2 showed better oxygen reduction reaction (ORR) activity compared with NiTiO3/G and the bare graphite. MFC using CuNiTiO3 cathode also showed the highest values in terms of chemical oxygen demand (COD) removal (75%) and the calculated coulombic efficiency (CE, 10%). The results obtained in this study, introduce CuNiTiO3 as a promising electrocatalyst for further improvement of the cathodic reactions in MFC applications.

A review of anaerobic treatment of saline wastewater.

Large volumes of saline (> 2% w/v NaCl) wastewaters are discharged from many industries; e.g. seafood processing, textile dyeing, oil and gas production, tanneries and drinking water treatment processes. Although anaerobic treatment would be the most cost-effective and sustainable technology for the treatment of many of these saline wastewaters, the salinity is considered to be inhibitory to anaerobic biological treatment processes. The recent applications of salt-tolerant cultures for the treatment of wastewaters from seafood processing and ion-exchange processes suggest that biological systems can be used to treat salty wastewaters. Additionally, organisms capable of anaerobic degradation of contaminants in saline solutions have been observed in marine sediments and have been characterized during the last two decades. This manuscript provides a review of the recent research on anaerobic treatment of saline wastewater and bacterial consortia capable of the anaerobic degradation of pollutants in saline solutions, documenting that the biological treatment of saline wastewaters is promising.

Perchlorate and nitrate treatment by ion exchange integrated with biological brine treatment.

Groundwater contaminated with perchlorate and nitrate was treated in a pilot plant using a commercially available ion exchange (IX) resin. Regenerant brine concentrate from the IX process, containing high perchlorate and nitrate, was treated biologically and the treated brine was reused in IX resin regeneration. The nitrate concentration of the feed water determined the exhaustion lifetime (i.e., regeneration frequency) of the resin; and the regeneration condition was determined by the perchlorate elution profile from the exhausted resin. The biological brine treatment system, using a salt-tolerant perchlorate- and nitrate-reducing culture, was housed in a sequencing batch reactor (SBR). The biological process consistently reduced perchlorate and nitrate concentrations in the spent brine to below the treatment goals of 500 microg ClO4(-)/L and 0.5mg NO3(-)-N/L determined by equilibrium multicomponent IX modeling. During 20 cycles of regeneration, the system consistently treated the drinking water to below the MCL of nitrate (10 mgNO3(-)-N/L) and the California Department of Health Services (CDHS) notification level of perchlorate (i.e., 6 microg/L). A conceptual cost analysis of the IX process estimated that perchlorate and nitrate treatment using the IX process with biological brine treatment to be approximately 20% less expensive than using the conventional IX with brine disposal.

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.

Draft Genome Sequence of Marinobacter vinifirmus Type Strain FB1.

The gammaproteobacterium Marinobacter vinifirmus is associated with moderately saline environments and is often found in marine ecosystems. Here, we report the draft genome sequence of M. vinifirmus type strain FB1 (3.8 Mbp, 3,588 predicted genes). The presented sequence will improve our understanding of the taxonomy and evolution of the genus Marinobacter.

Mathematical modelling and reactor design for multi-cycle bioregeneration of nitrate exhausted ion exchange resin.

Nitrate contamination is one of the largest issues facing communities worldwide. One of the most common methods for nitrate removal from water is ion exchange using nitrate selective resin. Although these resins have a great capacity for nitrate removal, they are considered non regenerable. The sustainability of nitrate-contaminated water treatment processes can be achieved by regenerating the exhausted resin several times rather than replacing and incineration of exhausted resin. The use of multi-cycle exhaustion/bioregeneration of resin enclosed in a membrane has been shown to be an effective and innovative regeneration method. In this research, the mechanisms for bioregeneration of resin were studied and a mathematical model which incorporated physical desorption process with biological removal kinetics was developed. Regardless of the salt concentration of the solution, this specific resin is a pore-diffusion controlled process (XδD ¯CDr0(5+2α)<<1). Also, Thiele modulus was calculated to be between 4 and 12 depending on the temperature and salt concentration. High Thiele modulus (>3) shows that the bioregeneration process is controlled by reaction kinetics and is governed by biological removal of nitrate. The model was validated by comparison to experimental data; the average of R-squared values for cycle 1 to 5 of regeneration was 0.94 ± 0.06 which shows that the developed model predicted the experimental results very well. The model sensitivity for different parameters was evaluated and a model bioreactor design for bioregeneration of highly selective resins was also presented.

Draft Genome Sequence of Marinobacter sp. Strain P4B1, an Electrogenic Perchlorate-Reducing Strain Isolated from a Long-Term Mixed Enrichment Culture of Marine Bacteria.

The perchlorate-reducing strain Marinobacter sp. strain P4B1 was isolated from a long-term perchlorate-degrading enrichment culture seeded with marine sediment. The draft genome of Marinobacter sp. P4B1 is comprised of the bacterial chromosome (3.60 Mbp, G+C 58.51%, 3,269 predicted genes) and its associated plasmid pMARS01 (0.14 Mbp, G+C 52.95%, 165 predicted genes).

Effect of temperature & salt concentration on salt tolerant nitrate-perchlorate reducing bacteria: Nitrate degradation kinetics.

The sustainability of nitrate-contaminated water treatment using ion-exchange processes can be achieved by regenerating the exhausted resin several times. Our previous study shows that the use of multi-cycle bioregeneration of resin enclosed in membrane is an effective and innovative regeneration method. In this research, the effects of two independent factors (temperature and salt concentration) on the biological denitrification rate were studied. The results of this research along with the experimental results of the previous study on the effect of the same factors on nitrate desorption rate from the resin allow the optimization of the bioregeneration process. The results of nitrate denitrification rate study show that the biodegradation rate at different temperature and salt concentration is independent of the initial nitrate concentration. At each specific salt concentration, the nitrate removal rate increased with increasing temperature with the average value of 0.001110 ± 0.0000647 mg-nitrate/mg-VSS.h.°C. However, the effect of different salt concentrations was dependent on the temperature; there is a significant interaction between salt concentration and temperature; within each group of temperatures, the nitrate degradation rate decreased with increasing the salt concentration. The temperature affected the tolerance to salinity and culture was less tolerant to high concentration of salt at low temperature. Evidenced by the difference between the minimum and maximum nitrate degradation rate being greater at lower temperature. At 35 °C, a 32% reduction in the nitrate degradation rate was observed while at 12 °C this reduction was 69%. This is the first published study to examine the interaction of salt concentration and temperature during biological denitrification.

Sustainable nitrate-contaminated water treatment using multi cycle ion-exchange/bioregeneration of nitrate selective resin.

The sustainability of ion-exchange treatment processes using high capacity single use resins to remove nitrate from contaminated drinking water can be achieved by regenerating the exhausted resin and reusing it multiple times. In this study, multi cycle loading and bioregeneration of tributylamine strong base anion (SBA) exchange resin was studied. After each cycle of exhaustion, biological regeneration of the resin was performed using a salt-tolerant, nitrate-perchlorate-reducing culture for 48 h. The resin was enclosed in a membrane to avoid direct contact of the resin with the culture. The results show that the culture was capable of regenerating the resin and allowing the resin to be used in multiple cycles. The concentrations of nitrate in the samples reached a peak in first 0.5-1h after placing the resin in medium because of desorption of nitrate from resin with desorption rate of 0.099 ± 0.003 hr(-1). After this time, since microorganisms began to degrade the nitrate in the aqueous phase, the nitrate concentration was generally non-detectable after 10h. The average of calculated specific degradation rate of nitrate was -0.015 mg NO3(-)/mg VSS h. Applying 6 cycles of resin exhaustion/regeneration shows resin can be used for 4 cycles without a loss of capacity, after 6 cycles only 6% of the capacity was lost. This is the first published research to examine the direct regeneration of a resin enclosed in a membrane, to allow reuse without any disinfection or cleaning procedures.

Characterization of microbial populations in pilot-scale fluidized-bed reactors treating perchlorate- and nitrate-laden brine.

A salt-tolerant, perchlorate- and nitrate-reducing bacterial culture developed previously was used to inoculate two acetate-fed fluidized bed reactors (FBRs) which treated a 6% ion-exchange regenerant brine containing 500 +/- 84 mg-N/L nitrate and 4.6 +/- 0.6 mg/L perchlorate. The reactors were operated in series in continuous flow mode for 107 days after an acclimation period of 65 days. Pilot operation data suggest that complete denitrification was achieved after 70 days of operation, but significant perchlorate removal was not observed. Molecular analysis of the inoculum culture and biomass from the pilot plant samples using denaturing gradient gel electrophoresis (DGGE) and fluorescence in situ hybridization (FISH) revealed that the composition of the biomass in the pilot-plant was evolving with time in each FBR. The total number of Azoarcus/Denitromonas decreased in the first reactor with time and position in the bioreactor during acclimation and operation. FISH analysis clearly revealed that the number of Halomonas which was the dominant nitrate-reducing organism increased in the first reactor. This indicates a shift towards nitrate reduction which corresponds to the operation data. Both DGGE and FISH demonstrated that the Azoarcus/Denitromonas was still present in the second bioreactor, which indicated that the removal of nitrate in the first reactor was allowing the perchlorate-reducing organisms to establish themselves in the second reactor. The study also suggests that FISH was more effective for analysis of the composition of these cultures and it would be a better tool for the routine monitoring of cultures.

Biodegradation of synthetic base fluid surrogates in Gulf of Mexico sediments under simulated deep-sea conditions.

In this study, an anaerobic marine biodegradability test was adapted to study the fate of synthetic base fluid (SBF) surrogates, ethyl oleate and tetradecene, by deep-sea microorganisms. Sediment samples from hundreds of meters deep in the Gulf of Mexico were incubated at low temperatures (4 degrees C) and high hydrostatic pressure in steel vessels. Stimulation of indigenous microbial communities to SBF biodegradation was evident in the fact that the rate of removal of ethyl oleate was greater in sediments that had some previous exposure to SBF (first-order decay coefficient kof -0.22 +/- 0.02 week(-1)) compared to unexposed control sediments (first-order decay coefficient k of -0.11 +/- 0.02 week(-1)). When sulfate-linked tetradecene degradation occurred within the test period, the activity could also be modeled as a first-order decay following an initial lag phase, with an average decay coefficient of k = -0.05 +/- 0.01 week(-1). This study also revealed that the degradation of SBF surrogates by microorganisms collected from deep-sea sediments was not significantly effected by the hydrostatic incubation pressure.