The synergistic interaction between sulfate-reducing bacteria and pyrogenic carbonaceous matter in DDT decay.

Although 1,1,1-trichloro-2,2-di(4-chlorophenyl)ethane (DDT) was banned in the United States in 1972, it is still often detected in sediments where pyrogenic carbonaceous matter (PCM) and sulfate-reducing bacteria (SRB) co-exist. In this study, we found that 70.2 ±â€¯0.2% of DDT disappeared in the presence of SRB and graphite powder, a model PCM, after 21 days at pH 7. Our results suggest that the observed DDT decay was due to the reaction between graphite powder and the reduced sulfur species that were produced by SRB. No biofilm formation was observed on the surface of graphite powder. Rather, the activity of SRB was inhibited by the presence of graphite powder. To understand the involvement of PCM in DDT decay, electrochemical cells and batch reactor experiments with sulfur-pretreated PCM as well as direct electrochemical reduction by a potentiostat were employed. Our results suggest that polysulfide, sulfide, sulfite, and thiosulfate could all react with PCM, forming surface-bound intermediates that subsequently led to DDT decay. The reactivity of reduced sulfur species was the highest for polysulfide, followed by sulfide, sulfite, and thiosulfate.

Bacteria inactivation at a sub-stoichiometric titanium dioxide reactive electrochemical membrane.

This study investigated the use of a sub-stoichiometric TiO2 reactive electrochemical membrane (REM) for the inactivation of a model Escherichia coli (E. coli) pathogen in chloride-free solutions. The filtration system was operated in dead-end, outside-in filtration model, using the REM as anode and a stainless steel mesh as cathode. A 1-log removal of E. coli was achieved when the electrochemical cell was operated at the open circuit potential, due to a simple bacteria-sieving mechanism. At applied cell potentials of 1.3 and 3.5V neither live nor dead E. coli cells were detected in the permeate stream (detection limit of 1.0 cell mL(-1)), which was attributed to enhanced electrostatic bacteria adsorption at the REM anode. Bacteria inactivation in the retentate solution increased as a function of the applied cell potential, which was attributed to transport of E. coli to the REM and stainless steel cathode surfaces, and direct contact with the local acidic and alkaline environment produced by water oxidation at the anode and cathode, respectively. Clear evidence for an E. coli inactivation mechanism mediated by either direct or indirect oxidation was not found. The low energy requirement of the process (2.0-88Whm(-3)) makes the REM an attractive method for potable water disinfection.

Comparison of Overall Resource Consumption of Biosolids Management System Processes Using Exergetic Life Cycle Assessment.

This study focused on the evaluation of biosolids management systems (BMS) from a natural resource consumption point of view. Additionally, the environmental impact of the facilities was benchmarked using Life Cycle Assessment (LCA) to provide a comprehensive assessment. This is the first study to apply a Cumulative Exergy Extraction from the Natural Environment (CEENE) method for an in-depth resource use assessment of BMS where two full-scale BMS and seven system variations were analyzed. CEENE allows better system evaluation and understanding of how much benefit is achievable from the products generated by BMS, which have valorization potential. LCA results showed that environmental burden is mostly from the intense electricity consumption. The CEENE analysis further revealed that the environmental burden is due to the high consumption of fossil and nuclear-based natural resources. Using Cumulative Degree of Perfection, higher resource-use efficiency, 53%, was observed in the PTA-2 where alkaline stabilization rather than anaerobic digestion is employed. However, an anaerobic digestion process is favorable over alkaline stabilization, with 35% lower overall natural resource use. The most significant reduction of the resource footprint occurred when the output biogas was valorized in a combined heat and power system.

Is aceticlastic methanogen composition in full-scale anaerobic processes related to acetate utilization capacity?

In this study, biomass samples were obtained from six municipal and nine industrial full-scale anaerobic processes to investigate whether the aceticlastic methanogen population composition is related to acetate utilization capacity and the nature of the wastewater treated, i.e. municipal sludge or industrial wastewater. Batch serum bottle tests were used to determine the specific acetate utilization rate (AUR), and a quantitative real-time polymerase chain reaction protocol was used to enumerate the acetate-utilizing Methanosaeta and Methanosarcina populations in the biomass samples. Methanosaeta was the dominant aceticlastic methanogen in all samples, except for one industrial wastewater-treating anaerobic process. However, Methanosarcina density in industrial biomass samples was higher than the Methanosarcina density in the municipal samples. The average AUR values of municipal and industrial wastewater treatment plant biomass samples were 10.49 and 10.65 mg CH3COO(-)/log(aceticlastic methanogen gene copy).d, respectively. One-way ANOVA test and principle component analysis showed that the acetate utilization capacities and aceticlastic methanogen community composition did not show statistically significant correlation among the municipal digesters and industrial wastewater-treating processes investigated.

The effect of managing nutrients in the performance of anaerobic digesters of municipal wastewater treatment plants.

Is it possible to create conditions in the anaerobic digesters to control nutrients without changing the performance of a reactor? This study investigates an answer for this question. To this purpose, anaerobic reactors are operated at high concentrations of Mg(2+) ion to harvest the nutrient ions (NH4 (+) and PO4 (3-)) in the form of struvite, that is, magnesium ammonium phosphate. The effects of this modification on the anaerobic digestion of sewage sludge were investigated in terms of chemical oxygen demand (COD) removal and cumulative CH4 production as well as the changes in the biological diversity. The results showed that approximately 50 % of the nutrients (NH4 (+) and PO4 (3-)) were removed regardless of the method adopted for the addition of Mg(2+) ion, slug or daily dosing. The numbers of Methanosaeta and Methanosarcina in the samples withdrawn prior to and after the addition of Mg(2+) did not show significant difference according to the results obtained from qPCR analyses. The research results showed that the addition of Mg(2+) into the anaerobic digesters in municipal wastewater treatment facilities may help to remove the nutrients from the effluent while recovering in their solid forms.

Selective quantification of viable Escherichia coli bacteria in biosolids by quantitative PCR with propidium monoazide modification.

Quantitative differentiation of live cells in biosolids samples, without the use of culturing-based approaches, is highly critical from a public health risk perspective, as recent studies have shown significant regrowth and reactivation of indicator organisms. Persistence of DNA in the environment after cell death in the range of days to weeks limits the application of DNA-based approaches as a measure of live cell density. Using selective nucleic acid intercalating dyes like ethidium monoazide (EMA) and propidium monoazide (PMA) is one of the alternative approaches to detecting and quantifying viable cells by quantitative PCR. These compounds have the ability to penetrate only into dead cells with compromised membrane integrity and intercalate with DNA via their photoinducible azide groups and in turn inhibit DNA amplification during PCRs. PMA has been successfully used in different studies and microorganisms, but it has not been evaluated sufficiently for complex environmental samples such as biosolids. In this study, experiments were performed with Escherichia coli ATCC 25922 as the model organism and the uidA gene as the target sequence using real-time PCR via the absolute quantification method. Experiments with the known quantities of live and dead cell mixtures showed that PMA treatment inhibits PCR amplification from dead cells with over 99% efficiency. The results also indicated that PMA-modified quantitative PCR could be successfully applied to biosolids when the total suspended solids (TSS) concentration is at or below 2,000 mg·liter(-1).

Choice of indicator organism and library size considerations for phenotypic microbial source tracking by FAME profiling.

The primary objective of this study was to investigate the effects of choice of the indicator organisms on the accuracy of classifying the fatty acid methyl ester (FAME) profiles of the known-source library isolates. First, a known-source library containing the FAME profiles of Enterococcus isolates cultured from six different possible sources of microbial pollution was developed. A total of 511 Enterococcus isolates were profiled: 120 isolates from sewage samples representing humans; 69 from dairy and cattle cow; 74 from chicken; 76 from swine; 94 from deer; and 78 from waterfowl. Classification of known-source Enterococcus isolates into their respective host categories resulted with a 66% average rate of correct classification (ARCC) in a six-way discriminant analysis (DA). The ARCC increased to 75% when the individual hosts were pooled into larger categories of human, livestock, and wildlife. The accuracy was 80% when isolates of human origin were discriminated against those of non-human origins. Recently, several studies reported the ARCCs for various classification schemes associated with total coliform (TC), fecal coliform (FC), and Escherichia coli of the known-source isolates. When the accuracy of classification of Enterococcus isolates was compared to those reported for TC, FC, and E. coli isolates, the lowest ARCCs were associated with classification of E. coli isolates, the only species level indicator organism among the four compared. It was found that the degree of discrimination increases as the indicator becomes more inclusive of bacteria from different genus. In addition, random cluster formation analysis indicates that known-source libraries with isolate numbers between 300 and 500 might be sufficient for MST by FAME.

Changes in antibiotic resistance patterns of Escherichia coli during domestic wastewater treatment.

Changes in antibiotic resistance of Escherichia coli in the different stages of conventional domestic wastewater treatment were investigated. Over two years, more than 3500 E. coli isolates from four stages of the wastewater treatment process were tested for resistance to six different antibiotics. The percent resistance of bacteria from any of the stages was highly variable in different samples. Because of this variability, no statistically significant difference was found in the overall percent resistance of E. coli from influent to effluent. When comparing different stages within samples, however, there seemed to be an increase in resistance to ampicillin and amoxicillin between the raw influent and primary effluent. In addition, the percent of isolates with multiple antibiotic resistance, resistant to more than one and less than five antibiotics, and highly multiple antibiotic resistance, resistant to at least five antibiotics, increased through the treatment process.

Proposal of Viridibacillus gen. nov. and reclassification of Bacillus arvi, Bacillus arenosi and Bacillus neidei as Viridibacillus arvi gen. nov., comb. nov., Viridibacillus arenosi comb. nov. and Viridibacillus neidei comb. nov.

A polyphasic study was undertaken to clarify the taxonomic position of endospore-forming strains 433-D9, 433-E17 and 121-X1. BOX-PCR-generated fingerprints indicated that they may be members of a single species. 16S rRNA gene sequence similarity demonstrated that a representative of this group, 433-D9, is affiliated closely with Bacillus arvi DSM 16317(T) (100 %), Bacillus arenosi DSM 16319(T) (99.8 %) and Bacillus neidei NRRL BD-87(T) (97.1 %). Sequence similarities revealed Bacillus pycnus NRRL NRS-1691(T) and several Kurthia species as the next nearest relatives. DNA-DNA hybridization results showed that strain 433-D9 is a member of B. arvi. Detection of l-Lys-d-Asp-based peptidoglycan in strain 433-D9, B. arvi DSM 16317(T) and B. arenosi DSM 16319(T) was in agreement with their close relationship, but differentiated these strains from B. neidei NRRL BD-87(T) and B. pycnus NRRL NRS-1691(T), for which l-Lys-d-Glu was reported. A similar quinone system was detected in strains 433-D9, 433-E17, 121-X1, B. arvi DSM 16317(T), B. arenosi DSM 16319(T) and B. neidei NRRL BD-87(T). This system, unusual for bacilli, consisted of the major compound menaquinone MK-8 (69-80 %) and moderate amounts of MK-7 (19-30 %). This observation was in contrast to the predominance of MK-7 of the closest relative B. pycnus NRRL NRS-1691(T), as also reported for representatives of the closely related non-endospore-forming genus Kurthia. Strains 433-D9, B. arvi DSM 16317(T) and B. arenosi DSM 16319(T) exhibited homogeneous and discriminative polar lipid profiles and fatty acid profiles consisting of major acids i-C(15 : 0) and ai-C(15 : 0) and moderate amounts of i-C(17 : 1)omega10c and i-C(17 : 1) I/ai-C(17 : 1) B that discriminated them from closely related strains such as B. neidei NRRL BD-87(T). On the basis of clear-cut discriminative chemotaxonomic markers, we propose strains 433-D9, 433-E17 and 121-X1, B. arvi DSM 16317(T), B. arenosi DSM 16319(T) and B. neidei NRRL BD-87(T) to be reclassified within a separate genus. For this new taxon, we propose the name Viridibacillus gen. nov., and we propose the reclassification of Bacillus arvi, Bacillus arenosi and Bacillus neidei as Viridibacillus arvi gen. nov., comb. nov. (the type species of Viridibacillus, with the type strain DSM 16317(T) =LMG 22165(T)), Viridibacillus arenosi comb. nov. (type strain DSM 16319(T) =LMG 22166(T)) and Viridibacillus neidei comb. nov. (type strain NRRL BD-87(T) =DSM 15031(T) =JCM 11077(T)).

Phenotypic characterization of Escherichia coli through whole-cell fatty acid profiling to investigate host specificity.

The objective of the study was to investigate whole-cell fatty acid methyl ester (FAME) profiles of 605 Escherichia coli isolates to determine their host specificity. The isolates were cultured from six possible sources of fecal pollution; 180 isolates from sewage, 85 from dairy cow, 98 from chicken, 76 from swine, 94 from deer, and 72 from waterfowl, mostly geese and ducks. The FAME profiles were presented as the relative masses of 12 FAMEs identified in the isolates and it was found that none of the six hosts carried a "signature" FAME, a FAME that is uniquely associated with a particular host category. However, two-sample t-test analyses indicated that the mean relative masses of seven FAMEs out of the 12 identified showed statistically significant differences (95% confidence interval) between isolates of human and non-human origins. In addition, a linear discriminant function based on mean relative mass variations in individual FAMEs classified the known-source isolates into their respective host categories with a 47.6% average rate of correct classification (ARCC) in a six-way discriminant analysis. The ARCC increased to 61.3% when the individual hosts were pooled into larger categories of human, livestock, and wildlife. The accuracy was 75.5% when isolates of human origin were discriminated against those of non-human origins. Random cluster formation analysis indicated that the library size was sufficient to prevent random grouping among the isolates.

Thermophilic aerobic granular biomass for enhanced settleability.

Aerobic biological wastewater treatment at thermophilic (ca. 55 degrees C) temperatures notoriously produces biomass that flocculates poorly or not at all. Contrary to this, thermophilic aerobic biomass that settled well in sequencing batch reactors was cultured with sludge volume index (SVI) values as low as 60mL/g. A mixture of granular and flocculant biomass resulted when closed reactors were sparged with recirculated reactor headspace gas containing some air, whereas a conventionally aerated control reactor sparged with air alone contained dispersed growth that did not flocculate. Maximum granule diameter was from 1.2 to 1.9mm, and granule resistance to disintegration was comparable to aerobic mesophilic granules. Two bacteria were isolated and identified as Anoxybacillus flavothermus and Pseudoxanthomonas taiwanensis as determined by partial 16S rDNA sequencing. Anoxybacilli species are alkaliphilic or alkalitolerant, with the type species having an obligate requirement for carbonate, even when grown on glucose. We postulate that high alkalinity and CO(2) may select for a population of aerobic thermophilies that flocculates and granulates.

Bioaugmenting anaerobic digestion of biosolids with selected strains of Bacillus, Pseudomonas, and Actinomycetes species for increased methanogenesis and odor control.

The objective of this study was to evaluate the effects of bioaugmenting anaerobic biosolids digestion with a commercial product containing selected strains of bacteria from genera Bacillus, Pseudomonas, and Actinomycetes, along with ancillary organic compounds containing various micronutrients. Specifically, the effects of the bioaugment in terms of volatile solids destruction and generation and fate of odor-causing compounds during anaerobic digestion and during storage of the digested biosolids were studied. Two bench-scale anaerobic digesters receiving primary and secondary clarifier biosolids from various full-scale biological wastewater treatment plants were operated. One of the digesters received the bioaugment developed by Organica Biotech, while the other was operated as control. The bioaugmented digester generated 29% more net CH(4) during the 8 weeks of operation. In addition, the average residual propionic acid concentration in the bioaugmented digester was 54% of that in the control. The monitoring of two organic sulfide compounds, methyl mercaptan (CH(3)SH) and dimethyl sulfide (CH(3)SCH(3)), clearly demonstrated the beneficial effects of the bioaugmentation in terms of odor control. The biosolids digested in the bioaugmented digester generated a negligible amount of CH(3)SH during 10 days of post-digestion storage, while CH(3)SH concentration in the control reached nearly 300 ppm(v) during the same period. Similarly, peak CH(3)SCH(3) generated by stored biosolids from the bioaugmented digester was only 37% of that from the control.

Toxic effects of thiol-reactive compounds on anaerobic biomass.

The objective of this study was to characterize the toxic effects of three well known thiol-reactive electrophilic compounds, N-ethylmaleimide (NEM), pentachlorophenol (PCP) and 1-chloro-2,4-dinitrobenzene (CDNB) on anaerobic biotransformation process. The work was part of a larger investigation on potassium efflux as a possible response mechanism of anaerobic microorganisms to the presence of thiol-reactive organic compounds and the interference of such compounds on the reductive dehalogenation process. Using anaerobic toxicity assay (ATA) and granular anaerobic biomass from a full-scale upflow anaerobic sludge blanket (UASB) reactor, inhibitory concentrations of these compounds that reduced the microbial activity of granular biomass to 50% of a control (IC50) were determined to be 592, 0.97, and 450 mg/l for NEM, PCP, and CDNB, respectively. Toxicity of NEM was also tested on anaerobic biomass from a municipal wastewater treatment plant digester and slightly lower IC50 of 532 mg/l was obtained. The results presented here indicate that anaerobic biomass can acclimate to the three thiol-reactive compounds studied and recover from inhibition as long as the toxicant concentration is below a threshold level. That threshold concentration was found to be 500 mg/l for NEM on biomass from the municipal digester, 1 mg/l for PCP, and 500 mg/l for CDNB, both on granular biomass. Granular anaerobic biomass showed recovery even at NEM concentrations of 1000 mg/l.

Microbial source tracking using host specific FAME profiles of fecal coliforms.

The objective of this study was to investigate the host-specific differences in fatty acid methyl ester (FAME) profiles of fecal coliforms (FC). A known-source library was constructed with 314 FC isolates cultured from 6 possible sources of fecal pollution; 99 isolates from sewage; 29 from bovine; 29 from poultry; 50 from swine; 46 from waterfowl; and 61 from deer. It was found that the hydroxy FAMEs 12:0 2 OH, 12:03 OH, and 14:02 OH were exclusively associated with isolates of human origin. On the other hand, 3 saturated FAMEs, 10:0, 15:0, and 18:0 were found only in isolates from non-human sources, 15:0 being associated with livestock samples only. In addition to the presence of these signature FAMEs, the mean relative masses of 16:1 omega7c and 16:1 ISO/14:03 OH were significantly different between the isolates of human and non-human origins. A linear discriminant function differentiated FC isolates of human origin from those of livestock and wildlife origin at 99% accuracy. These results strongly suggest that the FAME profiles of FC show statistically significant host specificity and may have the potential to be used as a phenotypic microbial source tracking tool.