Anaerobic Degradation Of Phenol Research Articles

Microbiological insights into anaerobic phenol degradation mechanisms and bulking phenomenon in a mesophilic upflow anaerobic sludge blanket reactor in long-term operation

In this study, a long-term operation of 2,747 days was conducted to evaluate the performance of the upflow anaerobic sludge blanket (UASB) reactor and investigated the degradation mechanisms of high-organic loading phenol wastewater. During the reactor operation, the maximum chemical oxygen demand (COD) removal rate of 6.1 ± 0.6 kg/m3/day under 1,680 mg/L phenol concentration was achieved in the mesophilic UASB reactor. After a significant change in the operating temperature from 24.0 ± 4.1 °C to 35.9 ± 0.6 °C, frequent observations of floating and washout of the bloated granular sludge (novel types of the bulking phenomenon) were made in the UASB reactor, suggesting that the change in operating temperature could be a trigger for the bulking phenomenon. Through the metagenomic analysis, phenol degradation mechanisms were predicted that phenol was converted to 4-hydroxybenzoate via two possible routes by Syntrophorhabdaceae and Pelotomaculaceae bacteria. Furthermore, the degradation of 4-hydroxybenzoate to benzoyl-CoA was carried out by members of Syntrophorhabdaceae and Smithellaceae. In the bulking sludge, a predominant presence of Nanobdellota, belonging to DPANN archaea, was detected. The metagenome-assembled genome of the Nanobdellota lacks many biosynthetic pathways and has several genes for the symbiotic lifestyle such as trimeric autotransporter adhesin-related protein. Furthermore, the Nanobdellota have significant correlations with several methanogenic archaea that are predominantly present in the UASB reactor. Considering the results of this study, the predominant Nanobdellota may negatively affect the growth of the methanogens through the parasitic lifestyle and change the balance of microbial interactions in the granular sludge ecosystem.

Microbial electricity-driven anaerobic phenol degradation in bioelectrochemical systems

Microbial electrochemical technologies have been extensively employed for phenol removal. Yet, previous research has yielded inconsistent results, leaving uncertainties regarding the feasibility of phenol degradation under strictly anaerobic conditions using anodes as sole terminal electron acceptors. In this study, we employed high-performance liquid chromatography and gas chromatography-mass spectrometry to investigate the anaerobic phenol degradation pathway. Our findings provide robust evidence for the purely anaerobic degradation of phenol, as we identified benzoic acid, 4-hydroxybenzoic acid, glutaric acid, and other metabolites of this pathway. Notably, no typical intermediates of the aerobic phenol degradation pathway were detected. One-chamber reactors (+0.4 V vs. SHE) exhibited a phenol removal rate of 3.5 ± 0.2 mg L−1 d−1, while two-chamber reactors showed 3.6 ± 0.1 and 2.6 ± 0.9 mg L−1 d−1 at anode potentials of +0.4 and + 0.2 V, respectively. Our results also suggest that the reactor configuration certainly influenced the microbial community, presumably leading to different ratios of phenol consumers and microorganisms feeding on degradation products.

Metagenomic Binning Revealed Microbial Shifts in Anaerobic Degradation of Phenol with Hydrochar and Pyrochar

Phenolic compounds, which are difficultly degraded, are one of the main toxic threats faced in the anaerobic digestion (AD) process. It has previously been reported that hydrochar/pyrochar produced by the hydrothermal liquefaction/pyrolysis of biomass can enhance AD by promoting direct interspecific electron transfer (DIET). The present study investigated the effects of different hydrochars and pyrochars on the anaerobic degradation of phenol and provided deep insights into the related micro-organisms at the species level through genome-centric metagenomic analysis. Compared with the control experiment, the addition of hydrochar and pyrochar shortened the lag time. However, hydrochar created a large increase in the maximum methane production rate (Rm) (79.1%) compared to the control experiments, while the addition of pyrochar decreased Rm. Metagenomic analysis showed that the addition of carbon materials affected the relative abundance of genes in the phenol anaerobic degradation pathway, as well as the species and relative abundance of phenol degrading micro-organisms. The relative abundance of key genes for phenol degradation, such as bsdB, bamB, oah, etc., under the action of hydrochar was higher than those under the action of pyrochar. In addition, hydrochar-enriched phenol degradation-related bacteria (Syntrophus aciditrophicus, etc.) and methanogen (Methanothrix soehngenii, etc.). These micro-organisms might improve the phenol degradation efficiency by promoting DIET. Therefore, hydrochar had a more significant effect in promoting anaerobic degradation of phenol.

Mapping substrate use across a permafrost thaw gradient

Permafrost thaw in northern peatlands is likely to create a positive feedback to climate change, as microbes transform soil carbon (C) into carbon dioxide (CO2) or methane (CH4). While the microbiome's encoded C-processing potential changes with thaw, the impact on substrate utilization and gas emissions is less well characterized. We therefore examined microbial C-cycling dynamics from a partially thawed Sphagnum-dominated bog to a fully thawed sedge-dominated fen in Stordalen Mire (68.35°N, 19.05°E), Sweden. We profiled C substrate utilization diversity and extent by Biolog Ecoplates™, then tested substrate-specific hypotheses by targeted additions (of glucose, the short chain fatty acids (SCFAs) acetate and butyrate, and the organic acids galacturonic acid and p-hydroxybenzoic acid, all at field-relevant concentrations) under anaerobic conditions at 15 °C. In parallel we characterized microbiomes (via 16S rRNA amplicon sequencing and quantitative polymerase chain reaction) and C gas emissions. The fen exhibited a higher substrate use diversity and faster rate of overall substrate utilization than in the bog, based on Biolog Ecoplate™ incubations. Simple glucose additions (akin to a positive control) to peat microcosms fueled fermentation as expected (reflected in enriched fermenter lineages, their inferred metabolisms, and CO2 production), but also showed potential priming of anaerobic phenol degradation in the bog. Addition of SCFAs to bog and fen produced the least change in lineages and in CO2, and modest suppression of CH4 primarily in the fen, attributed to inhibition. Addition of both organic acids greatly increased the CO2:CH4 ratio in the deep peats but had distinct individual gas dynamics and impacts on microbiota. Both organic acids appeared to act as both C source and as a microbial inhibitor, with galacturonic acid also likely playing a role in electron transfer or acceptance. Collectively, these results support the importance of aboveground-belowground linkages - and in particular the role of Sphagnum spp.- in supplying substrates and inhibitors that drive microbiome assembly and C processing in these dynamically changing systems. In addition, they highlight an important temporal dynamic: responses on the short time scale of incubations (which would reflect transition conditions in the field) differ from those evident at the longer scales of habitat transition, in ways consequential to C gas emissions. In the short term, substrate addition response reflected microbiome legacy (e.g., bog communities were slower to process C and better tolerated inhibitors than fen communities) but led to little overall increase in C gas production (and a high skew to CO2). At the longer time scale of bog and fen thaw stages (which are used to represent these systems in models) the concomitant shifts in plants, hydrology and microbiota attenuate microbiome legacy impacts on substrate processing and C gas emissions over time. As habitat transition areas expand under accelerating change, we hypothesize an increased role of microbiome legacy in the landscape overall, leading to a lag in the increase of CH4 emissions expected from fen expansion.

Subsurface hydrocarbon degradation strategies in low- and high-sulfate coal seam communities identified with activity-based metagenomics

Environmentally relevant metagenomes and BONCAT-FACS derived translationally active metagenomes from Powder River Basin coal seams were investigated to elucidate potential genes and functional groups involved in hydrocarbon degradation to methane in coal seams with high- and low-sulfate levels. An advanced subsurface environmental sampler allowed the establishment of coal-associated microbial communities under in situ conditions for metagenomic analyses from environmental and translationally active populations. Metagenomic sequencing demonstrated that biosurfactants, aerobic dioxygenases, and anaerobic phenol degradation pathways were present in active populations across the sampled coal seams. In particular, results suggested the importance of anaerobic degradation pathways under high-sulfate conditions with an emphasis on fumarate addition. Under low-sulfate conditions, a mixture of both aerobic and anaerobic pathways was observed but with a predominance of aerobic dioxygenases. The putative low-molecular-weight biosurfactant, lichysein, appeared to play a more important role compared to rhamnolipids. The methods used in this study—subsurface environmental samplers in combination with metagenomic sequencing of both total and translationally active metagenomes—offer a deeper and environmentally relevant perspective on community genetic potential from coal seams poised at different redox conditions broadening the understanding of degradation strategies for subsurface carbon.

Analysis of Carbon and Hydrogen Stable Isotope Ratios of Phenolic Compounds: Method Development and Biodegradation Applications

Phenolic compounds are priority water pollutants of both natural and anthropogenic origins. Distinguishing the sources and degradation pathways of phenolic compounds in the environment is essential for sustainable water management. In this study, we have developed and validated methods based on gas chromatography–isotope ratio mass spectrometry for analysis of hydrogen and carbon isotope ratios of phenol and cresols. For carbon stable isotope analysis, solid-phase extraction was applied, whereas liquid–liquid extraction, followed by derivatization with trifluoroacetic anhydride, was used for hydrogen stable isotope analysis. The methods were verified in biodegradation experiments using p-cresol and phenol as substrates. Methyl group oxidation of p-cresol by Pseudomonas pseudoalcaligenes and Aromatoleum aromaticum resulted in the strong hydrogen (−76 ± 2 and −103 ± 2 mUr, respectively) and varying carbon isotopic fractionation (−2.1 ± 0.1 and −1.0 ± 0.1 mUr, respectively), leading to distinguishable two-dimensional plots for hydrogen versus carbon isotope ratios, indicating slightly different enzymatic reactions. For anaerobic phenol degradation by Desulfosarcina cetonica, the absence of hydrogen isotopic fractionation indicates adenosine triphosphate-dependent carboxylation as the initial step of the biodegradation pathway. The methods developed in this study are useful to detect biological degradation of phenolic compounds in highly contaminated aqueous environmental systems.

Anaerobic biodegradation of phenol in wastewater treatment: achievements and limits

Anaerobic biodegradation of toxic compounds found in industrial wastewater is an attractive solution allowing the recovery of energy and resources but it is still challenging due to the low kinetics making the anaerobic process not competitive against the aerobic one. In this review, we summarise the present state of knowledge on the anaerobic biodegradation process for phenol, a typical target compound employed in toxicity studies on industrial wastewater treatment. The objective of this article is to provide an overview on the microbiological and technological aspects of anaerobic phenol degradation and on the research needs to fill the gaps still hindering the diffusion of the anaerobic process. The first part is focused on the microbiology and extensively presents and characterises phenol-degrading bacteria and biodegradation pathways. In the second part, dedicated to process feasibility, anaerobic and aerobic biodegradation kinetics are analysed and compared, and strategies to enhance process performance, i.e. advanced technologies, bioaugmentation, and biostimulation, are critically analysed and discussed. The final section provides a summary of the research needs. Literature data analysis shows the feasibility of anaerobic phenol biodegradation at laboratory and pilot scale, but there is still a consistent gap between achieved aerobic and anaerobic performance. This is why current research demand is mainly related to the development and optimisation of powerful technologies and effective operation strategies able to enhance the competitiveness of the anaerobic process. Research efforts are strongly justified because the anaerobic process is a step forward to a more sustainable approach in wastewater treatment.Key points• Review of phenol-degraders bacteria and biodegradation pathways.• Anaerobic phenol biodegradation kinetics for metabolic and co-metabolic processes.• Microbial and technological strategies to enhance process performance.

Biochar and GAC intensify anaerobic phenol degradation via distinctive adsorption and conductive properties

The roles of biochar and granular activated carbon (GAC) in the enhancement of anaerobic phenol degradation were characterized through batch tests conducted at different phenol concentrations, coupled with adsorption kinetics, microbial community, and in-situ electrochemical analysis. Both biochar and GAC (15 g/L) led to markedly shorter lag times (t0) by adsorbing dissolved phenol, and faster maximum CH4 production rate (Rmax) by triggering direct interspecies electron transfer (DIET) during a two-stage (adsorption then degradation) anaerobic phenol degradation. The high adsorption capacity of GAC helped achieve a shorter t0, but less affected Rmax of subsequent phenol degradation. Compared with GAC, which showed higher conductivity but no redox activity, biochar exhibited higher electron exchange capacity (6.57 μmol e−/g). This higher electron exchange capacity stemmed from the diverse redox-active moieties, which resulted in a more efficient DIET. Meanwhile, the formation of wire-like appendages which linked the enriched DIET partners (such as Syntrophorhabdus and Methanosaeta) on biochar probably futher enhanced the electron transfer. However, hydrogenotrophic methanogenesis was still the main pathway for syntrophic phenol degradation in the suspended sludge. The in-situ analysis also confirmed that biochar and GAC acted as geobatteries and geoconductors, respectively, and that the stimulation of DIET was persistent.

Influence of particle size distribution on anaerobic degradation of phenol and analysis of methanogenic microbial community.

Sludge morphology considerably affects the mechanism underlying microbial anaerobic degradation of phenol. Here, we assessed the phenol degradation rate, specific methanogenic activity, electron transport activity, coenzyme F420 concentration, and microbial community structure of five phenol-degrading sludge of varying particle sizes (i.e., < 20, 20-50, 50-100, 100-200, and > 200μm). The results indicated an increase in phenol degradation rate and microbial community structure that distinctly correlated with an increase in sludge particle size. Although the sludge with the smallest particle size (< 20μm) showed the lowest phenol degradation rate (9.3mg COD·gVSS-1 day-1), its methanogenic activity with propionic acid, butyric acid, and H2/CO2 as substrates was the best, and the concentration of coenzyme F420 was the highest. The small particle size sludge did not contain abundant syntrophic bacteria or hydrogenotrophic methanogens, but contained abundant acetoclastic methanogens. Moreover, the floc sizes of the different sludge varied in important phenol-degrading bacteria and archaea, which may dominate the synergistic mechanism. This study provides a new perspective on the role of sludge floc size on the anaerobic digestion of phenol.

Denitrifying Microbial Community Structure and bamA Gene Diversity of Phenol Degraders in Soil Contaminated from the Coking Process.

Phenolic compounds are the dominant pollutants in soils contaminated by the coking industry. Ring opening by the hydroxylase gene (bamA) is the key step in the benzoyl-CoA degradation pathway under anaerobic conditions, and a broad spectrum of microorganisms possesses this functional gene, including denitrifiers. The present study analyzed the community structure of denitrifying bacteria and the diversity of the bamA gene for mixed cultures enriched from soil collected at a coking industrial site and then grown under nitrate-reducing conditions on phenol or p-hydroxybenzoate (4HBA), a key intermediate product of anaerobic phenol degradation. Illumina sequencing of the 16S rRNA gene showed different bacterial compositions between the two cultures. The dominant phyla were Proteobacteria, Armatimonadetes, and Planctomycetes in the phenol culture and Proteobacteria and Bacteroidetes in the 4HBA culture. Phylogenetic analysis further demonstrated that bamA genes were associated with four clusters of bacteria, three of known bacteria and one of uncultured bacteria. The diversity of the bamA gene differed from that reported in anaerobic aromatic degradation cultures, suggesting that these enriched cultures may contain new strains unique to coking-contaminated soils. The present study further validates the potential application of this functional gene as a marker for anaerobic biodegradation processes in enrichment cultures from contaminated soil.

Enhancing anaerobic degradation of phenol to methane via solubilizing Fe(III) oxides for dissimilatory iron reduction with organic chelates

The improved performances during anaerobic degradation of phenol to methane with Fe(OH)3 were usually inapparent, due to its lower solubility (unaccessible to dissimilatory iron reduction) and more positive reduction potential of Fe(III)/Fe(II) (unfavorable for enriching Fe(III)-reducing bacteria [IRBs]). In this study, citrate, the organic chelates, were used to solubilize Fe(III) with the aim of improving the phenol degradation and declining the reduction potential of Fe(III)/Fe(II). Results showed that, in the co-occurrence of citrate and Fe(OH)3, the degradation rates of phenol were about 1.3-fold rapider than that with sole Fe(OH)3. Analysis of cyclic voltammetry demonstrated that the reduction potential of Fe(III)/Fe(II) in the form of Fe(OH)3 (−0.41 to −0.28 V vs Ag/AgCl) declined to −0.61 to −0.41 V. As a result, the Fe(III)-reducing genera, such as Petrimonas and Shewanella, which held a great potential of proceeding syntrophic metabolism via direct interspecies electron transfer (DIET), were significantly enriched.

Key microbial populations involved in anaerobic degradation of phenol and p-cresol using different inocula

Key microbial populations involved in anaerobic degradation of phenol and p-cresol using different inocula

Enzymes involved in the anaerobic degradation of phenol by the sulfate-reducing bacterium Desulfatiglans anilini

BackgroundThe sulfate-reducing bacterium Desulfatiglans anilini can grow with phenol as sole source of carbon and energy under strictly anaerobic, sulfate-reducing conditions. In the nitrate-reducing bacterium Thauera aromatica, the enzymes involved in phenol degradation have been well elucidated, whereas the anaerobic phenol degradation pathway by D. anilini was not studied in detail yet.ResultsThe pathway of anaerobic phenol degradation by the sulfate-reducing bacterium Desulfatiglans anilini was studied by identification of genes coding for phenylphosphate synthase (encoded by pps genes) and phenylphosphate carboxylase (encoded by ppc genes) in the genome of D. anilini, by analysis of the transcription and translation of pps-ppc genes, and by measurement of phenylphosphate synthase activity in cell-free extracts of phenol-grown cells. The majority of genes involved in phenol degradation were found to be organized in one gene cluster. The gene cluster contained genes ppsα (phenylphosphate synthase alpha subunit), ppsβ (phenylphosphate synthase beta subunit), ppcβ (phenylphosphate carboxylase beta subunit), as well as 4-hydroxybenzoyl-CoA ligase and 4-hydroxylbenzoyl-CoA reductase-encoding genes. The genes ppsγ (phenylphosphate synthase gamma subunit), ppcα (phenylphosphate carboxylase alpha subunit) and ppcδ (phenylphosphate carboxylase delta subunit) were located elsewhere in the genome of D. anilini, and no obvious homologue of ppcγ (phenylphosphate carboxylase gamma subunit) was found in the genome. Induction of genes pps and ppc during growth on phenol was confirmed by reverse transcription polymerase chain reaction. Total proteome analysis revealed that the abundance of enzymes encoded by the gene cluster under study was much higher in phenol-grown cells than that in benzoate-grown cells. In in-vitro enzyme assays with cell-free extracts of phenol-grown cells, phenylphosphate was formed from phenol in the presence of ATP, Mg2+, Mn2+, K+ as co-factors.ConclusionsThe genes coding for enzymes involved in the anaerobic phenol degradation pathway were identified in the sulfate-reducing bacterium D. anilini. The results indicate that the first steps of anaerobic phenol degradation in D. anilini are phosphorylation of phenol to phenylphosphate by phenylphosphate synthase and carboxylation of phenylphosphate by phenylphosphate carboxylase.

Key microbial populations involved in anaerobic degradation of phenol and p-cresol using different inocula

Background: Anaerobic digestion is an alternative bioprocess used to treat effluents containing toxic compounds such as phenol and p-cresol. Selection of an adequate sludge as inoculum containing an adapted microbial consortium is a relevant factor to improve the removal of these pollutants. The objective of this study is to identify the key microorganisms involved in the anaerobic digestion of phenol and p-cresol and elucidate the relevance of the bamA gene abundance (a marker gene for aromatic degraders) in the process, in order to establish new strategies for inocula selection and improve the system's performance. Results: Successive batch anaerobic digestion of phenol and p-cresol was performed using granular or suspended sludge. Granular sludge in comparison to suspended sludge showed higher degradation rates both for phenol (11.3 ± 0.7 vs 8.1 ± 1.1 mg l-1 d-1) and p-cresol (7.8 ± 0.4 vs 3.7 ± 1.0 mg l-1 d-1). After three and four re-feedings of phenol and p-cresol, respectively, the microbial structure from both sludges was clearly different from the original sludges. Anaerobic digestion of phenol and p-cresol generated an abundance increase in Syntrophorhabdus genus and bamA gene, together with hydrogenotrophic and aceticlastic archaea. Analysis of results indicates that differences in methanogenic pathways and levels of Syntrophorhabdus and bamA gene in the inocula, could be the causes of dissimilar degradation rates between each sludge. Conclusions: Syntrophorhabdus and bamA gene play relevant roles in anaerobic degradation of phenolics. Estimation of these components could serve as a fast screening tool to find the most acclimatized sludge to efficiently degrade mono-aromatic compounds.

Enhanced anaerobic phenol degradation by conductive materials via EPS and microbial community alteration

Anaerobic biodegradation is a promising and economical process to remove phenol in the wastewater, although the reaction kinetics are often limited by the toxicity of phenol. This study proposed a strategy to accelerate phenol degradation and mitigate its inhibition on the bioprocess by adding conductive materials (CMs) into the anaerobic degradation system. The results showed that CMs could stimulate phenol degradation up to one–fold higher than that in control group. Certain extracellular polymeric substances (EPS) compounds that can act as electron shuttles, i.e. protein and humic substances, were greatly enriched with the existence of CMs. In particular, compared with control group, CMs supplemented groups had 2.3 and 10 to 20 folds low molecular weight protein in bound and soluble EPS, respectively. Carbon nanotube supplemented group exhibited 3 times higher humic substance in soluble EPS. The overall electron transport system activity in CMs groups was much higher than that in control groups. The addition of CMs enriched phenol degradation bacteria–Syntrophorhabdus, Brooklawnia, Treponema and Syntrophus, as well as electroactive methanogens–Methanosaeta. It is proposed that Syntrophus and Methanosaeta were the functional genera in methanogenic phenol degradation via extracellular electron transfer. This study revealed that the presence of CMs altered the EPS composition and microbial community of the system and made it become more favorable for extracellular electron transfer.

Specific methanogenic activity test for anaerobic treatment of phenolic wastewater

AbstractAnaerobic processes have been successfully used in the treatment of municipal and industrial wastewater. However, the effect of nutrients like nitrogen and phosphorus on specific methanogenic activity (SMA) in anaerobic treatment of wastewaters containing phenol has not been systematically investigated. Phenol is present in industrial effluents like coal gasification, coke production, pharmaceutical, pesticide, fertilizer, dye manufacturing, synthetic chemical, and pulp and paper. This paper presents experimental study on determination of SMA using phenol as a sole substrate and the present work is being compared with the studies carried out by various researchers using different substrates. SMA test was carried out immediately after seeding the upflow anaerobic sludge blanket reactor with sludge in order to determine the initial phenol biodegradation during the start-up phase of the reactor. Anaerobic degradation of phenol to methane takes place via benzoate and then to acetate. Therefore, the SM...

Phylogenetic and functional diversity of metagenomic libraries of phenol degrading sludge from petroleum refinery wastewater treatment system

In petrochemical refinery wastewater treatment plants (WWTP), different concentrations of pollutant compounds are received daily in the influent stream, including significant amounts of phenolic compounds, creating propitious conditions for the development of particular microorganisms that can rapidly adapt to such environment. In the present work, the microbial sludge from a refinery WWTP was enriched for phenol, cloned into fosmid vectors and pyrosequenced. The fosmid libraries yielded 13,200 clones and a comprehensive bioinformatic analysis of the sequence data set revealed a complex and diverse bacterial community in the phenol degrading sludge. The phylogenetic analyses using MEGAN in combination with RDP classifier showed a massive predominance of Proteobacteria, represented mostly by the genera Diaphorobacter, Pseudomonas, Thauera and Comamonas. The functional classification of phenol degrading sludge sequence data set generated by MG-RAST showed the wide metabolic diversity of the microbial sludge, with a high percentage of genes involved in the aerobic and anaerobic degradation of phenol and derivatives. In addition, genes related to the metabolism of many other organic and xenobiotic compounds, such as toluene, biphenyl, naphthalene and benzoate, were found. Results gathered herein demonstrated that the phenol degrading sludge has complex phylogenetic and functional diversities, showing the potential of such community to degrade several pollutant compounds. This microbiota is likely to represent a rich resource of versatile and unknown enzymes which may be exploited for biotechnological processes such as bioremediation.

Degradation of phenolic compounds in aerobic and anaerobic landfills: a pilot scale study

The aim of this study was to investigate the aerobic and anaerobic degradation of phenol and its derivatives in aerobic and anaerobic landfills. Phenolic compounds were extracted from leachate samples using the solid phase micro-extraction method. In this study, analysis of the 24 phenolic compounds included in the standard mixture and the change in the concentrations over time of 23 of the 24 compounds found in the calibration mix standard were determined in both aerobic and anaerobic landfill reactors. It can be concluded that faster and complete removal of phenol, chlorophenol, dichlorophenols, and trichlorophenol were achieved in the aerobic landfill while aerobic treatment was less effective on tetrachlorophenol and pentachlorophenol. In the anaerobic landfill, anaerobic reductive dechlorination probably occurred from all the highly chlorinated phenols and resulted in the accumulation of phenol and chlorophenol. The phenol could not be further degraded because the anaerobic methanogenic phase did not start during the 150 days of operation in an anaerobic landfill reactor. Nitrophenols can be degraded rapidly under aerobic conditions. These compounds are degraded to amino groups in the first step and then these amino groups are degraded to methane and CO(2) under anaerobic conditions. Although the degradation could not reach the methanogenic phase in anaerobic landfill reactor during the operational period, it is indicated that nitrophenol concentrations decreased in the anaerobic reactor. This is revealed as a result of the formation of the amino groups.

Development and validation of a simplified model for the anaerobic degradation of phenol

The anaerobic treatment of phenolic wastewater has demonstrated to be a suitable biological system, for that reason, a large number of systems have been implemented in a lab/pilot scale, several industrial plants have also been developed. Despite of this, there is a lack of modeling applications within these systems. In order to enhance the anaerobic treatment of this kind of water, a simplified model of 2 populations and 3 reactions was developed and implemented. The parameter calibration and the model validation were carried out with experimental data obtained from an Anaerobic Sequencing Batch Reactor treating phenolic wastewater through two different operational strategies: sequential batches with a co-substrate and sequential fed-batches without a co-substrate. The model predicted the reactors performance accurately for the different experimental conditions tested. Therefore, the theoretical basis of the model is, in general terms, valid, and its utilization to predict the reactors performance or in control purposes is feasible.

Degradation of Phenol via Phenylphosphate and Carboxylation to 4-Hydroxybenzoate by a Newly Isolated Strain of the Sulfate-Reducing BacteriumDesulfobacterium anilini

A sulfate-reducing phenol-degrading bacterium, strain AK1, was isolated from a 2-bromophenol-utilizing sulfidogenic estuarine sediment enrichment culture. On the basis of phylogenetic analysis of the 16S rRNA gene and DNA homology, strain AK1 is most closely related to Desulfobacterium anilini strain Ani1 (= DSM 4660(T)). In addition to phenol, this organism degrades a variety of other aromatic compounds, including benzoate, 2-hydroxybenzoate, 4-hydroxybenzoate, 4-hydroxyphenylacetate, 2-aminobenzoate, 2-fluorophenol, and 2-fluorobenzoate, but it does not degrade aniline, 3-hydroxybenzoate, 4-cyanophenol, 2,4-dihydroxybenzoate, monohalogenated phenols, or monohalogenated benzoates. Growth with sulfate as an electron acceptor occurred with acetate and pyruvate but not with citrate, propionate, butyrate, lactate, glucose, or succinate. Strain AK1 is able to use sulfate, sulfite, and thiosulfate as electron acceptors. A putative phenylphosphate synthase gene responsible for anaerobic phenol degradation was identified in strain AK1. In phenol-grown cultures inducible expression of the ppsA gene was verified by reverse transcriptase PCR, and 4-hydroxybenzoate was detected as an intermediate. These results suggest that the pathway for anaerobic degradation of phenol in D. anilini strain AK1 proceeds via phosphorylation of phenol to phenylphosphate, followed by carboxylation to 4-hydroxybenzoate. The details concerning such reaction pathways in sulfidogenic bacteria have not been characterized previously.