Microbial community structure in anaerobic degradation of terephthalate and phenol 3

Introduction Chapter Introduction Introduction 1.1 Background With the advancement of technology in the 20th century, synthetic products such as petrochemical-related materials have gained their popularity due to their durability in physical properties and aesthetics in appearance. As a result, the demand of raw materials such as terephthalate and phenol for the manufacturing of plastic or petrochemical-related products has increased, and the production of terephthalate- and phenol-containing wastewaters has also increased over the years. Purified terephthalate (PTA, or 1,4-benzenedicarboxylic acid) is an important raw material used in the manufacture of various plastic products. The most well-known application of PTA is the production of polyethylene terephthalate (PET) bottles for carbonated drinks (Park & Sheehan, 1996). Other applications include polyester films, textile fibers, adhesive, coatings and packing materials (Franck & Stadelhofer, 1988). PTA is mainly produced in Asia (South Korea, Taiwan, China, Indonesia, Japan, Thailand and Malaysia), and the amount accounts for approximately 66% of total PTA production worldwide. The amount produced was approximately 35 million tons in 2006, and is increased at an annual rate of to 10% (Razo-Flores et al., 2006). During the production of PTA, high-strength wastewater containing terephthalate as the main component and other organic compounds like acetate, benzoate and p-toluate (para-toluate) is produced and discharged. Typically, one ton of PTA produced is accompanied by about 2.5−4.5 m3 of wastewater at a concentration equivalent to 20−40 kg chemical oxygen demand (COD)/m3 (Bushway & Gilman, 1986; Kleerebezem et al., 1999c; Shelley, 1991; Vanduffel, 1993). Introduction Like PTA, phenol is used as the raw material for the production of various resins like phenolic, epoxy, polycarbonate and polyamide for plywood adhesive, construction, automotive and appliance industries (Kirk-Othmer, 1978). The amount produced was estimated to be million tons in 2001 and is increased at an annual rate of 6%. Phenol is also a common constituent in the wastewaters generated from phenol manufacturing processes, petrochemical-related processes, coal gasification, coke ovens and oil refining processes. The concentration of phenol and phenolic compounds in wastewaters is approximately 1.0−1.7 x 104 mg/L, and the COD contributed by phenol and phenolic compounds ranges from 40 to 80% of the total COD in the wastewaters (Veeresh et al., 2005). With the advantages of cost-saving (low energy and nutrients requirements), effective removal, and energy recovery (methane), anaerobic biological treatments processes have gradually replaced aerobic processes as an attractive alternative for treating toxic and poorly biodegradable wastewaters. Since the late 1980s, various types of full-scale anaerobic reactors including the downflow stationary fixed film (DSFF), hybrid, anaerobic contact (AC), upflow anaerobic sludge bed (UASB), downflow anaerobic filter (DAF), internal circulation (IC), expanded granular sludge bed (EGSB) and upflow anaerobic filter (UAF) systems have been demonstrated to effectively treat PTA wastewater (Macarie, 2000). During these anaerobic treatment, degradation of terephthalate has been suggested as the rate limiting step (Kleerebezem et al., 1997; Kleerebezem et al., 2005). Phenol is highly toxic and a strong growth inhibitor for microorganisms, but it has been successfully treated by a full-scale and a number of Introduction laboratory-scale anaerobic reactors (Lao, 2002; Suidan et al., 1983a; Veeresh et al., 2005). During the anaerobic methanogenic treatment, terephthalate and phenol are degraded through methanogenic metabolism in the absence of alternative electron acceptors (e.g., sulfates, nitrates and oxidized forms metals). This process requires the interaction between syntrophic fermentative bacteria and methanogenic archaea (methanogens) so that the degradation of terephthalate and phenol can be coupled with the hydrogen- and acetate-consuming reactions to overcome the thermodynamic barrier in the initial step (positive Gibbs free energy, ∆Go') (Fig. 1.1), and the intermediates can be converted to the final gaseous products (CH4 and CO2). Figure 1.1 conditions. Proposed terephthalate and phenol degradation under methanogenic 1.2 Problem statements Due to the recent awareness of environmental protection, more stringent environmental legislation has been implemented to control the discharge of potentially harmful compounds in many countries. Phenol is a carcinogenic compound, and can affect Introduction aquatic life at concentrations >1 mg/L (Autenrieth et al., 1991), whereas the toxicity of terephthalate remains unclear. Thus, the release of these compounds in environments has attracted great environmental and public health concerns in recent years, and there is a strong need to effectively treat these terephthalate- and phenol-containing wastewaters before discharge. Over the last 20 years, although empirical know-how of anaerobic treatment technologies utilimately reaches the good performance on the treatments of these wastewaters, these still face a number of challenges, as described below, for improvement. (A) Limited application of treatment processes for PTA wastewater at temperatures above 37°C Anaerobic processes for treating PTA wastewater have been successfully achieved under mesophilic (30−37ºC) conditions. As the production of PTA is carried out under high pressure and temperature, the PTA wastewaters are usually generated at high temperatures (40−60ºC). Therefore, additional time to store them in an equalization tank to cool down the wastewater before the mesophilic anaerobic treatment processes is necessary. It further suggests that anaerobic treatment processes under high temperatures (>37ºC) conditions can be an attractive alternative. So far, the development of thermophilic anaerobic treatment is relatively limited. Thermophilic treatments are reported to generally have much higher specific organic removal rates than the mesophilic treatments (van Lier et al., 1997), but it is relatively difficult to establish a stable microbial consortium in the thermophilic (55°C) terephthalate-degrading reactor (Kleerebezem et al., 1999c). This is attributed to a low number of thermophilic Introduction terephthalate-degrading microorganisms in the original seed sludge, and inappropriate operational conditions to enrich the microbial consortia (Kleerebezem et al., 1999c). Until now, the biodegradability of terephthalate and the responsible microorganisms at temperatures above 37°C is poorly known and further studies are necessary. (B) Limited information on phenol-degrading microbial consortia Anaerobic biological processes have been demonstrated to efficiently treat phenol-containing wastewaters at organic loadings (6−8 kg COD/m3 • day) in the laboratory-scale bioreactors (Veeresh et al., 2005). Prior to reaching the extraordinary performance (>90% removal of phenol), it requires a long start-up time (~300 days) to acclimatize the suitable microbial populations from seed sludge, to develop efficient phenol-degrading capability as well as particular microbial interaction and spatial orientation that can facilitate the rapid exchange of nutrients and products (Veeresh et al., 2005). Like the terephthalate-degrading methanogenic consortium, phenol degradation under thermophilic conditions is reported to be relatively difficult and unstable as compared to that under mesophilic conditions (Fang et al., 2006; Karlsson et al., 1999). As a result, the development of thermophilic techniques to treat phenol-containing wastewater is relatively limited. A better understanding of the phenol-degrading microbial community could provide crucial information for better selection of seeding sludge to shorten the start-up time in the mesophilic processes, and facilitate the development of thermophilic processes. So far, the phenol-degrading methanogenic microbial consortia was only characterized for biomass samples taken from a laboratory reactor operated at a temperature at 26°C (Fang et al., 2004; Zhang et al., 2005). The Introduction microbial populations responsible for phenol degradation under mesophilic and thermophilic conditions have not been fully identified and characterized. (C) Limited information on full-scale reactor to treat phenol-containing wastewater Many anaerobic laboratory-scale bioreactors have been successfully used to treat phenol-containing wastewaters. However, most of them were fed with synthetic wastewater and had better control of the operational parmeters (e.g., loading rate, recycle rate, temperature and hydraulic retention time [HRT]). These laboratory-scale bioreactors may not provide the same environmental conditions as the full-scale bioreators. As a result, the microbial populations found in the laboratory-scale bioreactors or enrichment cultures may not fully represent those in full-scale bioreactors treating wastewaters containing phenol. So far, only one full-scale UASB reactor has been constructed and operated to treat the wastewater from phenol production under mesophilic conditions since 1986 (Macarie, 2000). To facilitate the development of full-scale processes, a better understanding of the phenol-degrading microbial community in a full-scale plant is necessary. (D) Limitation of conventional culture-dependent approaches The bioreactor where the degradation of organic pollutants takes place traditonally referred to as a “black box”. In the past 10 years, microbial populations inside wastewater treatment plants have been extensively studied in order to correlate their identity, abundance and dynamics to the reactor performance. In anaerobic wastewater treatment systems for treating terephthalate- or phenol-containing wastewaters, microorganisms Introduction play as the catalysts to degrade the substances. However, owing to the limitations associated with the conventional techniques (time-consuming and low culturability of environmental microorganisms) and the difficulty of isolating syntrophic bacteria, only two mesophilic terephthalate-degrading bacteria (Pelotomaculum terephthalicum and Pelotomaculum isophthalicum) (Qiu et al., 2004; Qiu et al., 2006) and one phenol-transforming bacteria (Cryptanaerobacter phenolicus) (Juteau et al., 2005) have been isolated. The presence of these isolates P. isophthalicum, P. terephthalicum and C. phenolicus in the laboratory-, pilot- and full-scale terephthalate- or phenol-degrading reactors remains unknown. It is almost impossible to describe microbial community in the reactors based on the information of limited isolate strains available so far, and the isolates may not even represent as the main population involved in terephthalate or phenol degradation. Therefore, the microbial community structures in anaerobic degradation of terephthalate and phenol remain to be further characterized. Recently, microbial populations inside wastewater treatment plants have been extensively investigated using 16S rRNA-based culture-independent molecular techniques with an attempt to identify the key players and understand their abundance, dynamics, and distribution within the “black box” in relation to the reactor performance. These in-depth understandings are important to optimize biological processes. 1.3 Objectives The overall objective of this research is to study the microbial community structures responsible for anaerobic terephthalate and phenol degradation using 16S rRNA gene-based molecular approaches. Specific objectives are: Introduction a. To characterize the microbial community structure and dynamics in a thermophilic (55°C) anaerobic laboratory-scale hybrid reactor degrading terephthalate-containing wastewater, b. To study the microbial community structure in a terephthalate-degrading anaerobic hybrid bioreactor operated at 46−50°C, c. To identify the important microbial populations in the methanogenic phenol-degrading enrichments under mesophilic (37°C) and thermophilic (55°C) conditions, and d. To investigate the microbial community structure in a mesophilic full-scale phenol-degrading anaerobic fluidized bed reactor. The results will provide evidence that terephthalate can be degraded under 46−50ºC and 55ºC conditions; improve our understanding of syntrophic microorganisms in the terephthalate and phenol degradation systems; identify microorganisms that are important to the degradation; and provide additional knowledge for future development of methanogenic treatment technologies for terephalate- and phenol-containing wastewaters. 1.4 Organization of the thesis The thesis consists of eight chapters. Chapter provides a general introduction, the problem statements and objectives. Chapter provides a summary of research publications relevant to this study. Chapter describes the experimental materials and methods used. Chapters to describe the results and discussion on the microbial community structures in a thermophilic anaerobic hybrid reactor degrading terephthalate Introduction (Chapter 4), a moderate thermophilic anaerobic hybrid reactor degrading terephthalate (Chapter 5), mesophilic and thermophilic phenol-degrading methanogenic consortia (Chapter 6), and a full-scale phenol-degrading bioreactor (Chapter 7). Last, Chapter provides the overall conclusions of this study and recommendations for future works. 10 References References Altschul, S. F., Madden, T. L., Schaffer, A. A., Zhang, J., Zhang, Z., Miller, W. & Lipman, D. J. (1997). Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25, 3389-3402. Amann, R. I., Ludwig, W. & Schleifer, K. H. (1995). Phylogenetic identification and in situ detection of individual microbial cells without cultivation. Microbiol Rev 59, 143-169. APHA, AWWA & WEF (1998). Standard methods for the examination of water and wastewater, 20th edn. Washington D.C.: American Public Health Association, American Water Works Association, Water Environment Federation. Ashelford, K. E., Chuzhanova, N. A., Fry, J. C., Jones, A. J. & Weightman, A. J. (2005). At least in 20 16S rRNA sequence records currently held in public repositories is estimated to contain substantial anomalies. Appl Environ Microbiol 71, 7724-7736. Autenrieth, R. L., Bonner, J. S., Akgerman, A., Okaygun, M. & McCreary, E. M. (1991). Biodegradation of phenolic wastes. J Hazard Mater 28, 29-53. Becker, J. G., Berardesco, G., Rittmann, B. E. & Stahl, D. A. (2001). Successional changes in an evolving anaerobic chlorophenol-degrading community used to infer relationships between population structure and system-level processes. Appl Environ Microbiol 67, 5705-5714. Borghans, A. J. M. L. & van Driel, A. (1988).Application of the biothane UASB reactor to a chemical wastewater, containing phenol and formaldehyde. In 5th Int Symp Anaerobic Digestion, pp. 627-630. Edited by A. Tilche & A. Rozzi. Bologna, Italy. Boyer, S. L., Flechtner, V. R. & Johansen, J. R. (2001). Is the 16S-23S rRNA internal transcribed spacer region a good tool for use in molecular systematics and population genetics? A case study in cyanobacteria. Mol Biol Evol 18, 1057-1069. 145 References Brauer, S. L., Cadillo-Quiroz, H., Yashiro, E., Yavitt, J. B. & Zinder, S. H. (2006). Isolation of a novel acidiphilic methanogen from an acidic peat bog. Nature 442, 192-194. Bushway, M. H. & Gilman, G. L. (1986).Case history: development of a dedicated sludge application site by Amoco chemicals Co. and the associated permit to operate by South Carolina department of health and environmental control. In 41st Purdue University Industrial Waste Conference, pp. 726-735. Chelsea, MI, USA: Lewis Publishers Inc. Castro, H. F., Williams, N. H. & Ogram, A. (2000). Phylogeny of sulfate-reducing bacteria(1). FEMS Microbiol Ecol 31, 1-9. Chang, Y. J., Nishio, N. & Nagai, S. (1995). Characteristics of granular methanogenic sludge grown on phenol synthetic medium and methanogenic fermentation of phenolic wastewater in a UASB reactor. J Fermentation Bioeng 79, 348-353. Charest, A., Bisaillon, J. G., Lepine, F. & Beaudet, R. (1999). Removal of phenolic compounds from a petrochemical effluent with a methanogenic consortium. Can J Microbiol 45, 235-241. Chen, C. L., Macarie, H., Ramirez, I., Olmos, A., Ong, S. L., Monroy, O. & Liu, W. T. (2004). Microbial community structure in a thermophilic anaerobic hybrid reactor degrading terephthalate. Microbiology 150, 3429-3440. Cheng, S. S., Ho, C. Y. & Wu, J. H. (1997a). Pilot study of UASB process treating PTA manufacturing wastewater. Water Sci Technol 36, 73-82. Cheng, S. S., Wu, J. H., Tseng, I. C. & Hwang, P. C. (1997b).Treatability study of anaerobic fluidized bed bioreactor treating PTA manufacturing wastewater. In Proc Spec Conf on Chemical Process Industries And Environmental Management, pp. 5.1. Cape Town, South Africa. Cole, J. R., Chai, B., Farris, R. J., Wang, Q., Kulam, S. A., McGarrell, D. M., Garrity, G. M. & Tiedje, J. M. (2005). The Ribosomal Database Project (RDP-II): sequences and tools for high-throughput rRNA analysis. Nucleic Acids Res 33, D294-296. 146 References Conrads, G., Citron, D. M., Tyrrell, K. L., Horz, H. P. & Goldstein, E. J. (2005). 16S-23S rRNA gene internal transcribed spacer sequences for analysis of the phylogenetic relationships among species of the genus Porphyromonas. Int J Syst Evol Microbiol 55, 607-613. Cowan, D., Meyer, Q., Stafford, W., Muyanga, S., Cameron, R. & Wittwer, P. (2005). Metagenomic gene discovery: past, present and future. Trends Biotechnol 23, 321-329. Daims, H., Bruhl, A., Amann, R., Schleifer, K. H. & Wagner, M. (1999). The domain-specific probe EUB338 is insufficient for the detection of all Bacteria: development and evaluation of a more comprehensive probe set. Syst Appl Microbiol 22, 434-444. Dojka, M. A., Hugenholtz, P., Haack, S. K. & Pace, N. R. (1998). Microbial diversity in a hydrocarbon- and chlorinated-solvent-contaminated aquifer undergoing intrinsic bioremediation. Appl Environ Microbiol 64, 3869-3877. Dumont, M. G. & Murrell, J. C. (2005). Stable isotope probing - linking microbial identity to function. Nat Rev Microbiol 3, 499-504. Egert, M. & Friedrich, M. W. (2003). Formation of pseudo-terminal restriction fragments, a PCR-related bias affecting terminal restriction fragment length polymorphism analysis of microbial community structure. Appl Environ Microbiol 69, 2555-2562. Embley, T. M., Finlay, B. J., Thomas, R. H. & Dyal, P. L. (1992). The use of rRNA sequences and fluorescent probes to investigate the phylogenetic positions of the anaerobic ciliate Metopus palaeformis and its archaeobacterial endosymbiont. J Gen Microbiol 138, 1479-1487. Evans, W. C. (1977). Biochemistry of the bacterial catabolism of aromatic compounds in anaerobic environments. Nature 270, 17-22. Fajardo, C., Guyot, J.-P., Macarie, H. & Monroy, O. (1997). Inhibition of anaerobic digestion by terephthalic acid and its aromatic by products. Water Sci Technol 36, 83–90. 147 References Fang, H. H., Liang, D. W., Zhang, T. & Liu, Y. (2006). Anaerobic treatment of phenol in wastewater under thermophilic condition. Water Res 40, 427-434. Fang, H. H. P., Chui, H. K., Li, Y. Y. & Chen, T. (1994). Performance and granule characteristics of UASB process treating wastewater with hydrolyzed proteins. Wat Sci Technol 30, 55-63. Fang, H. H. P., Chen, T., Li, Y. Y. & Chui, H. K. (1996). Degradation of phenol in wastewater in an upflow anaerobic sludge blanket reactor. Water Res 30, 1353-1360. Fang, H. H. P. & Zhou, G. M. (2000). Degradation of phenol and p-cresol in reactors. Water Sci Technol 42, 237-244. Fang, H. H. P., Liu, Y., Ke, S. Z. & Zhang, T. (2004). Anaerobic degradation of phenol in wastewater at ambient temperature. Water Sci Technol 49, 95-102. Ferris, M. J., Muyzer, G. & Ward, D. M. (1996). Denaturing gradient gel electrophoresis profiles of 16S rRNA-defined populations inhabiting a hot spring microbial mat community. Appl Environ Microbiol 62, 340-346. Fligg, J. E. (1996). Polyester growth and its impact on PTA study. Int Fiber J 11, 16-22. Franck, H. G. & Stadelhofer, L. W. (1988). p-Xylene and its derivatives: terephthalic acid. In Industrial Aromatic Chemistry (Chapter 7), pp. 283-290. Edited by H. G. Franck. New York, NY: Springer-Verlag. Gallert, C., Knoll, G. & Winter, J. (1991). Anaerobic carboxylation of phenol to benzoate: use of deuterated phenols revealed carboxylation exclusively in the C4-position. Appl Microbiol Biotechnol 36, 124-129. Gich, F., Garcia-Gil, J. & Overmann, J. (2001). Previously unknown and phylogenetically diverse members of the green nonsulfur bacteria are indigenous to freshwater lakes. Arch Microbiol 177, 1-10. Guiot, S. R., Pauss, A. & Costerton, J. W. (1992). A structured model of the anaerobic granule consortium. Wat Sci Technol 25, 1-10. 148 References Hall, T. A. (1999). BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucl Acids Symp Ser 41, 95-98. Harwood, C. S., Burchhardt, G., Herrmann, H. & Fuchs, G. (1999). Anaerobic metabolism of aromatic compounds via the benzoyl-CoA pathway. FEMS Microbiol Rev 22, 439-458. Heider, J. & Fuchs, G. (1997). Anaerobic metabolism of aromatic compounds. Eur J Biochem 243, 577-596. Heuer, H., Krsek, M., Baker, P., Smalla, K. & Wellington, E. M. (1997). Analysis of actinomycete communities by specific amplification of genes encoding 16S rRNA and gel-electrophoretic separation in denaturing gradients. Appl Environ Microbiol 63, 3233-3241. Huber, T., Faulkner, G. & Hugenholtz, P. (2004). Bellerophon: a program to detect chimeric sequences in multiple sequence alignments. Bioinformatics 20, 2317-2319. Hugenholtz, P., Pitulle, C., Hershberger, K. L. & Pace, N. R. (1998). Novel division level bacterial diversity in a Yellowstone hot spring. J Bacteriol 180, 366-376. Hugenholtzt, P. & Huber, T. (2003). Chimeric 16S rDNA sequences of diverse origin are accumulating in the public databases. Int J Syst Evol Microbiol 53, 289-293. Hulshoff Pol, L. W., de Castro Lopes, S. I., Lettinga, G. & Lens, P. N. (2004). Anaerobic sludge granulation. Water Res 38, 1376-1389. Imachi, H., Sekiguchi, Y., Kamagata, Y., Hanada, S., Ohashi, A. & Harada, H. (2002). Pelotomaculum thermopropionicum gen. nov., sp. nov., an anaerobic, thermophilic, syntrophic propionate-oxidizing bacterium. Int J Syst Evol Microbiol 52, 1729-1735. Imachi, H., Sekiguchi, Y., Kamagata, Y. & other authors (2006). Non-sulfate-reducing, syntrophic bacteria affiliated with desulfotomaculum cluster I are widely distributed in methanogenic environments. Appl Environ Microbiol 72, 2080-2091. 149 References Janse, I., Meima, M., Kardinaal, W. E. & Zwart, G. (2003). High-resolution differentiation of Cyanobacteria by using rRNA-internal transcribed spacer denaturing gradient gel electrophoresis. Appl Environ Microbiol 69, 6634-6643. Juteau, P., Cote, V., Duckett, M. F., Beaudet, R., Lepine, F., Villemur, R. & Bisaillon, J. G. (2005). Cryptanaerobacter phenolicus gen. nov., sp. nov., an anaerobe that transforms phenol into benzoate via 4-hydroxybenzoate. Int J Syst Evol Microbiol 55, 245-250. Kamagata, Y. & Tamaki, H. (2005). Cultuvation of uncultured fastidious microbes. Microbes Environ 2005, 85-91. Kane, M. D., Poulsen, L. K. & Stahl, D. A. (1993). Monitoring the enrichment and isolation of sulfate-reducing bacteria by using oligonucleotide hybridization probes designed from environmentally derived 16S rRNA sequences. Appl Environ Microbiol 59, 682-686. Karlsson, A., Ejlertsson, J., Nezirevic, D. & Svensson, B. H. (1999). Degradation of phenol under meso- and thermophilic, anaerobic conditions. Anaerobe 5, 25-35. Kengen, S. W., Breidenbach, C. G., Felske, A., Stams, A. J., Schraa, G. & de Vos, W. M. (1999). Reductive dechlorination of tetrachloroethene to cis-1, 2-dichloroethene by a thermophilic anaerobic enrichment culture. Appl Environ Microbiol 65, 2312-2316. Khan, K. A., Suidan, M. T. & Cross, W. H. (1981). Anaerobic activated carbon filter for the treatment of phenol-bearing wastewater. J Water Pollut Control Fed 53, 1519-1532. Kim, B. R., Chian, E. S. K., Cross, W. H. & Cheng, S. S. (1986). Adsorption, desorption, and biodegradation in an anaerobic, granular activated carbon reactor for the removal of phenol. J WPCF 58, 35-40. Kirk-Othmer (1978). Encyclopedia of Chemical Technology, 3rd edn. New York: Wiley-Interscience. 150 References Kleerebezem, R., Mortier, J., Hulshoff Pol, L. W. & Lettinga, G. (1997). Anaerobic pretreatment of petrochemical effluents: terephthalic acid wastewater. Water Sci Technol 36, 237-248. Kleerebezem, R., Hulshoff Pol, L. W. & Lettinga, G. (1999a). Anaerobic degradation of phthalate isomers by methanogenic consortia. Appl Environ Microbiol 65, 1152-1160. Kleerebezem, R., Hulshoff Pol, L. W. & Lettinga, G. (1999b). The role of benzoate in anaerobic degradation of terephthalate. Appl Environ Microbiol 65, 1161-1167. Kleerebezem, R., Ivalo, M., Hulshoff Pol, L. W. & Lettinga, G. (1999c). High-rate treatment of terephthalate in anaerobic hybrid reactors. Biotechnol Prog 15, 347-357. Kleerebezem, R. & Macarie, H. (2003).Treating industrial wastewater: anaerobic digestion comes of age. In Chemical Engineering, April, pp. 56-64. Kleerebezem, R., Beckers, J., Hulshoff Pol, L. W. & Lettinga, G. (2005). High rate treatment of terephthalic acid production wastewater in a two-stage anaerobic bioreactor. Biotechnol Bioeng 91, 169-179. Kobayashi, T., Hashinaga, T., Mikami, E. & Suzuki, T. (1989). Methanogenic degradation of phenol and benzoate in acclimated sludge. Water Sci Technol 21, 55-65. Kumar, S., Tamura, K. & Nei, M. (2004). MEGA3: Integrated software for Molecular Evolutionary Genetics Analysis and sequence alignment. Brief Bioinform 5, 150-163. Lain, D. M., Hwang, S. U., Chau, L. T. & Cheng, S. S. (1999).Patent number: 105045. Taiwan, ROC. Lao, S. G. (2002). Mechanisms of granular activated carbon anaerobic fluidized-bed process for treating phenols wastewater. J Environ Sci (China) 14, 132-135. Lau, C. M. (1977).Staging aeration for high-efficiency treatment of aromatics plant wastewater. In Proc Ind Waste Conf, pp. 63-74. Purdue Univ. 151 References Lettinga, G. (1995). Anaerobic digestion and wastewater treatment systems. Antonie Van Leeuwenhoek 67, 3-28. Liu, W.-T., Huang, C. L., Hu, J. Y., Song, L. F., Ong, S. L. & Ng, W. J. (2002). Denaturing gradient gel electrophoresis polymorphism for rapid 16S rDNA clone screening and microbial diversity estimation. J Biosci Bioeng 93. Liu, W. T., Marsh, T. L., Cheng, H. & Forney, L. J. (1997). Characterization of microbial diversity by determining terminal restriction fragment length polymorphisms of genes encoding 16S rRNA. Appl Environ Microbiol 63, 4516-4522. Liu, Y. & Tay, J. H. (2002). The essential role of hydrodynamic shear force in the formation of biofilm and granular sludge. Water Res 36, 1653-1665. Liu, Y. & Tay, J. H. (2004). State of the art of biogranulation technology for wastewater treatment. Biotechnol Adv 22, 533-563. Lomans, B. P., Maas, R., Luderer, R., Op den Camp, H. J., Pol, A., van der Drift, C. & Vogels, G. D. (1999). Isolation and characterization of Methanomethylovorans hollandica gen. nov., sp. nov., isolated from freshwater sediment, a methylotrophic methanogen able to grow on dimethyl sulfide and methanethiol. Appl Environ Microbiol 65, 3641-3650. Londry, K. L. & Fedorak, P. M. (1991). Benzoic acid intermediates in the anaerobic biodegradation of phenols. Can J Microbiol 38, 1-11. Loy, A., Lehner, A., Lee, N., Adamczyk, J., Meier, H., Ernst, J., Schleifer, K. H. & Wagner, M. (2002). Oligonucleotide microarray for 16S rRNA gene-based detection of all recognized lineages of sulfate-reducing prokaryotes in the environment. Appl Environ Microbiol 68, 5064-5081. Ludwig, W., Strunk, O., Westram, R. & other authors (2004). ARB: a software environment for sequence data. Nucleic Acids Res 32, 1363-1371. Macarie, H. & Guyot, J. P. (1992). Inhibition of the methanogenic fermentation of p-toluic acid (4-methylbenzoic acid) by acetate. Appl Microbiol Biotechnol 38, 398-402. 152 References Macarie, H., Noyola, A. & Guyot, J.-P. (1992). Anaerobic treatment of a petrochemical wastewater from a terephthalic acid plant. Water Sci Technol 25, 223-235. Macarie, H. (2000). Overview of the application of anaerobic treatment to chemical and petrochemical wastewaters. Water Sci Technol 42, 201-214. MacLeod, F. A., Guiot, S. R. & Costerton, J. W. (1990). Layered structure of bacterial aggregates produced in an upflow anaerobic sludge bed and filter reactor. Appl Environ Microbiol 56, 1598-1607. Madigan, M. T. & Martinko, J. M. (2006). Brock Biology of Microorganisms, Prentice-Hall, Upper Saddle River, New Jersey. Madsen, E. L. (2006). The use of stable isotope probing techniques in bioreactor and field studies on bioremediation. Curr Opin Biotechnol 17, 92-97. Nakhla, G. F. & Suidan, M. T. (1995). Anaerobic toxic wastes treatment: diluation effects. J Hazard Mater 42, 71-86. Okabe, S., Kindaichi, T. & Ito, T. (2004). MAR-FISH-- An ecophysiological approach to link phylogenetic affiliation and in situ metabolic activity of microorganisms at a single-cell resolution. Microbes Environ 19, 83-98. Park, C.-M. & Sheehan, R. J. (1996). Phthalic acids and other benzenepolycarboxylic acids. In Kirk-Othmer Encyclopedia of Chemical Technology, pp. 991-1043. Edited by R. E. Kirk, D. F. Othmer, J. Kroschwitz & M. Howe-Grant. New York, USA: John Wiley and Sons. Patterson, J. W. (1975). Treatment technology for phenols in wastewater treatment technology. Ann Arbor, MI: Ann Arbor Science Publishers. Qiu, Y. L., Sekiguchi, Y., Imachi, H., Kamagata, Y., Tseng, I. C., Cheng, S. S., Ohashi, A. & Harada, H. (2004). Identification and isolation of anaerobic, syntrophic phthalate isomer-degrading microbes from methanogenic sludges treating wastewater from terephthalate manufacturing. Appl Environ Microbiol 70, 1617-1626. 153 References Qiu, Y. L., Sekiguchi, Y., Hanada, S., Imachi, H., Tseng, I. C., Cheng, S. S., Ohashi, A., Harada, H. & Kamagata, Y. (2006). Pelotomaculum terephthalicum sp. nov. and Pelotomaculum isophthalicum sp. nov.: two anaerobic bacteria that degrade phthalate isomers in syntrophic association with hydrogenotrophic methanogens. Arch Microbiol 185, 172-182. Raskin, L., Stromley, J. M., Rittmann, B. E. & Stahl, D. A. (1994). Group-specific 16S rRNA hybridization probes to describe natural communities of methanogens. Appl Environ Microbiol 60, 1232-1240. Razo-Flores, E., Iniestra-Gonzalez, M., Field, J. A., Olguin-Lora, P. & Puig-Grajales, L. (2003). Biodegradation of mixtures of phenolic compounds in an upward-flow anaerobic sludge blanket reactor. J Environ Eng 129, 999-1006. Razo-Flores, E., Macarie, H. & Morier, F. (2006). Application of biological treatment systems for chemical and petrochemical wastewaters. In Advanced Biological Treatment Processes for Industrial Wastewaters, pp. 267-297. Edited by F. J. Cervantes, S. G. Pavlostathis & A. C. van Haandel. London, UK: IWA Publishing. Savostianoff, D. & Didier, S. N. (1993). DMT_PTA, Asia advances towards an arushing domination. Inf Chim 352, 119-129. Schink, B. (1997). Energetics of syntrophic cooperation in methanogenic degradation. Microbiol Mol Biol Rev 61, 262-280. Schlotelburg, C., von Wintzingerode, F., Hauck, R., Hegemann, W. & Gobel, U. B. (2000). Bacteria of an anaerobic 1,2-dichloropropane-dechlorinating mixed culture are phylogenetically related to those of other anaerobic dechlorinating consortia. Int J Syst Evol Microbiol 50 Pt 4, 1505-1511. Schlötelburg, C. (2001).Mikrobielle Diversitat und Dynamik einer 1,2-Dichlorpropan dechlorierenden Mischkultur. Berlin: Humboldt-University. Sekiguchi, Y., Kamagata, Y., Syutsubo, K., Ohashi, A., Harada, H. & Nakamura, K. (1998). Phylogenetic diversity of mesophilic and thermophilic granular sludges determined by 16S rRNA gene analysis. Microbiology 144 (Pt 9), 2655-2665. 154 References Sekiguchi, Y., Kamagata, Y., Nakamura, K., Ohashi, A. & Harada, H. (1999). Fluorescence in situ hybridization using 16S rRNA-targeted oligonucleotides reveals localization of methanogens and selected uncultured bacteria in mesophilic and thermophilic sludge granules. Appl Environ Microbiol 65, 1280-1288. Sekiguchi, Y., Kamagata, Y. & Harada, H. (2001a). Recent advances in methane fermentation technology. Curr Opin Biotechnol 12, 277-282. Sekiguchi, Y., Takahashi, H., Kamagata, Y., Ohashi, A. & Harada, H. (2001b). In situ detection, isolation, and physiological properties of a thin filamentous microorganism abundant in methanogenic granular sludges: a novel isolate affiliated with a clone cluster, the green non-sulfur bacteria, subdivision I. Appl Environ Microbiol 67, 5740-5749. Sekiguchi, Y. (2006). Yet-to-be cultured microorganisms relevant to methane fermentation processes. Microbes Environ 21, 1-15. Shelley, S. (1991). And the winner is. Amoco chemical. Chem Eng 98, 90-93. Stahl, D. A. & Amann, R. (1991). Development and application of nucleic acid probes. In Nucleic Acid Techniques in Bacterial Systematics, pp. 205-248. Edited by E. Stackebrandt & M. Goodfellow. New York: Wiley. Stickler, D. (1999). Biofilms. Curr Opin Microbiol 2, 270-275. Streit, W. R. & Schmitz, R. A. (2004). Metagenomics--the key to the uncultured microbes. Curr Opin Microbiol 7, 492-498. Stubner, S. & Meuser, K. (2000). Detection of Desulfotomaculum in an Italian rice paddy soil by 16S ribosomal nucleic acid analyses. FEMS Microbiol Ecol 34, 73-80. Suidan, M. T., Cross, W. H., Fong, M. & Calvert, J. W. (1981). Anaerobic carbon filter for degradation of phenols. J Environ Eng, ASCE 107 (EE3), 563-579. Suidan, M. T., Siekerka, G. L., Kao, S. W. & Pfeffer, J. T. (1983a). Anaerobic filters for the treatment of coal gasification wastewater. Biotechnol Bioeng XXV, 1581-1596. 155 References Suidan, M. T., Strubler, C. E., Kao, S. W. & Pfeffer, J. T. (1983b). Treatment of coal gasification wastewater with anaerobic filter technology. J WPCF 55, 1263-1270. Suresh, B., Davenport, B. & Mavar, L. (2001).Dimethyl terephthalate (DMT) and terephthalic acid (TPA). In SRI International's Chemical Economics Handbook Program (http://cehsricsricom/Public/Reports/). Suzanne, S. (1991). Winner is AMOCO chemical. Chemical Engineering, 90-93. Syutsubo, K., Harada, H., Ohashi, A. & Suzuki, H. (1997). An effective start-up of thermophilic UASB reactor by seeding mesophilically-grown granular sludge. Wat Sci Technol 36, 391-398. Tay, J. H., He, Y. X. & Yan, Y. G. (2000). Anaerobic biodegranulation using phenol as the sole carbon source. Water Environ Res 72, 189-194. Tay, J. H., He, Y. X. & Yan, Y. G. (2001). Improved anaerobic degradation of phenol with supplemental glucose. J Environ Eng, ASCE 127, 38-45. Thierry, S., Ramirez, I., Allouche, C., Ferrer, H. & Macarie, H. (1999).Thermophilic anaerobic treatment of purified terephthalic acid wastewater. In Proc Int Conf Wast Min End of Pipe Treat Chem Petrochem Ind. Merida, Yucatán, Mexico. Thompson, J. R., Marcelino, L. A. & Polz, M. F. (2002). Heteroduplexes in mixed-template amplifications: formation, consequence and elimination by 'reconditioning PCR'. Nucleic Acids Res 30, 2083-2088. Tiwari, M. K., Guha, S., Harendranath, C. S. & Tripathi, S. (2006). Influence of extrinsic factors on granulation in UASB reactor. Appl Microbiol Biotechnol 71, 145-154. Tsuno, H., Kawamura, M. & Oya, T. (2006). Application of biological activated carbon anaerobic reactor for treatment of hazardous chemicals. Water Sci Technol 53, 251-260. Tyson, G. W., Chapman, J., Hugenholtz, P. & other authors (2004). Community structure and metabolism through reconstruction of microbial genomes from the environment. Nature 428, 37-43. 156 References Uemura, S. & Harada, H. (1993). Microbial characteristics of methanogenetic sludge consortia developed in thermophilic UASB reactors. Appl Microbiol Biotechnol 39, 654-660. Urakawa, H., Noble, P. A., El Fantroussi, S., Kelly, J. J. & Stahl, D. A. (2002). Single-base-pair discrimination of terminal mismatches by using oligonucleotide microarrays and neural network analyses. Appl Environ Microbiol 68, 235-244. van Lier, J. B. (1996). Limitations of thermophilic anaerobic wastewater treatment and the consequences for process design. Antonie Van Leeuwenhoek 69, 1-14. van Lier, J. B., Rebac, S. & Lettinga, G. (1997). High-rate anaerobic wastewater treatment under psychrophilic and thermophilic conditions. Water Sci Technol 35, 199-206. Vanduffel, J. M. (1993).Anaerobic treatment of organic acids. In National Conference on Anaerobic Treatment of Complex Wastewaters. Breda, The Netherlands. Veeresh, G. S., Kumar, P. & Mehrotra, I. (2005). Treatment of phenol and cresols in upflow anaerobic sludge blanket (UASB) process: a review. Water Res 39, 154-170. von Wintzingerode, F., Selent, B., Hegemann, W. & Gobel, U. B. (1999). Phylogenetic analysis of an anaerobic, trichlorobenzene-transforming microbial consortium. Appl Environ Microbiol 65, 283-286. Wagner, M. (2004). Deciphering functions of uncultured microorganisms. ASM News 70, 63-70. Wallner, G., Amann, R. & Beisker, W. (1993). Optimizing fluorescent in situ hybridization with rRNA-targeted oligonucleotide probes for flow cytometric identification of microorganisms. Cytometry 14, 136-143. Wang, Y. T., Suidan, M. T. & Rittmann, B. E. (1986). Anaerobic treatment of phenol by an expanded-bed reactor. J Wat Poll Control Fed 58, 227-233. 157 References Weisburg, W. G., Barns, S. M., Pelletier, D. A. & Lane, D. J. (1991). 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol 173, 697-703. Wu, J. H., Liu, W. T., Tseng, I. C. & Cheng, S. S. (2001). Characterization of microbial consortia in a terephthalate-degrading anaerobic granular sludge system. Microbiology 147, 373-382. Yamada, T., Sekiguchi, Y., Imachi, H., Kamagata, Y., Ohashi, A. & Harada, H. (2005). Diversity, localization, and physiological properties of filamentous microbes belonging to Chloroflexi subphylum I in mesophilic and thermophilic methanogenic sludge granules. Appl Environ Microbiol 71, 7493-7503. Young, L. Y. & Rivera, M. D. (1985). Methanogenic degradation of four phenolic componds. Water Res 19, 1325-1332. Zhang, T., Ke, S. Z., Liu, Y. & Fang, H. P. (2005). Microbial characteristics of a methanogenic phenol-degrading sludge. Water Sci Technol 52, 73-78. Zhou, G. M. & Fang, H. H. P. (1997). Co-degradation of phenol and m-cresol in an UASB reactor. Bioresource Technol 61, 47-52. 158 Publications Publications Journal papers: Chen, C.-L., Wu, J.-H., Tseng, I-C., Liang, T.-M. & Liu, W.-T. (2008). Diversity and localization of microbial consortium in a full-scale anaerobic reactor treating phenolic wastewater. Microbes and Environments (submitted). Chen, C.-L., Wu, J.-H. & Liu, W.-T. (2008). Identification of important microbial populations in the mesophilic and thermophilic phenol-degrading methanogenic consortia. Water Research 42, 1963-1976. Chen, C.-L., Macarie, H., Ramirez, I., Olmos, A., Ong, S. L., Monroy, O. & Liu, W.-T. (2004). Microbial community structure in a thermophilic anaerobic hybrid reactor degrading terephthalate. Microbiology 150, 3429-3440. Chen, C.-L., Liu, W.-T., Chong, M. L., Wong, M. T., Ong, S. L., Seah, H. & Ng, W. J. (2004). Community structure of microbial biofilms associated with membrane-based water purification processes as revealed using a polyphasic approach. Applied Microbiology and Biothechnology 63, 466-473. Conference papers: Chen, C.-L. & Liu, W.-T. (2008). Diversity and localization of microbial consortium in a full-scale phenol-degrading granular activated carbon anaerobic reactor. Biofilm Technologies, Singapore, 8-10 January. Chen, C.-L., Ong, S. L. & Liu, W.-T. (2004). Microbial diversity in anaerobic degradation of phthalate isomers under thermophilic conditions. 13th Kaist-Kyoto-NTU-NUS joint symposium on environmental engineering, Singapore, 17-18 June. 159 Publications Chen, C.-L., Ong, S. L. & Liu, W.-T. (2003). Characterization of microbial community in a thermophilic terephthalate-degrading anaerobic sludge system. Fourth Combined Scientific Meeting (Incorating Second Singapore Microarry Meeting), Singapore, 15-18 January. Liu, W.-T., Chen, C.-L., Chong, M. L., Ong, S. L. & Ng, W. J. (2002). Microbial community structure of membrane biofouling in MF- and RO-based water purification processes as determined by molecular techniques. 13th Annual Meeting of the North American Membrane Society (NAMS), Long Beach, California, USA, 11-15 May. 160 [...]... (ºC) Reactor volume (L) Full − 1280 m3 Lab 37 0.66 Lab 37 2.8 Lab 37 2.8 Lab 37 2.8 Sucrose ( 134 ) 0 .33 0.5 Phenol/ p -cresol: 800 /30 0 − Lab 35 2.0 0.5 37 2.0 Lab 30 0.16 Lab 26 2.8 Phenol: 1260 Phenol: 1260 Phenol/ p -cresol: 280/ 132 Phenol: 1260 6 Lab Glucose (260) Glucose (120) Acetate (55) Glucose (−) 0.5 1 0.5 0.50.6 0.5 8 7 6 Phenol: 98±1 Phenol: 100 p-cresol: 93 Phenol: 98 References (Borghans & van... matters and the standard Gibbs free-energy Fermentative reaction ∆Go' (kJ/reaction) Terephthalate + 8 H2O → 3 acetate- + 2 HCO3- + 3 H+ + 3 H2 +38 .9 Phenol + 5 H2O → 3 acetate- + 3 H+ + 2 H2 +10.2 Hydrogenotrophic methanogenesis 4 H2 + HCO3- + H+ → CH4 + 3 H2O − 131 .2 Acetotrophic methanogensis Acetate- + H2O → CH4 + HCO3- 2.2.2 32 .6 Proposed terephthalate degradation pathway Anaerobic degradation of terephthalate. .. terms “phenols”, “total phenols” or “phenolics” used for wastewaters or contaminated sites denote simple phenol or a mixture of phenolic compounds, such as phenol, cresol isomers (ortho-, meta- and para-), resorcinol, hydriquinone and dimethyl phenol Phenols are often found in wastewaters generated from coal gasification, coke ovens, and petroleum-related manufacturing processes (e.g., phenolic resin,... studies Using microscopy, Kleerebezem and co-workers (1999a, b) observed that spore-forming rods (3 to 4 µm) were associated with Methanosaeta- and Methanospirillum-like cells in a terephthalate- degrading consortium degrading bacteria was 7−28 days The doubling time of these terephthalate- Wu and co-workers (2001) studied the microbial community structure in a mesophilic terephthalate- degrading granular... dominant microbial populations could be identified However, they observed that the microbial populations under thermophilic conditions were different from those observed in the bioreactor operated under ambient temperature The microbial communities involving in thermophilic phenol degradation remain to be characterized 32 Materials and Methods Chapter 3 Materials and Methods 33 Materials and Methods 3. 1... extremely slow in the media containg isophthalate in the presence of Methanospirillum hungatei cells A defined “pure co-culture” of strain JIT was obtained by co-cultured with M hungatei with 3- hydroxybenzoate (5 mM) and yeast extract (0.02%) as the carbon substrate The cells are 0.8−1.0 µm wide and 2.0 3. 0 µm long, occurring singly or in pairs The spores are spherical and at central of the cell Tiny colonies,... 2.1 Anaerobic treatment of terephthalate- and phenol- containing wastewaters During the past 20 years, anaerobic digestion processes have become an attractive biological approach to treat toxic and poorly biodegradable wastewaters (e.g., petrochemical wastewaters) (Lettinga, 1995) In these processes, organic compounds are degraded and converted into methane and carbon dioxide in the absence of oxygen and. .. populations in this UASB system Like anaerobic degradation of terephthalate, the initial conversion of phenol has been identified as the rate-limiting step As a result, non-layered structure of phenol- degrading granules was observed in the UASB reactors operated at 26°C (Fang et al., 1996; Zhang et al., 2005) 31 Literature Review Fang and co-workers (2006) also enriched a phenol- degrading methanogenic... with that reported by Evans (1977) Thus, the degradation pathway of phenol via caproate needs to be further confirmed 2 .3 Microbial communities for terephthalate and phenol degradation 2 .3. 1 Microorganisms involved in terephthalate degradation 2 .3. 1.1 Culture-dependent studies Two mesophilic terephthalate- degrading bacteria, Pelotomaculum terephthalicum and Pelotomaculum isophthalicum have been isolated... and Methods 3. 1 Terephthalate- and phenol- degrading microbial consortia 3. 1.1 Thermophilic anaerobic terephthalate- degrading reactor A 1.2-liter laboratory-scale hybrid reactor, packed with 69 polypropylene Pall rings (Flexiring®, Koch Inc.) (total volume = 400 mL), was used to enrich anaerobic microbial consortia degrading terephthalate under thermophilic conditions (Fig 3. 1) was inoculated with seed . represent as the main population involved in terephthalate or phenol degradation. Therefore, the microbial community structures in anaerobic degradation of terephthalate and phenol remain to be further. coke ovens and oil refining processes. The concentration of phenol and phenolic compounds in wastewaters is approximately 1.0−1.7 x 10 4 mg/L, and the COD contributed by phenol and phenolic. development of thermophilic techniques to treat phenol- containing wastewater is relatively limited. A better understanding of the phenol- degrading microbial community could provide crucial information