Abstract
The strictly anaerobic, pentachlorophenol (PCP) degrading bacterium DCB-2 was immobilized in an Upflow Anaerobic Sludge Blanket (UASB) reactor containing sterile granules. PCP and lactate were fed to the reactor and the concentration of chlorophenols in the effluent were monitored for 641 days. PCP was found to be degraded and transformed into 3.4.5-trichlorophenol in the reactor where DCB-2 was introduced into the granular sludge. PCP was still transformed to 3.4.5-trichlorophenol when the hydraulic retention time was decreased to six hours which was much lower than the generation time of DCB-2 insuring no free living cells in the reactor. This indicated that DCB-2 was immobilized in the granular layer. A control reactor that contained only sterile granules did not dechlorinate PCP indicating that the performance in the inoculated reactor was only due to the introduced bacteria. Immobilization of DCB-2 in the granules was further demonstrated by adding an antibody raised against DCB-2 to sliced granules. Bacteria thus visualized formed a net structure inside the granules. No DCB-2 bacteria could be found in granules from the control reactor. When lactate was omitted from the influent, the reactor still dechlorinated PCP in accordance with our findings that lactate was not used by DCB-2. This suggested that the reducing equivalents for reductive dechlorination were derived from the granules themselves. The reactor performance was 120 μmol·l reactor-1·day-1, comparable to the best described performance of a UASB-reactor and to aerobic reactors. Our study demonstrates that granules can be constructed which possess specific abilities such as a dechlorinating activity and at the same time be high performing. This result have implications for eco-engineering of granules for anaerobic treatment of contaminated waters.
Similar content being viewed by others
References
Ahring BK, Christiansen N, Mathrani I, Hendriksen HV, Macario AJL & Conway de Macario E (1992) Introduction of a de novo bioremediation ability, aryl reductive dechlorination, into anaerobic granular sludge by inoculation of sludge withDesulfomonile tiedjei. Appl. Environ. Microbiol. 58: 3677–3682
American Public Health Association, Washington, D.C. (1985) Standard methods for the examination of water and wastewater
Angelidaki I, Petersen SP & Ahring BK (1990) Effects of lipids on thermophilic anaerobic digestion and reduction of lipid inhibition upon addition of bentonite. Appl. Microbiol. Biotechnol. 33: 469–472
BabcockJr. RW, Ro KS, Hsieh C-C & Stenstrom MK (1992) Development of an off-line enricher-reactor process for activated sludge degradation of hazardous wastes. Wat. Environ. Res. 64: 782–791
Briglia M, Middeldorp PJM & Salkinoja-Salonen MS (1994) Mineralization performance ofRhodococcus chlorophenolicus strain PCP-1 in contaminated soil simulating on site conditions. Soil. Biol. Biochem. 26: 377–385
Comeau Y, Greer CW & Samson R (1993) Role of inoculum preparation and density on the bioremediation of 2,4-D-contaminated soil by bioaugmentation. Appl. Microbiol. Biotechnol. 38: 681–687
DeWeerd KA, Mandelco L, Tanner RS, Woese CR & Suflita JM (1990)Desulfomonile tiedjei gen. nov. and sp. nov., a novel anaerobic, dehalogenating, sulfate-reducing bacterium. Arch. Microbiol. 154: 23–30
Dolfing J & Tiedje JM (1986) Hydrogen cycling in a three-tiered food web growing on the methanogenic conversion of 3-chlorobenzoate. FEMS Microbiol. Ecol. 38: 293–298
Edgehill RU & Finn RF (1983a) Activated sludge treatment of synthetic wastewater containing pentachlorophenol. Biotechnol. Bioeng. 25: 2165–2176.
Edgehill RU & Finn RK (1983b) Microbial treatment of soil to remove pentachlorophenol. Appl. Environ. Microbiol. 45: 1122–1125
German Collection of Microorganisms and Cell Cultures (DSM) (1989) Catalogue of strains. DSM — German Collection of Microorganisms and Cell Cultures, Braunschweig
Hendriksen HV & Ahring BK (1993) Anaerobic dechlorination of pentachlorophenol in fixed-film and upflow anaerobic sludge blanket reactors using different inocula. Biodegrad. 3: 399–408
Hendriksen HV, Larsen S & Ahring BK (1992a) Influence of a supplemental carbon source on anaerobic dechlorination of pentachlorophenol in granular sludge. Appl. Environ. Microbiol. 58: 365–370
Hendriksen HV, Larsen S & Ahring BK (1992b) Errata. Influence of a supplemental carbon source on anaerobic dechlorination of pentachlorophenol in granular sludge. Appl. Environ. Microbiol. 58: 1073
Hwang P-C & Cheng S-S (1991) The influence of glucose supplement on the degradation of catechol. Wat. Sci. Tech. 23: 1201–1209
Krumme ML, Smith RL, Egestorff J, Thiem SM, Tiedje JM, Timmis KN & Dwyer DF (1985) Behaviour of pollutant-degrading microorganisms in aquifers: Predictions for genetically engineered organisms. Environ. Sci. Technol. 28: 1134–1138
Larsen S, Hendriksen HV & Ahring BK (1991) Potential for thermophilic (50°C) anaerobic dechlorination of pentachlorophenol in different ecosystems. Appl. Environ. Microbiol. 57: 2085–2090
Macario AJL & Conway de Macario E (1983) Antigenic fingerprinting of methanogenic bacteria with polyclonal antibody probes. Syst. Appl. Microbiol. 4: 451–458
Macario AJL & Conway de Macario E (1985) Monoclonal antibodies of predefined molecular specificity for identification and classification of methanogens and for probing their ecological niches. In: Macario AJL, Conway de Macario E (Eds) Monoclonal antibodies against bacteria, pp 213–247. Academic Press, Inc., Orlando, Florida
Madsen T & Licht D (1992) Isolation and characterization of an anaerobic chlorophenol-transforming bacterium. Appl. Environ. Microbiol. 58: 2874–2878
Mikesell MD & Boyd SA (1986) Complete reductive dechlorination and mineralization of pentachlorophenol by anaerobic microorganisms. Appl. Environ. Microbiol. 52: 861–865
Mikesell MD & Boyd SA (1988) Enhancement of pentachlorophenol degradation in soil through induced anaerobiosis and bioaugmentation with anaerobic sewage sludge. Environ. Sci. Technol. 22: 1411–1414.
Nicholson DK, Woods SL, Istok JD & Peek DC (1992) Reductive dechlorination of chlorophenols by a pentachlorophenol-acclimated methanogenic consortium. Appl. Environ. Microbiol. 58: 2280–2286
Puhakka JA & Järvinen K (1992) Aerobic fluidized-bed treatment of polychlorinated phenolic wood preservative constituents. Wat. Res. 26: 765–770
Shelton DR & Tiedje JM (1984) Isolation and partial characterization of bacteria in an anaerobic consortium that mineralizes 3-chlorobenzoic acid. Appl. Environ. Microbiol. 48: 23–30
Sørensen AH & Winther-Nielsen M, Ahring BK (1991) Kinetics of lactate, acetate and propionate in unadapted and lactate-adapted thermophilic, anaerobic sewage sludge: the influence of sludge adaptation for start-up of thermophilic UASB-reactors. Appl. Microbiol. Biotechnol. 34: 823–827
Utkin I, Woese C & Wiegel J (1994) Isolation and characterization ofDesulfitobacterium dehalogenans gen. nov., sp. nov., an anaerobic bacterium which reductively dechlorinates chlorophenolic compounds. Int. J. Syst. Bacteriol. 44: 612–619
Woods SL, Ferguson JF & Benjamin MM (1989) Characterization of chlorophenol and chloromethoxybenzene biodegradation during anaerobic treatment. Environ. Sci. Technol. 23: 62–68
Wu W-M, Bhatnager L & Zeikus JG (1993) Performance of anaerobic granules for degradation of pentachlorophenol. Appl. Environ. Microbiol. 59: 389–397
Zehnder ABJ & Wuhrmann K (1976) Titanium(III)-citrate as a nontoxic oxidation-reduction buffering system for the culture of obligate anaerobes. Science 194: 1165–1166
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Christiansen, N., Ahring, B.K. Introduction of ade novo bioremediation activity into anaerobic granular sludge using the dechlorinating bacterium DCB-2. Antonie van Leeuwenhoek 69, 61–66 (1996). https://doi.org/10.1007/BF00641612
Issue Date:
DOI: https://doi.org/10.1007/BF00641612