Abstract
Molecular approaches based on both whole-cell and extracted DNA were applied to assess chronic and acute effects of copper on the ammonia oxidizing bacteria (AOB) community in an activated sludge system. The ammonia monooxygenase amoA gene was chosen as the functional marker to evaluate changes in the AOB community. Using in situ polymerase chain reaction, we were able to visualize the peripheric distribution of the amoA gene-possessing bacteria in activated sludge flocs. The AOB biomass content was constant in both chronic and acute toxicity experiments, but the ammonia oxidizing activity, measured as ammonia uptake rate, was different. The AOB community structural changes due to the copper presence were evaluated by multivariate analysis of the DGGE bands profiles. The chronic contamination caused a change in the AOB community compared to the control. In contrast, acute inputs led to a temporary change in the AOB community, after which the community was similar to the control. Recovery after acute intoxication was achieved after 72 h. The present study reports on the effects of chronic and acute copper contamination on the ammonia uptake ability of the AO microorganisms and the structure of the AOB community in a wastewater system and, as a consequence, gives indications on the response of wastewater plants under similar conditions.
References
91/271/EC, Concerning urban waste treatment. 1991, Council Directive
2000/60/EC, Establishing a framework for Community action in the field of water policy, EuropeanParliamen and Council, 2000
Avrahami S, Conrad R (2003) Patterns of community change among ammonia oxidizers in meadow soils upon long-term incubation at different temperatures. Appl Environ Microb 69(10):6152–6164
Bothe H, Jost G, Schloter M, Ward BB, Witzel K-P (2000) Molecular analysis of ammonia oxidation and denitrification in natural environments. FEMS Microbiol Rev 24(5):673–690
Broos K, Uyttebroek M, Mertens J, Smolders E (2004) A survey of symbiotic nitrogen fixation by white clover grown on metal contaminated soils. Soil Biol Biochem 36(4):633–640
Bundy JG, Paton GI, Campbell CD (2004) Combined microbial community level and single species biosensor responses to monitor recovery of oil polluted soil. Soil Biol Biochem 36(7):1149–1159
Cabrera G, Pérez R, Gómez JM, Ábalos A, Cantero D (2006) Toxic effects of dissolved heavy metals on Desulfovibrio vulgaris and Desulfovibrio sp. strains. J Hazard Mater 135(1–3):40–46
Chang LW (1996) In: Chang LW, Magos L, Suzuki T (eds) Toxicology of Metals, vol. I. CRC Lewis, New York
Chen F, Dustman WA, Hodson RE (1999) Microscopic detection of the toluene dioxygenase gene and its expression inside bacterial cells in seawater using prokaryotic in situ PCR. Hydrobiologia 401:131–138
Demanou J, Sharma S, Weber A, Wilke B-M, Njine T, Monkiedje A, Munch JC, Schloter M (2006) Shifts in microbial community functions and nitrifying communities as a result of combined application of copper and mefenoxam. FEMS Microbiol Lett 260(1):55–62
Dumestre A, Sauvé S, McBride M, Baveye P, Berthelin J (1999) Copper speciation and microbial activity in long-term contaminated soils. Arch Environ Contam Toxicol 36:124–131
Ehrlich HL (1997) Microbes and metals. Appl Microbiol Biotechnol 48(6):687–692
Giller KE, Witter E, McGrath SP (1998) Toxicity of heavy metals to microorganisms and microbial processes in agricultural soils: a review. Soil Biol Biochem 30(10–11):1389–1414
Harms G, Layton A, Dionisi H, Gregory I, Garrett V, Hawkins S, Robinson K, Sayler G (2003) Real-time PCR quantification of nitrifying bacteria in a municipal wastewater treatment plant. Environ Sci Technol 37(2):343–351
Hoshino T, Noda N, Tsuneda S, Hirata A, Inamori Y (2001) Direct detection by in situ PCR of the amoA gene in biofilm resulting from a nitrogen removal process. Appl Environ Microb 67(11):5261–5266
Hoshino T, Tsuneda S, Hirata A, Inamori Y (2003) In situ PCR for visualizing distribution of a functional gene “amoA” in a biofilm regardless of activity. J Biotechnol 105(1–2):33–40
Hu Z, Chandran K, Grasso D, Smets BF (2004) Comparison of nitrification inhibition by metals in batch and continuous flow reactors. Water Res 38(18):3949–3959
Lawlor K, Knight BP, Barbosa-Jefferson VL, Lane PW, Lilley AK, Paton GI, McGrath SP, O’Flaherty SM, Hirsch PR (2000) Comparison of methods to investigate microbial populations in soils under different agricultural management. FEMS Microbiol Ecol 33(2):129–137
Layton AC, Dionisi H, Kuo H-W, Robinson KG, Garrett VM, Meyers A, Sayler GS (2005) Emergence and competitive dominant ammonia-oxidizer bacterial populations in a full-scale industrial. Appl Environ Microb 71(2):1105–1108
Li B, Bishop P (2003) Structure–function dynamics and modeling analysis of the micro-environment of activated sludge floc. Water Sci Technol 47(11):267–273
Li B, Bishop PL (2004) Micro-profiles of activated sludge floc determined using microelectrodes. Water Res 38(5):1248–1258
Li Z, Xu J, Tang C, Wu J, Muhammad A, Wang H (2006) Application of 16S rDNA-PCR amplification and DGGE fingerprinting for detection of shift in microbial community diversity in Cu-, Zn-, and Cd-contaminated paddy soils. Chemosphere 62(8):1374–1380
Manz W, Amann R, Ludwig W, Wagner M, Schleifer K-H (1992) Phylogenetic oligodeoxynucleotide probes for the major subclasses of Proteobacteria: problems and solutions. Syst Appl Microbiol 15:593–600
Nicolaisen MH, Ramsing NB (2002) Denaturing gradient gel electrophoresis (DGGE) approaches to study the diversity of ammonia-oxidizing bacteria. J Microbiol Methods 50(2):189–203
Nies D (1999) Microbial heavy-metal resistance. Appl Microbiol Biotechnol 51(6):730–750
O’Mullan GD, Ward BB (2005) Relationship of temporal and spatial variabilities of ammonia-oxidizing bacteria to nitrification rates in Monterey Bay, California. Appl Environ Microbiol 71(2):697–705
Principi P, Villa F, Bernasconi M, Zanardini E (2006) Metal toxicity in municipal wastewater activated sludge investigated by multivariate analysis and in situ hybridization. Water Res 40(1):99–106
Purkhold U, Pommerening-Röser A, Juretschko S, Schmid MC, Koops H-P, Wagner M (2000) Phylogeny of all recognized species of ammonia oxidizers based on comparative 16S rRNA and amoA sequence analysis: implications for molecular diversity surveys. Appl Environ Microbiol 66(12):5368–5382
Rajapaksha RMCP, Tobor-Kapłon MA, Bååth E (2004) Metal toxicity affects fungal and bacterial activities in soil differently. Appl Environ Microbiol 70:5
Ranalli G, Alfano G, Belli C, Lustrato G, Colombini MP, Bonaduce I, Zanardini E, Abbruscato P, Cappitelli F, Sorlini C (2005) Biotechnology applied to cultural heritage: biorestoration of frescoes using viable bacterial cells and enzymes. J Appl Microbiol 98(1):73–83
Ranjard L, Echairi A, Nowak V, Lejon DPH, Nouaim R, Chaussod R (2006) Field and microcosm experiments to evaluate the effects of agricultural Cu treatment on the density and genetic structure of microbial communities in two different soils. FEMS Microbiol Ecol 58(2):303–315
Rasband W (2008) ImageJ 1997–2007. U.S. National Institutes of Health, Bethesda, Maryland, USA
Rotthauwe JH, Witzel KP, Liesack W (1997) The ammonia monooxygenase structural gene amoA as a functional marker: molecular fine-scale analysis of natural ammonia-oxidizing populations. Appl Environ Microbiol 63(12):4704–4712
Sauvé S, Dumestre A, McBride M, Gillett JW, Berthelin J, Hendershot W (1999) Nitrification potential in field-collected soils contaminated with Pb or Cu. Appl Soil Ecol 12(1):29–39
Smit E, Leeflang P, Wernars K (1997) Detection of shifts in microbial community structure and diversity in soil caused by copper contamination using amplified ribosomal DNA restriction analysis. FEMS Microbiol Ecol 23(3):249–261
Tobor-Kapłon M, Bloem J, de Ruiter P (2006) Functional stability of microbial communities from long-term stressed soils to additional disturbance. Environ Toxicol Chem 25(8):1993–1939
Turpeinen R, Kairesalo T, Häggblom MM (2004) Microbial community structure and activity in arsenic-, chromium- and copper-contaminated soils. FEMS Microbiol Ecol 47(1):39–50
Wang Y, Shi J, Wang H, Lin Q, Chen X, Chen Y (2007) The influence of soil heavy metals pollution on soil microbial biomass, enzyme activity, and community composition near a copper smelter. Ecotoxicol Environ Saf 67(1):75–81
Zhang T, Fang HHP (2000) Digitization of DGGE (denaturing gradient gel electrophoresis) profile and cluster analysis of microbial communities. Biotechnol Lett 22:399–405
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Principi, P., Villa, F., Giussani, B. et al. The Effect of Copper on The Structure of the Ammonia-Oxidizing Microbial Community in an Activated Sludge Wastewater Treatment Plant. Microb Ecol 57, 215–220 (2009). https://doi.org/10.1007/s00248-008-9432-5
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DOI: https://doi.org/10.1007/s00248-008-9432-5