Abstract
We assessed potential toxicity of fungicides to natural bacterial communities from a constructed wetland, located in southern Sweden, and compared the sensitivity of two endpoints indicating bacterial activity, leucine incorporation, and potential denitrification, in detecting toxicity. The effects of eight fungicides (benomyl, carbendazim, carboxin, captan, cycloheximide, fenpropimorph, propiconazole, and thiram), two bactericides (bronopol and chlortetracycline) as controls, and one reference compound (3,5-dichlorophenol), were tested in a water-sediment microcosm set-up. Leucine incorporation was measured in both the water and sediment column, while potential denitrification was measured for the entire microcosm. The bactericides and the reference compound gave sigmoid concentration–response curves for both endpoints in all but one case. The fungicides thiram, captan, and benomyl, and to a lesser extent fenpropimorph and propiconazole had quantifiable toxic effects on leucine incorporation, with EC50 values ranging from 3 to 70 mg l−1, while carbendazim, carboxin, and cycloheximide had little effect at the investigated concentrations. Only thiram and captan inhibited potential denitrification; the other fungicides showed no quantifiable effect. A greater toxic effect on leucine incorporation was recorded for bacterial communities associated with the water column, compared to the sediment column, for all tested compounds. Leucine incorporation was the more sensitive method for toxicity assessment of bacterial communities, and also allowed for a rapid and simple way of comparing exposure in the sediment and water column, making it an attractive standard method for community based toxicological assays in aquatic environments.
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References
Aldén Demoling L, Bååth E (2008) The use of leucine incorporation to determine the toxicity of phenols to bacterial communities extracted from soil. Appl Soil Ecol 38:34–41
Aldén Demoling L, Bååth E, Greve G, Wouterse M, Schmitt H (2009) Effects of sulfamethoxazole on soil microbial communities after adding substrates. Soil Biol Biochem 41:840–848
Bååth E, Petterson M, Söderberg KH (2001) Adaption of a rapid and economical microcentrifugation method to measure thymidine and leucine incorporation by soil bacteria. Soil Biol Biochem 33:1571–1574
Bastviken D, Tranvik L (2001) The leucine incorporation method estimates bacterial growth equally well in both oxic and anoxic lake water. Appl Environ Microbiol 67:2916–2921
Brandt KK, Jørgensen NOG, Nielsen TH, Winding A (2004) Microbial community-level toxicity testing of linear alkylbenzene sulfonates in aquatic microcosms. FEMS Microbiol Ecol 49:229–241
Brandt KK, Sjøholm OR, Krogh KA, Halling-Sørensen B, Nybroe O (2009) Increased pollution-induced bacterial community tolerance to sulfadiazine in soil hotspots amended with artificial root exudates. Environ Sci Technol 43:2963–2968
Broos K, Mertens J, Smolders E (2005) Toxicity of heavy metals in soil assessed with various soil microbial and plant growth assays: as comparative study. Environ Toxicol Chem 24:634–640
Cameron HJ, Julian GR (1984) The effects of four commonly used fungicides on the growth of Cyanobacteria. Plant Soil 78:409–415
Carlson JC, Mabury SA (2006) Dissipation kinetics and mobility of chlortetracycline, tylosin, and monesin in an agricultural soil in Northumberland County, Ontario, Canada. Environ Toxicol Chem 25:1–10
De Zwart D, Posthuma L (2005) Complex mixture for single and multiple species: proposed methodologies. Environ Toxicol Chem 24:2665–2676
Díaz-Raviña M, Bååth E (2001) Response of soil bacterial communities pre-exposed to different metals and reinoculated in an unpolluted soil. Soil Biol Biochem 33:241–248
Domsch KH, Jagnow G, Anderson T-H (1983) An ecological concept for assessment of side-effects of agrochemicals on soil-microorganisms. Residue Rev 86:65–105
Howard PH, Meylan WM (1997) Handbook of physical properties of organic chemicals. CRC Press, Boca Raton
ISO 15522, International Organization of Standardization (1999) Water quality-determination of the inhibitory effect of water constituents on the growth of activated sludge microorganisms. Geneva, Switzerland
Johnsen K, Jacobsen CS, Torsvik V, Sørensen J (2001) Pesticide effects on bacterial diversity in agricultural soils—a review. Biol Fertil Soils 33:443–453
Kah M, Beulke S, Brown CD (2007) Factors influencing degradation of pesticides in soil. J Agric Food Chem 55:4487–4492
Kahru A, Tomson K, Pall T, Külm I (1996) Study of toxicity of pesticides using luminiscent bacteria Photobacterium phosphoreum. Water Sci Technol 33:147–154
Kinney CA, Mandernack KW, Moiser AR (2005) Laboratory investigations into the effects of the pesticides mancozeb, chlorothalonil, and prosulfuron on nitrous oxide and nitric oxide production in fertilized soil. Soil Biol Biochem 37:837–850
Kreuger J, Peterson M, Lundgren E (1999) Agricultural inputs of pesticide residues to stream and pond sediments in a small catchment in Southern Sweden. Bull Environ Contam Toxicol 62:55–62
Larson F, Petersen DG, Dahllöf I, Sundbäck K (2007) Combined effects of an antifouling biocide and nutrient status on a shallow-water microbenthic community. Aquat Microb Ecol 48:277–294
Loffhagen N, Härtig W, Babel W (2003) Energization of Comamonas testosteroni ATCC 17454 for indicating toxic effects of chlorophenoxy herbicides. Arch Environ Contam Toxicol 45:317–323
Lovley D (2003) Cleaning up with genomics: applying molecular biology to bioremediation. Nature Rev 1:35–44
Mackay D, Shiu WY, Ma K-C, Lee SC (2006) Handbook of physical–chemical properties and environmental fate for organic chemicals, vol I–IV, 2nd edn. CRC Press, New York
Mankiewicz-Boczek J, Nałęcz-Jawecki G, Drobniewska A, Kaza M, Sumorok B, Izydorczyk K, Zalewski M, Sawicki J (2008) Application of a microbiotests battery for complete toxicity assessment of rivers. Ecotoxicol Environ Saf 71:830–836
Maraldo K, Dahllöf I (2004) Indirect estimation of degradation time for zinc pyrithione and copper pyrithione in seawater. Mar Poll Bull 48:894–901
Masunaga S, Susarla S, Gundersen JL, Yonezawa Y (1996) Pathway and rate of chlorophenol transformation in anaerobic estuarine sediment. Environ Sci Technol 30:1253–1260
Miller MN, Zebarth BJ, Dandie CE, Burton DL, Goyer C, Trevors JT (2008) Crop residue influence on denitrification, N2O emissions and denitrifier community abundance in soil. Soil Biol Biochem 40:2553–2562
National Library of Medicine (2003) US ChemiDplus Lite. http://chem.sis.nlm.nih.gov/chemidplus/chemidlite.jsp. Accessed 21 Feb 2009
Park S, Choi K (2008) Hazard assessment of commonly used agricultural antibiotics on aquatic ecosystems. Ecotoxicology 17:526–538
Pell M, Stenberg B, Torstensson L (1998) Potential denitrification and nitrification tests for evaluation of pesticide effects in soil. Ambio 27:24–28
Petersen DG, Dahllöf I, Nielsen LP (2004) Effects of zinc pyrithione and copper pyrithione on microbial community function and structure in sediments. Environ Toxicol Chem 23:921–928
Pietikäinen J, Pettersson M, Bååth E (2005) Comparison of temperature effects on soil respiration and bacterial and fungal growth rates. FEMS Microbiol Ecol 52:49–58
Rousk J, Aldén Demoling L, Bååth E (2009) Contrasting short-term antibiotic effects on respiration and bacterial growth compromises the validity of the selective respiratory inhibition technique to distinguish fungi and bacteria. Microb Ecol 58:75–85
Schmidt IK, Ruess L, Bååth E, Michelsen A, Ekelund F, Jonasson S (2000) Long-term manipulation of the microbes and microfauna of two subarctic heaths by addition of fungicide, bactericide, carbon and fertilizer. Soil Biol Biochem 32:707–720
Seitzinger S, Harrison JA, Böhlke JK, Bouwman AF, Lowrance R, Peterson B et al (2006) Denitrification across landscapes and waterscapes: a synthesis. Ecol Appl 16:2064–2090
Smith DC, Azam F (1992) A simple, economical method for measuring bacterial protein synthesis rates in seawater using tritiated-leucine. Mar Microb Food Webs 6:107–114
Sundbäck K, Petersen GD, Dahllöf I, Larson F (2007) Combined nutrient-toxicant effects on a shallow-water marine sediment system: sensitivity and resilience of ecosystem functions. Mar Ecol Prog Ser 330:13–30
Thiere G, Milenkovski S, Lindgren P-E, Sahlén G, Berglund O, Weisner SEB (2009) Wetland creation in agricultural landscapes: biodiversity benefits on local and regional scales. Biol Conserv 142:965–973
Tomlin C (1997) The pesticide manual. British Crop Protection Council, Surrey
Wang HF, Provan GJ, Helliwell K (2002) Determination of bronopol and its degradation product by HPLC. J Pharmaceut Biomed Anal 29:387–392
Warren N, Allan IJ, Carter JE, House WA, Parker A (2003) Pesticides and other micro-organic contaminants in freshwater sedimentary environments—a review. Appl Geochem 18:159–194
Welp G, Brümmer G (1997) Toxicity of increased amounts of chemicals and the dose–response curves for heterogeneous microbial populations in soil. Ecotoxicol Environ Saf 37:37–44
Welp G, Brümmer G (1999) Effects of organic pollutants on soil microbial activity: the influence of sorption, solubility and speciation. Ecotoxicol Environ Saf 43:83–90
Widenfalk A, Svensson MJ, Goedkoop W (2004) Effects of the pesticides captan, delthamethrin, isoproturon, and pirimicarb on the microbial community of a freshwater sediment. Environ Toxicol Chem 23:1920–1927
Widenfalk A, Bertilsson S, Sundh I, Goedkoop W (2008) Effects of pesticides on community composition and activity of sediment microbes—responses at various levels of microbial community organization. Environ Poll 152:576–584
Worthing CR, Walker SB (1983) The pesticide manual. British Crop Protection Council, Surrey
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This study was partly funded by FORMAS, the Swedish Research Council for Environment, Agricultural Sciences and Spatial Planning (grant nos. 25/2-2001-2677 and 214-2004-1678), and the Crafoord Foundation (grant no. 20050957). The authors would like to thank Johanna Stadmark and Lars Leonardsson for technical support.
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Milenkovski, S., Bååth, E., Lindgren, PE. et al. Toxicity of fungicides to natural bacterial communities in wetland water and sediment measured using leucine incorporation and potential denitrification. Ecotoxicology 19, 285–294 (2010). https://doi.org/10.1007/s10646-009-0411-5
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DOI: https://doi.org/10.1007/s10646-009-0411-5