Journal of Soils and Sediments

, Volume 10, Issue 3, pp 537–544 | Cite as

Effect of sulfonamide antibiotics on microbial diversity and activity in a Californian Mollic Haploxeralf

  • Iris R. Gutiérrez
  • Naoko Watanabe
  • Thomas Harter
  • Bruno Glaser
  • Michael Radke



Up to 90% of antibiotics that are fed to livestock are excreted unaltered or as metabolites and thus are present in manure. By application of manure as fertilizer, veterinary antibiotics can reach soil and groundwater. The aim of this study is to determine the effect of three commonly used (and simultaneously applied) sulfonamide antibiotics on both function and structural diversity of soil microorganisms. To this end, the activity of the enzymes urease and dehydrogenase was determined, and the composition of phospholipid fatty acids (PLFA) was analyzed.

Materials and methods

Soil and manure were sampled at a dairy farm located in the Northern San Joaquin Valley, California, USA. Soil (700 g) was amended with either mineral water only (W-treatments), liquid manure (M-treatments), or with glucose solution (G-treatments). Each of these soil treatments was mixed with a cocktail of three sulfonamides: sulfadimethoxine (SDT), sulfamethoxazole (SMX), and sulfamethazine (SMZ) at five total concentration levels ranging from 0 (control) to 900 µg g dm −1 . After 24, 48, 96, 168, 264, 384, and 504 h, UA and DHA were determined; PLFA composition in selected samples was analyzed at t = 168 h and 504 h of incubation.

Results and discussion

In the G-treatments, urease activity decreased with higher sulfonamide concentrations; no effect was observed when no glucose was added (W-treatments). While a dose–response relationship was observed for urease activity after 168 h, a similar inhibition was measured after 380 h at all sulfonamide concentrations. Sulfonamides also reduced dehydrogenase activity in the G-treatments, but results are less conclusive than for urease. With increasing sulfonamide concentration, microbial and bacterial biomass decreased in the G-treatments compared to the control at 168 h. Sulfonamides caused a relative community shift towards gram-negative bacteria and towards an increased proportion of fungal biomass. Strong inhibition of urease by manure (M-treatments) was observed even without the addition of sulfonamides.


Sulfonamides clearly affected both the function and structural diversity of the soil microbial community over at least 16 days. The soil microbial community was affected by sulfonamides even at a relatively low concentration, although this soil receives regular input of manure that contains several antibiotics. Further research is needed addressing both long-term effects and lower sulfonamide concentrations under dynamic boundary conditions.


Antibiotics Dehydrogenase Enzyme activity Phospholipid fatty acids Soil microorganisms Urease 



This study was financially supported by grants from the Bavaria California Technology Center (BaCaTeC) to MR and the German Academic Exchange Service (DAAD) to IRG. We thank two anonymous reviewers for their constructive comments on earlier versions of this manuscript.

Supplementary material

11368_2009_168_MOESM1_ESM.doc (84 kb)
ESM 1 Electronic supplementary material. (DOC 84 kb)


  1. Böhme L, Langer U, Böhme F (2005) Microbial biomass, enzyme activities and microbial community structure in two European long-term field experiments. Agric Ecosyst Environ 109:141–152CrossRefGoogle Scholar
  2. Bol R, Kandeler E, Amelung W, Glaser B, Marx MC, Preedy N, Lorenz K (2003) Short-term effects of dairy slurry amendment on carbon sequestration and enzyme activities in a temperate grassland. Soil Biol Biochem 35:1411–1421CrossRefGoogle Scholar
  3. Esiobu N, Armenta L, Ike J (2002) Antibiotic resistance in soil and water environments. Int J Environ Health Res 12:133–144CrossRefGoogle Scholar
  4. Frostegard A, Tunlid A, Baath E (1991) Microbial biomass measured as total lipid phosphate in soils of different organic content. J Microbiol Meth 14:151–163CrossRefGoogle Scholar
  5. Hackl E, Pfeffer M, Donat C, Bachmann G, Zechmeister-Boltenstern S (2005) Composition of the microbial communities in the mineral soil under different types of natural forest. Soil Biol Biochem 37:661–671CrossRefGoogle Scholar
  6. Halling-Sørensen B, Nors Nielsen S, Lanzky PF, Ingerslev F, Holten Lützhøft HC, Jørgensen SE (1998) Occurrence, fate and effects of pharmaceutical substances in the environment—a review. Chemosphere 36:357–393CrossRefGoogle Scholar
  7. Hammesfahr U, Heuer H, Manzke B, Smalla K, Thiele-Bruhn S (2008) Impact of the antibiotic sulfadiazine and pig manure on the microbial community structure in agricultural soils. Soil Biol Biochem 40:1583–1591CrossRefGoogle Scholar
  8. Hamscher G, Pawelzick HT, Höper H, Nau H (2005) Different behaviour of tetracyclines and sulfonamides in sandy soils after repeated fertilization with liquid manure. Environ Toxicol Chem 24:861–868CrossRefGoogle Scholar
  9. Harter T, Davis H, Mathews MC, Meyer RD (2002) Shallow groundwater quality on dairy farms with irrigated forage crops. J Contam Hydrol 55:287–315CrossRefGoogle Scholar
  10. Heise J, Höltge S, Schrader S, Kreuzig R (2006) Chemical and biological characterization of non-extractable sulfonamide residues in soil. Chemosphere 65:2352–2357CrossRefGoogle Scholar
  11. Kahle M, Stamm C (2007) Time and pH-dependent sorption of the veterinary antimicrobial sulfathiazole to clay minerals and ferrihydrite. Chemosphere 68:1224–1231CrossRefGoogle Scholar
  12. Kandeler E, Gerber H (1988) Short-term assay of soil urease activity using colorimetric determination of ammonium. Biol Fertil Soils 6:68–72CrossRefGoogle Scholar
  13. Kandeler E, Kampichler C, Horak O (1996) Influence of heavy metals on the functional diversity of soil microbial communities. Biol Fertil Soils 23:299–306CrossRefGoogle Scholar
  14. Kandeler E, Stemmer M, Klimanek EM (1999) Response of soil microbial biomass, urease and xylanase within particle size fractions to long-term soil management. Soil Biol Biochem 31:261–273CrossRefGoogle Scholar
  15. Klose S, Tabatabai MA (1999) Urease activity of microbial biomass in soils. Soil Biol Biochem 31:205–211CrossRefGoogle Scholar
  16. Kong WD, Zhu YG, Fu BJ, Marschner P, He JZ (2006) The veterinary antibiotic oxytetracycline and cu influence functional diversity of the soil microbial community. Environ Pollut 143:129–137CrossRefGoogle Scholar
  17. Kotzerke A, Sharma S, Schauss K, Heuer H, Thiele-Bruhn S, Smalla K, Wilke BM, Schloter M (2008) Alterations in soil microbial activity and N-transformation processes due to sulfadiazine loads in pig-manure. Environ Pollut 153:315–322CrossRefGoogle Scholar
  18. Madigan MT, Martinko JM, Dunlap PV, Clark DP (2009) Brock biology of microorganisms. 12th international ed., Pearson, San FranciscoGoogle Scholar
  19. Sarmah AK, Meyer MT, Boxall ABA (2006) A global perspective on the use, sales, exposure pathways, occurrence, fate and effects of veterinary antibiotics (VAs) in the environment. Chemosphere 65:725–759CrossRefGoogle Scholar
  20. Scheffer P, Schachtschabel P, Blume H-P, Brümmer G, Hartge KH, Schwertmann U, Auerswald K, Beyer L, Fischer WR, Kögel-Knaber I, Renger M, Strebel O (1998) Lehrbuch der Bodenkunde, 14th edn. Ferdinand Enke, Stuttgart, p 494Google Scholar
  21. Schmitt H, Haapakangas H, van Beelen P (2005) Effects of antibiotics on soil microorganisms: time and nutrients influence pollution-induced community tolerance. Soil Biol Biochem 37:1882–1892CrossRefGoogle Scholar
  22. Schmitt H, van Beelen P, Tolls J, van Leeuwen CL (2004) Pollution-induced community tolerance of soil microbial communities caused by the antibiotic sulfachloropyridazine. Environ Sci Technol 38:1148–1153CrossRefGoogle Scholar
  23. Schmitt A, Glaser B, Borken W, Matzner E (2008) Repeated freeze–thaw cycles changed organic matter quality in a temperate forest soil. J Plant Nutr Soil Sci 171:707–718CrossRefGoogle Scholar
  24. Snedecor GW, Cochran WT (1989) Statistical methods. Ames, IowaGoogle Scholar
  25. Thalmann A (1968) Zur Methodik der Bestimmung der Dehydrogenaseaktivität im Boden mittels Triphenyltetrazoliumchlorid (TTC). Landwirtsch Forsch 21:249–258Google Scholar
  26. Thiele-Bruhn S, Beck IC (2005) Effects of sulfonamide and tetracycline antibiotics on soil microbial activity and microbial biomass. Chemosphere 59:457–465CrossRefGoogle Scholar
  27. Watanabe N, Harter T, Bergamaschi BA (2010) Environmental occurrence of antibiotics from dairy farms. In pressGoogle Scholar
  28. Watanabe N, Harter TH, Bergamaschi BA (2008) Environmental occurrence and shallow ground water detection of the antibiotic monensin from dairy farms. J Environ Qual 37:78–85CrossRefGoogle Scholar
  29. Zelles L (1999) Fatty acid patterns of phospholipids and lipopolysaccharides in the characterisation of microbial communities in soil: a review. Biol Fertil Soils 29:111–129CrossRefGoogle Scholar
  30. Zielezny Y, Groeneweg J, Vereecken H, Tappe W (2006) Impact of sulfadiazine and chlorotetracycline on soil bacterial community structure and respiratory activity. Soil Biol Biochem 38:2372–2380CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Iris R. Gutiérrez
    • 1
  • Naoko Watanabe
    • 2
  • Thomas Harter
    • 2
  • Bruno Glaser
    • 3
  • Michael Radke
    • 1
  1. 1.Department of Hydrology, BayCEERUniversity of BayreuthBayreuthGermany
  2. 2.Department of Land, Air and Water ResourcesUniversity of CaliforniaDavisUSA
  3. 3.Department of Soil Physics, BayCEERUniversity of BayreuthBayreuthGermany

Personalised recommendations