Research Article

Environmental Science and Pollution Research

, Volume 20, Issue 5, pp 3435-3446

First online:

Open Access This content is freely available online to anyone, anywhere at any time.

Application of a battery of biotests for the determination of leachate toxicity to bacteria and invertebrates from sewage sludge-amended soil

  • Anna MalaraAffiliated withFaculty of Chemistry, Maria Curie-Skłodowska University
  • , Patryk OleszczukAffiliated withFaculty of Chemistry, Maria Curie-Skłodowska University Email author 


The objective of the study was to determine the leachates toxicity from sewage sludge-amended soils (sandy and loamy). Samples originated from a plot experiment realized over a period of 29 months. Two types of soil were fertilized with sewage sludges at the dose of 3 % (90 t/ha). Soil samples were taken after 0, 7, 17, and 29 months from the application of sewage sludges. Leachates were obtained according to the EN 12457–2 protocol. The following commercial tests were applied for the estimation of the toxicity: Microtox (Vibrio fischeri), Microbial assay for toxic risk assessment (ten bacteria and one yeast), Protoxkit F™ (Tetrahymena thermophila), Rotoxkit F™ (Brachionus calyciflorus), and Daphtoxkit F™ (Daphnia magna). The test organisms displayed varied toxicity with relation to the soils amended with sewage sludges. The toxicity of the leachates depended both on the soil type and on the kind of sewage sludge applied. Notable differences were also observed in the sensitivity of the test organisms to the presence of sewage sludge in the soil. The highest sensitivity was a characteristic of B. calyciflorus, while the lowest sensitivity to the presence of the sludges was revealed by the protozoa T. thermophila. Throughout the periods of the study, constant variations of toxicity were observed for most of the test organisms. The intensity as well as the range of those variations depended both on the kind of test organism and on the kind of sludge and soil type. In most cases, an increase of the toxicity of soils amended with the sewage sludges was observed after 29 months of the experiment.


Sewage sludges Toxicity Bioassays Biotests Ecotoxicology