Toxicity of pesticides on soil microbes: new risk assessment procedures for standardized risk assessment

Toxicity of pesticides onto non target organisms is an indispensable part of the pesticide regulatory framework. Although advanced protocols and tools have been developed and used for the assessment of the toxicity of pesticides on aquatic organisms and terrestrial macro-organisms this is not the case for soil microorganisms. Despite their pivotal role in soil ecosystem functioning, the assessment of the soil microbial toxicity of pesticides is still done using two out-of-date microbial tests, C and N transformation (OECD 217 and 216 respectively).  Methodological advances have drastically changed the scene in soil microbiology.  The implementation of ISO standardized tools in the environmental risk assessment of pesticides will enable the comprehensive assessment of the toxicity of pesticides on soil microbes. Our group has lead research work for the revision of the current regulatory framework and proposed new procedures for assessing the toxicity of pesticides on soil microorganisms. Arbuscular mycorrhizal (AM) fungi and ammonia-oxidizing microorganisms are used as toxicity endpoints in a tiered assessment approach spanning from in-vitro testing (Tier I) to field experimentation (Tier III). Our current focus is:

(i) on the development of in vitro tests in order to accurately assess the toxicity of pesticides on ammonia oxidizing microorganisms and AM fungi

(ii) on the study of the impact of pesticides on soil microorganisms looking at the food-web level (predators - prey system).


  1. ERA-NETplusDevelopment and implementation of innovative tools to estimate the ecotoxicological impact of low dose pesticide application in agriculture on soil functional microbial diversity – ECOFUN-MICROBIODIV. Funding: EU/DLR, Duration: 1/11/2010-30/9/12, Website:
  2. IAPP Marie Curie projectPesticides – Felicity or curse for the soil microbes (Acronym LOVE-TO-HATE). Funding: EC-FP7, Duration: 1/1/2013 – 31/12/2016, Website
  3. CARIPLO project, Synthetic and Natural Agrochemical compounds: ecological impacts on the soil ecosystem and effects on plant production. Funding Body: The CARIPLO Foundation,. Duration: 1/2012 – 12/2014, Coordinator: Universita Cattolica del Sacro Cuore, Sede di Piacenza
  4. IUPAC project. Advances on the Assessment of Pesticides' Soil Microbial toxicity: New research and regulatory aspects in light of the recent methodological advances. Funding: IUPAC Division of the Environment, Duration: 2015 - 2018, Website:












  1. Spyrou I.M., Karpouzas D.G., Menkissoglu-Spiroudi, U., (2009). Do botanical pesticides alter the structure of the soil microbial community. Microbial Ecology 58: 715-727
  2. Karpouzas D.G., Rousidou C., Papadopoulou K.K, Bekris F., Zervakis G., Singh B.K., Ehaliotis C., (2009). Effect of continuous olive mill wastewater applications, in the presence and absence of N fertilization, on the structure of rhizopshere soil fungal communities. FEMS Microbiology Ecology 70: 56-69
  3. Karpouzas, D.G., Singh B.K., (2009) Chapter 5: Application of fingerprinting molecular methods in bioremediation studies. In BioremediationMethods in Molecular Biology 599, Cummings S.P. Ed., Humana Press Inc., pp 69-88.
  4. Rousidou C., Papadopoulou KK., Zervakis G., Singh B.K., Ehaliotis C., Karpouzas, D.G., (2010) Fractioned land application of diluted olive mill wastewater induces changes in the structure of the soil microbial community. European Journal of Soil Biology 46: 34-4
  5. Karpouzas D.G., Ntougias S., Iskidou E., Rousidou C., Papadopoulou K.K., Zervakis G., Ehaliotis C., (2010) The effects of soil application of olive mill wastewaters on functional soil bacterial communities. Applied Soil Ecology 45: 101-111.
  6. Omirou, C. Rousidou, F. Bekris, K.K. Papadopoulou, C. Ehaliotis, U. Menkissoglu-Spiroudi, D.G. Karpouzas. (2011) Τhe impact of biofumigation and chemical fumigation methods on the structure and function of the soil microbial community. Microbial Ecology 61: 201-213
  7. Papadopoulou E.S., Karpouzas D.G., Menkissoglu-SpiroudiU., (2011) Extraction parameters significantly influence and the quantity and the profile of PLFAs extracted from soil. Microbial Ecology 62: 704-714
  8. Karpouzas D.G., Karatasas A., Spyridaki E., Rousidou C., Bekris F., Ehaliotis C., Papadopoulou K.K. (2011) Impact of a beneficial and of a pathogenic Fusarium strain on the fingerprinting-based structure of microbial communities in tomato (Lycopersicon esculentum) rhizosphere. European Journal of Soil Biology 47(6): 400-408
  9. Ipsilantis I., Samourelis C., Karpouzas D.G., (2012) The impact of biological pesticides on arbuscular mycorrhizal fungi. Soil Biology and Biochemistry 45: 147-155
  10. Puglisi E., Vasileiadis S., Demiris, C., Bassi D., Karpouzas D.G., Capri E., Cocconcelli PS., Trevisan M., (2012) Fungicides impact on the diversity and function of non-target ammonia oxidizing microorganisms residing in a litter soil cover. Microbial Ecology 64: 692-701
  11. Martin-Laurent F, Kandeler E., Pertic I, Djuric S, Karpouzas D.G., (2013) ECOFUN-MICROBIODIV: an FP7 European project for developing and evaluating innovative tools for assessing the impact of pesticides on soil functional microbial diversity-towards new pesticide registration regulation? Environmental Science and Pollution Research 20:1203-1205
  12. Rousidou C., Papadopoulou E., Kortsinidou M., Giannakou I.O., Singh B.K., Menkissoglu-Spiroudi, U., and Karpouzas D.G.,(2013) Bio-pesticides: Harmful or harmless to ammonia oxidizing microorganisms? The case of a Paecilomyces lilacinus-based nematicide. Soil Biology & Biochemistry 67:98-105
  13. Karpouzas D.G., Papadopoulou, E, Ipsilantis I., Petric I., Udikovic-Kolic N., Djuric S., Kandeler E., Menkissoglu-Spiroudi U., Martin-Laurent F., (2013) Effects of nicosulfuron on the abundance and diversity of arbuscular mycorrhizal fungi used as indicators of pesticide soil microbial toxicity. Ecological Indicators 39: 44-53
  14. Karpouzas, D.G., Kandeler, E., Bru, D., Friedel, I., Auer, Y., Kramer, S., Vasileiadis, S., Petric, I., Udikovic-Kolic, N., Djuric, S., Martin-Laurent, F. (2014) A tiered assessment approach based on standardized methods to estimate the impact of nicosulfuron on the abundance and function of the soil microbial community. Soil Biology and Biochemistry 75: 282-291
  15. Petric, I., Karpouzas, D.G., Bru, D., Udikovic-Kolic, N., Kandeler, E., Djuric, S.,Martin-Laurent, F (2016) Nicosulfuron application in agricultural soils drives the selection towards NS-tolerant microorganisms harboring various levels of sensitivity to nicosulfuron. Environmental Science and Pollution Research 23(5): 4320-4333
  16. Papadopoulou E.S., Tsachidou P., Sulowic S., Menkissoglu-Spiroudi U., Karpouzas D.G., (2016) Land spreading of wastewaters from the fruit packaging industry and potential effects on soil microbes: Effects of the antioxidant ethoxyquin and its metabolites on ammonia oxidizers. Applied and Environmental Microbiology 82: 747-755
  17. Papazlatani C., Rousidou C., Katsoula A., Kolyvas M., Genitsaris S., Papadopoulou K.K.., Karpouzas D.G., (2016) Assessment of the impact of the fumigant dimethyl disulfide on the dynamics of major fungal plant pathogens in greenhouse soils. European Journal of Plant Pathology 146 (2): 391-400
  18. Karpouzas D.G., Tsiamis G., Trevisan M., Ferrari F., Malandain C., Sibourg O. Martin-Laurent F., (2016) 'LOVE TO HATE'-Pesticides: Felicity or curse for the soil microbial community? An FP7 IAPP Marie Curie project aiming to establish tools for the assessment of the mechanisms controlling the interactions of pesticides with soil microorganisms. Environmental Science and Pollution Research 23:18947-18951
  19. Storck, V., Karpouzas D.G., Martin-Laurent F (2017) Towards a new pesticide registration policy. Science of the Total Environment 575: 1027-1033
  20. Storck V., Nikolaki S., Perruchon C., Chabanis C., Sacchi A., Pertile G., Baguelin C., Karas P.A., Spor A., Devers M., Papadopoulou E.S., Sibourg O., Malandain C., Trevisan M., Ferrari F., Karpouzas D.G., Tsiamis G., Martin-Laurent F. (2018). Lab to field assessment of the ecotoxicological impact of chlorpyrifos, isoproturon or tebuconazole on the diversity and composition of the soil bacterial community. Frontiers in Microbiology 9:1412. doi: 10.3389/fmicb.2018.01412
  21. Karas P.A., Baguelin C., Pertile G., Papadopoulou E.S., Nikolaki S., Storck V., Ferrari F., Trevisan M., Ferrarini A., Fornasier F., Vasileiadis S., Tsiamis G., Martin-Laurent F., Karpouzas D.G., (2018) Assessment of the impact of three pesticides on microbial dynamics and functions in a lab-to-field experimental approach. Science of the Total Environment 637-638: 636-646
  22. El Azhari, N., Dermou E., Barnard R.L., Storck V., Tourna M., Beguet J., Karas P., Lucini L., Rouard N., Botteri L., Ferrari F., Trevisan M., Karpouzas D.G., Martin-Laurent F., (2018) The dissipation and microbial ecotoxicity of tebuconazole and its transformation products in soil under standard laboratory and simulated winter conditions. Science of the Total Environment 637-638: 892-906
  23. Vasileiadis S., Puglisi E., Papadopoulou E.S., Pertile G., Suciu N., Papolla, A., Tourna M.,  Karas P.A., Papadimitriou F., Kasiotakis A, Ipsilanti N., Ferrarini A., Sulowic S., Fornasier F., Nicol G.W, Trevisan M., Karpouzas D.G (2018) Blame it on the metabolite: 3,5-dichloraniline rather than the parent compound iprodione induces strong effects on the diversity and function of soil microorganisms. Applied and Environmental Microbiology 84(22): e01536-18
  24. Gallego-Blanco S., Devers-Lamrani M., Rousidou K., Karpouzas D.G., Martin-Laurent F., (2019) Assessment of the effects of oxamyl on the bacterial community of an agricultural soil exhibiting enhanced biodegradation. Science of the Total Environment 651: 1189-1198
  25. Suciu N., Vasileiadis S., Puglisi E., Pertile G., Tourna M., Karas P.Α., Papolla, A., Ferrarini A., Sulowic S., Fornasier F.,  Trevisan M., Karpouzas D.G., (2018) Azadirachtin and trifloxystrobin had no inhibitory effects on key soil microbial functions even at high dose rates. Applied Soil Ecology (in press)


- Dr. Evangelia Papadopoulou, Postdoctoral Fellow