Abstract
The evaluation of soil quality requires the use of robust methods to assess biologically based indicators. Among them, enzyme activities are used for several decades, but there is a clear need to update their measurement methods for routine use, in combining feasibility, accuracy, and reliability. To this end, the platform Biochem-Env optimized a miniaturized method to measure enzyme activities in soils using colorimetric substrates in micro-well plates. The standardization of the method was carried out within the framework of ISO/TC 190/SC 4/WG 4 “Soil quality – Biological methods” workgroup, recommending an inter-laboratory evaluation for the publication of a full ISO standard. That evaluation, managed by the platform, was based on the measurement, in six soils of contrasted physicochemical properties, of the ten soil enzyme activities described in the standard. Eight laboratories were involved in the validation study. Only 2.7% of outliers were identified from the analyses of the whole dataset. The repeatability and reproducibility of the method were determined by computing, respectively, the intra-laboratory (CVr,) and inter-laboratory (CVR) coefficients of variation for each soil and enzyme. The mean CVr ranged from 4.5% (unbuffered phosphatase) to 9.9% (α-glucosidase), illustrating a reduced variability of enzyme activities within laboratories. The mean CVR ranged from 13.8% (alkaline phosphatase) to 30.9% (unbuffered phosphatase). Despite this large CVR noticed for unbuffered phosphatase, the method was repeatable, reproducible, and sensitive. It also proved to be applicable for measuring enzyme activities in different types of soils. These results have been found successful by ISO/TC 190/SC4 and resulted in the publication of ISO 20130:2018 standard.
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All data, as well as supporting information, are currently available on https://data.inrae.fr/dataset.xhtml?persistentId=doi:10.15454/9ZQKK1.
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Acknowledgements
The authors would like to thank all the people who contributed to this project, particularly Sébastien Louis-Rose from AFNOR as manager of the committee T95E, and Antonio Bispo from INRAE (formerly at ADEME), for their important support and precious advices. Biochem-Env (https://doi.org/10.15454/HA6V6Y) is a service of the “Investissement d’Avenir” infrastructure AnaEE-France. The department ECOSYS is member of the LabEx BASC, funded by “Investissement d’Avenir” Program overseen by the French National Research Agency (ANR) (LabEx BASC, ANR-11-LABX-0034), and member of the Federation Ile-de-France of Research for the Environment (FR 3020 FIRE).
Funding
The study was funded by ADEME (Agence de l’Environnement et de la Maîtrise de l’Energie, grant N°1960C0096) in France. Catarina R. Marques was funded by national funds (OE), through FCT — Fundação para a Ciência e a Tecnologia — I.P., in the scope of the framework contract foreseen in the numbers 4, 5, and 6 of the article 23, of the Decree-Law 57/2016, of August 29, changed by Law 57/2017 of July 19. AnaEE-France is overseen by the French National Research Agency (ANR) (ANR-11-INBS-0001).
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Nathalie Cheviron, Virginie Grondin, and Christian Mougin were responsible for the design of the present study. Nathalie Cheviron and Christian Mougin prepared and wrote the original draft. Nathalie Cheviron and Virginie Grondin curated the data and performed the statistical analyses. All co-authors performed the experiments, acquired and provided the data, reviewed the manuscript, and approved the final version.
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Cheviron, N., Grondin, V., Marrauld, C. et al. Inter-laboratory validation of an ISO test method for measuring enzyme activities in soil samples using colorimetric substrates. Environ Sci Pollut Res 29, 29348–29357 (2022). https://doi.org/10.1007/s11356-021-17173-3
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DOI: https://doi.org/10.1007/s11356-021-17173-3