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.

Central Institute for Supervising and Testing in Agriculture Brno Czech Republic

CESAM Centre of Marine and Environmental Studies Department of Biology University of Aveiro Campus Universitário de Santiago 3810 193 Aveiro Portugal

Departamento de Microbiología 3 Facultad de Biología Universidad Complutense de Madrid 28040 Madrid España

Dpto de Química en Ciencias Farmacéuticas UD Edafología CARESOIL Facultad de Farmacia Universidad Complutense de Madrid 28040 Madrid España

INERIS Direction des Risques ChroniquesUnité EXES F 60550 Verneuil en Halatte France

INRAE UMR Eco and Sols F 34060 Montpellier France

UniLaSalle Campus Rouen AGHYLE Research Unit UP 2018 C101 F 76134 Mont Saint Aignan France

Université Paris Saclay INRAE AgroParisTech UMR ECOSYS F 78026 Versailles France

Université Paris Saclay INRAE Plateforme Biochem Env F 78026 Versailles France

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