ISO 11268-3:2014

Overview - ISO 11268-3:2014 (Soil quality, earthworms, field guidance)

ISO 11268-3:2014 provides guidance for determining the effects of pollutants and chemicals on earthworms in field situations. It specifies field techniques for assessing changes in earthworm species, abundance and biomass after substances are applied to or incorporated into soil (including soil injections and drilled pelleted formulations). The standard is intended to support ecotoxicology studies, environmental monitoring, and regulatory risk assessment for soil-applied chemicals such as pesticides.

Key topics and technical requirements

• Test principle: Compare species, numbers and biomass from treated plots with control and reference plots using a randomized complete block design (typically four replicates per treatment).
• Sampling methods: Combination of hand-sorting and formalin extraction (see ISO 23611-1). Sampling should occur during active periods for earthworms to avoid diapause.
• Plot guidance: Typical individual sample squares (e.g., 50 cm × 50 cm); adjust plot size by earthworm density (examples in the standard: 0.125 m2 for high density, 1 m2 for low density).
• Duration and timing: Study duration depends on the test substance but is usually about one year. Sampling dates are selected to match earthworm activity periods.
• Endpoints and analysis: Primary endpoints are abundance, biomass and species diversity. Statistical analysis of replicate samples is used to detect treatment effects.
• Units and reporting: Application rates in kg/ha or l/ha; soil-incorporated concentrations in mg active ingredient (a.i.)/kg soil dry mass.
• Reagents and apparatus: Lists commonly used reagents (e.g., formalin, ethanol) and basic field/lab equipment for collection, preservation and biomass determination.
• Test site selection: Prefer homogeneous grassland (recommended minimum densities specified); avoid heterogeneous sites (e.g., orchards) unless compensated by sampling design.
• Quality elements: Pre-sampling to confirm species distributions and densities, use of reference substances (carbendazim example), preservation and biomass protocols per referenced standards.

Applications and who uses ISO 11268-3:2014

• Environmental risk assessors and regulatory agencies evaluating soil ecotoxicity of pesticides and other chemicals.
• Pesticide manufacturers and registrants conducting required field studies for product registration.
• Soil ecologists, contract ecotoxicology labs, and environmental consultants performing field monitoring of soil quality and earthworm populations.
• Research institutions studying pollutant impacts on soil biodiversity and ecosystem services.

Related standards

• ISO 23611-1 - Soil sampling and earthworm extraction (hand-sorting, formalin).
• ISO 10390 - Soil pH determination.
• ISO 10694 - Soil organic/total carbon (dry combustion).
• ISO 11274 / ISO 11277 - Soil water-retention and particle size methods.
  Note: ISO 11268 series also includes Part 1 (acute toxicity) and Part 2 (reproduction tests).

Keywords: ISO 11268-3:2014, soil quality, earthworms, field test, ecotoxicology, pesticide field study, sampling, formalin extraction, hand-sorting, biomass, species diversity.