removed first) which can be sent to the laboratory for identification. Samples should be stored in a freezer if they are not processed within one or two days. The processing of the samples includes washing, drying, pinning, labelling and maintaining the specimens. Lastly, the data from the specimens should be entered into a database system, together with validation and double-checking procedures.
The species to be monitored should include both common (with regional-specific lists) and key indicator species, and cover different pollinator groups, including wild bees, hoverflies and butterflies.
In parallel, a monitoring scheme for the products of the beehive (pollen, nectar, honeybee bread and honey) could be rolled out to monitor pesticide residues and other environmental pollutants.
Soil represents a complex habitat sustaining a huge diversity of organisms that are structured by and embedded within the physical matrix (Geisen et al., 2019). Despite its importance for a range of ecosystems and ecosystem services, from nutrient cycle regulation to soil erosion control (Barrios, 2007), it is recognized that our knowledge of the soil habitat is limited (Jeffery et al., 2010). Currently, there are no specific policy measures or designated protection areas in the European Union targeting soil biodiversity. In a perspective of filling this gap, a pilot campaign was launched within the soil module of the 2018 ‘Land Use/Cover Area frame statistical Survey’ (LUCAS Soil), to test a sampling protocol for soil biodiversity. The campaign collected samples from 1 000 locations with diverse land cover and use. This is currently the most extensive EU assessment of soil biodiversity, based on DNA meta-barcoding (Orgiazzi et al., 2015).
The main aims of the survey are:
i to be able to develop a quantitative indicator of soil biodiversity, based on the genetic signatures,
ii to look for correlations between the DNA evidence and land cover/land use, especially in intensive agricultural areas,
iii to match the DNA data with residues of plant protection products. Information on the latter is available from a separate analysis of the concentrations of 70 active ingredients and metabolites in around 3000 LUCAS samples, and
iv eventually to expand the analysis to look at functional groups.
The DNA meta-barcode analysis will cover different types of soil-living organisms, from micro-organisms to macrofauna, and precisely (Orgiazzi et al., 2018):
• bacteria and archaea – target region 16S ribosomal DNA (rDNA),
• fungi – target region internal transcribed spacer (ITS), and
• eukaryotes other than fungi – target region 18S ribosomal DNA (rDNA).
Possibly, nematodes, arthropod mesofauna, and earthworms will be included. The protocol defined by the Earth Macrobiome Project (EMP, 2017) will be applied.
Soil samples should be frozen as soon as possible after collection. This requires precise logistical arrangements (Fernandez-Ugalde et al., 2017): the surveyors need to prepare freezer packs well in advance of the sampling and place them in a polystyrene box the day of the survey, wash the sampling material before each sampling with alcohol and water and wear plastic gloves during the collection of the sample. Once sealed in the polystyrene box, the sample, wherever the sampling point is located in Europe, should reach the final storage location preferably within 48 hours.
The reference for locating the sampling point is the LUCAS grid. Soil samples should be taken within a maximum of 100 m distance from the LUCAS point and in the same field where the LUCAS point is located. Surveyors use a metallic ring that they press in the soil with the help of a mallet to extract the soil sample. Ideally, a sample is taken at the LUCAS point, and mixed with four other samples collected 2 m away from the point in each of the main cardinal directions (Fig. 3). A subsample of soil is extracted and put in a plastic jar, labelled and sealed; the jar is put in the polystyrene box together with the freezer packs, labelled and sealed with tape.
Figure 3 Spatial distribution of samples to be collected for soil biodiversity analysis per each visited LUCAS point.
Initial results should be available during 2020. All information will be hosted by the European Soil Data Centre (ESDAC). This will include options for the online generation of maps as well as the entire methodology. Other parties (e.g. national organisations, NGOs) who wish to make the database grow through their own contribution may adopt the same protocol (Orgiazzi et al., 2018).
6Monitoring under the Habitats Directive 92/43/EEC
The Habitats Directive 92/43/EEC is aimed at maintaining or restoring ‘at favourable conservation status, natural habitats and species of wild fauna and flora of Community interest', which are listed in the Annexes of the Directive. Such requirements involve (i) defining ‘conservation status’; (ii) knowing where the habitat/species is found and (iii) what its conservation status is.
Conservation status is defined as ‘the overall assessment of the status of a habitat type or a species at the scale of a Member State’s biogeographical or marine region’ (DG Environment, 2017) and more precisely Article 1 of the Directive links the term to the extent of the area in which the habitat/species is found, the surface of the habitat area, its structure and functions (in case of habitat), the size of the population, its age structure, mortality and reproduction (of species) (EC, 2009).
Article 17 of the Habitats Directive (CEC, 1992) requires the Commission to draw up, every six years, a composite report based on the national reports delivered by the Member States of the European Union, including ‘in particular information concerning the conservation measures referred to in Article 6(1) as well as evaluation of the impact of those measures on the conservation status of the natural habitat types of Annex I and the species in Annex II and the main results of the surveillance referred to in Article 11’. The report should be available for the other EU institutions and the public in general.
Three of these reports are available (the fourth to be released in 2020), two of which focus on the conservation status of the habitat types and species included in the Annexes to the Directive for the periods 2001–2006 and 2007–2012. The assessment of conservation status takes as a reference the concept of favourable conservation status, defined as ‘a situation where a habitat type or species is prospering (in both quality and extent/population) and with good prospects to continue to do so in the future’, which is the overall objective of the Directive.
The conservation status of a species in the Habitats Directive (Article 1(i)) will be taken as ‘favourable’ when:
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