Background Most of the Basque Country is considered as “disfavoured and mountain agriculture area”, according to the European Directive 75/268/CEE. In mountain areas, the primary activity is restricted by the physical limitations of the environment. Traditional farming activities are based on transhumance – the movement of livestock to higher pastures in summer and to lower valleys in winter. The socio-economic value of the grazing areas has been increased by the inclusion of many of these habitats in Natura 2000 network sites. These habitats include mountain grasslands, Alpine and subalpine calcareous grasslands, and European dry heaths. This project focuses on the Gorbeia Natural Park and its vicinity that hosts a range of grazing habitats and consolidated livestock farming. Management of these habitats has been mainly based on traditional approaches using visible parameters, especially levels and types of vegetation. But these practices are not the best way of conserving soil biodiversity. New methodologies offer more efficient management of the mountainous areas. Objectives The main objective of the PRO-Izki project was to demonstrate the viability of an innovative methodology for the conservation of soil and vegetation biodiversity in mountain and bottom valley grazing areas, based on the application of Agroecosystems’ Health Cards (TSA). This tool would demonstrate the positive impact of certain agricultural operations (existing or alternative ones) in the conservation of soil and vegetal biodiversity in several grazing habitats. The project will propose soil (micro)biological indicators for the integral diagnosis of the health of the grazing agroecosystems together with the traditional physiochemical indicators. A portable near infrared spectroscopy (NIR) technology would allow the prediction of the soil health to be enlarged to broader areas in a quick and affordable way. It was hoped that this would demonstrate the relationship between the biodiversity on the surface (vegetation) and under the surface (soil) in grazing agroecosystems, as well as advance knowledge of the mechanisms of such relationships. At the same time, the project intended to give value to pasture for the conservation of vegetal and soil biodiversity in grazing agroecosystems through a set of awareness-raising activities. Results of the project would be disseminated to ensure the transference and easy replicability of the developed methodology both at national and at European level. Results The PRO-Izki project implemented several farming practices over 120 ha of pasture land with the aim of preserving and boosting these agroecosystems and the biodiversity they host. In the case of the mountain areas, the project demonstrated the feasibility of using the herbicide Asulox to control the expansion of the bracken fern (Pteridium aquilinum). In lower altitudes, fertilisation with organic fertilisers (i.e. manure) yielded a similar production rate to that obtained by mineral fertilisation. Liming was shown to have a positive impact on the species with high forage value, such as Lolium perenne. The project moreover acquired valuable data, previously unknown, on the biodiversity of the Atlantic grasslands of the Iberian Peninsula, especially their soil biodiversity. It furthermore identified the soil microbial diversity at species level of the grasslands thanks to pyrosequencing analysis. The project team, technicians and researchers of NEIKER, worked closely with the Basque Government and the Provincial Council of Bizkaia, and technicians and farmers of the Lorra Cooperative and the Farmer Associations of Orozko and Zeanury, to develop the technical actions foreseen, as well as the management and dissemination of the project and the use of the TSA. The first task was to design and establish a georeferenced environmental network that compiled the diversity of the most representative pasture areas and farms of the Gorbeia National Park and its surroundings. This diversity refers to both ecological factors (type of vegetation, altitude, geological material, orientation, etc.) and to factors linked to livestock management (livestock density, type of animals, breeds and presence of livestock infrastructure, among others). The project then focused on the design of the TSA – i.e. manuals that explain in a simple and practical way how to access the health status of different pastoral ecosystems. The TSA thus allows managers of the agroecosystems to take decisions about the farming practices that should be boosted based on objective criteria. TSAs, which until now have not been widely used in Europe, could also be a useful instrument at European level, particularly in regards to the Common Agricultural Policy. Other ongoing projects and initiatives are incorporating the TSA in cooperation with the beneficiary. Furthermore, the TSA allows even those without any previous training to decide on the most suitable farming practices for their particular pedo-climatic conditions and agroecosystems. It leads therefore to more sustainable and profitable farms, given that improved soil will give rise to higher quality pasture. It is also an affordable approach since farmers do not have to invest in costly equipment. The farming practices were implemented in 33 different pasture areas. Actions included: Clearing of bushes (40 ha) in habitat 4030; Control of ferns (4 ha) with a herbicide and through brush cutting; Fertilisation of mountain habitats (60 ha) by applying phosphorus; Manure and mineral fertilisation of valley habitats (10 ha); Liming in the valley (8 ha) by applying Ca(OH)2 vs. non-application. Pastures for harvesting, grazing and mixed; and Fencing of pastures in 16 areas (four for each habitat targeted).Due to the bad weather conditions during the autumns of 2012 and 2013, the application of mineral fertilisers had to be postponed until the springs of 2013 and 2014 in order to avoid the loss of nutrients. This delay nevertheless highlighted the need to consider the risk of nutrient leaching. Applying the TSA, the project was able to assess the impact of these practices on soil and vegetation. The project team analysed soil microbial indicators, along with the more traditional physicochemical indicators. Soil microorganisms were sequenced and identified using pioneering next generation sequencing technologies (among other analyses). Analysis also took into account parameters such as plant diversity and grass production.