Background Large carnivores such as wolves and bears are often perceived as a problem by people living in or making a living in areas with these animals, who see them as threats to themselves or their livelihoods. However, large carnivores - as well as scavenger raptors such as the griffon vulture (Gyps fulvus) and bearded vulture (Gypaetus barbatus) - are under serious threat, especially from poisoning, even in protected areas. Objectives The overall aim of the ANTIDOTO project was to achieve effective preservation of large carnivores - wolves and bears - and of various species of scavenger raptors, in protected areas of Italy and Spain. The large carnivores in the project areas have already benefited from the activities of an earlier LIFE project, COEX (LIFE04 NAT/IT/000144), which raised awareness to promote a more positive coexistence between local stakeholders and these animals in five European countries, including Italy and Spain. The ANTIDOTO project sought to develop an integrated strategy covering reduced risk of poisoning, mitigating the conflict between large carnivores and farmers, and restocking key species in target areas. The project planned to establish and manage Anti-poison Dog Units to directly oversee reductions in the use of poison baits against large carnivores and raptors in the Gran Sasso and Monti della Laga National Park in Abruzzo (Italy) and in Aragon (Spain), and to encourage Italian national bodies to adopt measures to prevent such poisoning. Results The ANTIDOTO project implementation an innovative anti-poisoning strategy and methodology to help protect wolves, bears and scavenger raptor species from illegal poison baiting in protected areas in Italy and Spain. The methodology centred on the establishment of Anti-poison Dog Units, including the training of personnel and capacity building within public bodies. This methodology, involving dogs trained to detect specific bait poisons, was based on positive experience gained in Andalusia (Spain) through the involvement of associated beneficiary AMAYA. In Italy, the project set up two Anti-poison Dog Units (instead of one, as originally planned). This ensured a constant availability of the Anti-poison Dog Units on the ground, with rapid intervention capacity; and the ability to carry out timely inspections within, but also outside, the Gran Sasso and Monti della Laga National Park in response to requests from public bodies. The 202 inspections carried out by the Anti-poison Dog Units in the Gran Sasso and Monti della Laga National Park during the project did not find any poisoned baits, which in part was due to the awareness-raising campaign run by the project. However, outside the Park, the Anti-poison Dog Units carried out 69 urgent inspections, 31 of which in were in protected areas characterised by recurrent episodes of poisoning against grey wolf (Canis lupus), with 14 inspections finding poisoned baits and carcasses. The project supported the creation of other Anti-poison Dog Units beyond the project areas, namely three dogs were trained by LIFE project staff to find poisoned baits in Tuscany and Emilia-Romagna (Italy). Two field training courses were organised and attended by 28 people involved with the monitoring units (e.g. the Italian State Forestry Service (CFS) and Spanish law enforcement officers from SEPRONA). In Aragon (Spain), the Anti-poison Dog Unit established by the project team carried out 156 inspections, 56 of which found evidence of poisoning in carcasses and scattered baits. Seven inspections outside of the region did not find evidence of poisoning, but they were a good means of disseminating information about Anti-poison Dog Units. The project conducted advanced training activities on strategies to prevent and control illegal poisoning in Gran Sasso and Monti della Laga National Park, and in Andalusia and Aragon, through seven courses targeted to specific categories of stakeholders (e.g. forestry staff, private and public veterinarians, technical officers of public administrations). In total, 350 people participated in these training initiatives, which enhanced technical knowledge and awareness on illegal poisoning in protected areas. The project team published a technical manual on diagnostic methods for identifying poisoning, and diagnostic tests were conducted on poisoned animals (mainly wild and domestic dogs) in Italy and Spain. Awareness-raising activities were also targeted at livestock breeders, hunters, tourist operators, schoolchildren and the general public, both in Italy and Spain. The survey carried out to monitor these activities in Italy concluded that, among other things, 89% of the interviewed sample deemed the entire project action useful/very useful. The project improved conditions for the conservation of wolf and bear populations in Italy, and of scavenger raptors in Italy and Spain. The monitoring activities of the Anti-poison Dog Units revealed that illegal poison use in the respective territories has been reduced, proving that conflicts between wildlife and stakeholders have been mitigated. The project was also able to prevent the illegal killing of domestic animals in some urban areas surroundings the target natural areas. The strategy against illegal poisoning devised by the project included a draft for a national law to ban the use of poisonous baits, as a contribution to the development of Italian environmental policy and legislation. ANTIDOTO organised a national workshop ‘Strategy against the use of poison in Italy’, from which a report setting out the strategy was published. The use of small dog units trained to retrieve poisoned baits, over large areas of rough terrain, represents an innovative methodology to prevent illegal poisoning of wildlife. Following ANTIDOTO, numerous initiatives tackling the illegal use of poison baits have flourished all over Italy, such as training courses, conferences, and dedicated actions within ongoing projects. The project activities will continue in the Gran Sasso and Monti della Laga National Park, thanks to funds made available by the Italian Ministry of Environment, Land and Sea. In addition, the National Park is now the coordinating beneficiary of LIFE PLUTO (LIFE13 NAT/IT/000311), which started on 1 June 2014, with the CFS as an associated beneficiary. LIFE PLUTO will extend the ANTIDOTO experience throughout central and south Italy, by setting up six new Anti-poison Dog Units. ANTIDOTO’s strategy is also being adopted by the LIFE WOLFALPS (LIFE12 NAT/IT/000807), LIFE MEDWOLF (LIFE11 NAT/IT/000069) and LIFE MIRCO-lupo (LIFE13 NAT/IT/000728) projects that have set up Anti-poison Dog Units to operate in the Alpine and north Apennines regions, as well as in Mediterranean-type areas. In Spain, the ANTIDOTO project associated beneficiary AMAYA in Andalucia will continue to supply dogs trained to detect poison baits and to disseminate the project’s results to its staff and relevant stakeholders. The Government of Aragon will keep operating its Anti-poison Dog Units with these dogs and carry out forensic analyses to monitor illegal animal poisoning, together with the information dissemination campaign. Further information on the project can be found in the project's layman report and After-LIFE Conservation Plan (see "Read more" section).