Linear infrastructure represent a barrier to movement for many species, reducing the connectivity of the landscapes in which they reside. Of all linear infrastructure, roads and fences are two of the most ubiquitous, and are understood to reduce landscape connectivity for wildlife. However, what is often neglected consideration is a holistic approach of modelling the effects of multiple types of linear infrastructure simultaneously. Few studies have examined this, typically assessing the impacts of a singular kind of infrastructure on landscape connectivity. Therefore, the aim of this study is to address the relative importance of considering multiple kinds of linear infrastructure in landscape connectivity modelling. We utilised presence data of red deer Cervus elaphus and wild boar Sus scrofa in Doñana Biosphere Reserve (Spain) to generate a sequential approach of scenarios of landscape connectivity; firstly only with environmental variables, secondly with roads as the sole infrastructure, thirdly with the addition of fences, and finally with the further addition of fences and wildlife road-crossing structures. We found that the connectivity of the landscape was greatly affected by the addition of fences and wildlife road-crossing structures in both species, with fences in particular causing considerable alterations to estimated movement pathways. Our finding impresses a need to consider multiple different types of linear infrastructure when modelling landscape connectivity to enable a more realistic view of wildlife movement and inform mitigation and conservation measures more accurately., HORIZON EUROPE European Innovation Council : 101060429, European Union’s Horizon Europe Research and Innovation Programme; Government of the United Kingdom , Turing Scheme; Fundação para a Ciência e Tecnologia : 2022.01773.CEECIND, CEECIND; European Commission : LIFEWATCH-2019-09-CSIC-13; European Commission : POPE2014-2020; Ministry of Economy, Industry and Competitiveness : RYC2021-033192-I, Ramón y Cajal Program; Ministerio de Ciencia e Innovación : IJC2019-039662-I, Juan de la Cierva Incorporación Program, Peer reviewed

A list of articles on global change drivers in Doñana Protected Area has been provided as supporting information, Maria Paniw was supported by the grants RYC2021-033192-I and PID-2022-141004OA-100; Irene Mendoza by the grants PID2020-115129RJI00 and TED2021-129871A-I00 by MCIN/AEI/10.13039/501100011033 by “European Union NextGenerationEU/PRTR”; This project has received funding from the European Union's Horizon 2021 research and innovation program under the Marie Skłodowska-Curie grant agreement No 101064340 to Virginia Domínguez-García. María José Ruiz-López was supported by the grant PID2020-118921RJ-I00; Javier Puy by grant FJC2020-042954-I by MCIN/AEI/10.13039/501100011033 by “European Union NextGenerationEU/PRTR”; Laetitia M. Navarro, Alberto García-Rodríguez, Elena Tena, José M. Reyes-González, Simone Guareschi, Pablo Homet, Pablo Villalva and Eva Moracho were funded by the Spanish Ministry of Science and Innovation project through European Regional Development Fund (SUMHAL, LIFEWATCH-2019-09-CSIC-4, POPE 2014–2020). Cosme López-Calderón was supported by Juan de la Cierva fellowship (FJC2019-042541-I). Elena Velado-Alonso received funding from the project SHOWCASE (SHOWCASing synergies between agriculture, biodiversity and ecosystem services to help farmers capitalizing on native biodiversity) within the European Union's Horizon 2020 Research and Innovation Programme under grant agreement No. 862480. Cristina Coccia acknowledges the support of NBFC to Roma Tre University, under the National Recovery and Resilience Plan (NRRP), Mission 4 Component 2 Investment 1.4—Call for Tender No. 3138 of 16 December 2021, rectified by Decree No. 3175 of 18 December 2021 of Italian Ministry of University and Research funded by the European Union—NextGenerationEU. Project Code CN00000033, Concession Decree No. 1034 of 17 June 2022 adopted by the Italian Ministry of University and Research, CUP F83C22000730006. Alberto García-Rodríguez, Elena Tena, and Iñigo Donázar-Aramendía were additionally supported by the EU-NextGenerationEU funds through the 2021–2023 Margarita Salas call for the requalification of the Spanish university system, convened by the Universidad de León, Universidad de Salamanca, and Universidad de Sevilla, respectively. Sebastiano de Franciscis was funded by the Plan Andaluz de Investigación, Desarrollo e Innovación (PAIDI 2020) of Junta de Andalucía (PY20\_00765) granted to M.A.F. Zulima Tablado was supported by the NaturaConnect project within the Horizon Europe funding scheme (Grant agreement ID: 101060429). Marcello D'Amico was supported by the Juan de la Cierva Incorporación Program from the Spanish Ministry of Science, Innovation and Universities (IJC2019-039662-I), and the project “Plan Complementario de I+D+i en el área de Biodiversidad (PCBIO)” funded by the European Union within the framework of the Recovery, Transformation and Resilience Plan—NextGenerationEU and by the Regional Government of Andalusia, This work contributes to the Institut de Ciències del Mar "Severo Ochoa Centre of Excellence" accreditation CEX2024-001494-S funded by AEI 10.13039/501100011033 of the Spanish Ministry of Science and Innovation, Peer reviewed