AbstractDespite public concern on the role of free‐roaming cats as reservoirs of zoonotic agents, little is known about the influence of urban and peri‐urban landscapes on the exposure risk. We evaluated the seroprevalence of three zoonotic agents (Chlamydia felis, Coxiella burnetiiandToxoplasma gondii) in domestic cats (Felis catus). Two hundred and ninety‐one free‐roaming cats were trapped in Murcia municipality (Southeast Spain), and their sera were tested for specific antibodies againstT. gondiiusing a modified agglutination test (MAT), and forC. felis,C. burnetiiand feline immunodeficiency virus (FIV) antibodies with ELISA technique. Pathogen seroprevalence at 95% CI was calculated for each sex and age category (up to and over 12 months) and compared with a chi‐squared test. The role of human population density and urban landscape characteristics on the risk of pathogen exposure in the cat population was explored using generalized linear models. Seropositivity against a single pathogen was found in 60% of the cats, while 19% was seropositive for two or three pathogens. Seroprevalence ofC. feliswas 8% (CI95%: 5–11), 37% (CI95%: 31–42) forC. burnetiiand 42% (CI95%: 36–47) forT. gondii. In addition to these three pathogens, FIV seropositivity was low (1%, CI95%: −0.1 to 2) and adult cats were more likely to be seropositive toC. burnetiithan young individuals (OR: 2.3, CI95%: 1.2–4.2). No sex or age class differences in seroprevalence were observed for the rest of the pathogens. Seropositivity was correlated with water surface areas forC. felis,and not with crop areas.Coxiella burnetiiseropositivity was correlated with the percentage of urban areas (continuous with only buildings and discontinuous, that include buildings, parks, and pedestrian and urban green areas), human population size and peri‐urban areas with shrubs, and not correlated with other agricultural landscapes (orchards and crop areas). However, the seroprevalence ofT. gondiiwas only associated with agricultural landscapes such as orchards. The detection of hotspot areas of high pathogen exposure risk is the basis for municipal services to implement surveillance and risk factor control campaigns in specific‐risk areas, including (a) efficient health management of urban cat colonies by geographical location, population census and health status monitoring of the components of each cat colony, (b) improvement of hygiene and sanitary conditions at the feeding points of the cat colony and (c) free‐roaming cat trapping for health monitoring and, in the long term, to know the evolution of the health status of their populations.