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Wildlife can harbour Shiga toxin-producing <i>Escherichia coli</i> (STEC). In the present study, STEC in faecal samples from red deer (<i>n</i> = 106) and roe deer (<i>n</i> = 95) were characterised. All isolates were non-O157 strains. In red deer, STEC were detected in 17.9% (<i>n</i> = 19) of the isolates, and the <i>eae</i>/<i>stx</i>_2b virulence profile was detected in two isolates (10.5%). One STEC strain harboured <i>stx</i>_1a (5.3%) and eighteen STEC strains harboured <i>stx₂</i> (94.7%). The most prevalent <i>stx</i>₂ subtypes were <i>stx</i>_2b (<i>n</i> = 12; 66.7%), <i>stx</i>_2a (<i>n</i> = 3; 16.7%), and <i>stx</i>_2g (<i>n</i> = 2; 11.1%). One isolate could not be subtyped (NS) with the applied primers (5.6%). The most widely identified serotypes were O146:H28 (<i>n</i> = 4; 21%), O146:HNM (<i>n</i> = 2; 10.5%), O103:H7 (<i>n</i> = 1; 5.3%), O103:H21 (<i>n</i> = 1; 5.3%), and O45:HNM (<i>n</i> = 1; 5.3%). In roe deer, STEC were detected in 16.8% (<i>n</i> = 16) of the isolates, and the <i>eae</i>/<i>stx</i>_2b virulence profile was detected in one isolate (6.3%). Two STEC strains harboured <i>stx</i>_1a (12.5%), one strain harboured <i>stx</i>_1NS/<i>stx</i>_2b (6.3%), and thirteen strains harboured <i>stx₂</i> (81.3%). The most common subtypes were <i>stx</i>_2b (<i>n</i> = 8; 61.5%), <i>stx</i>_2g (<i>n</i> = 2; 15.4%), non-typeable subtypes (NS) (<i>n</i> = 2; 15.4%), and <i>stx</i>_2a (<i>n</i> = 1; 7.7%). Serotype O146:H28 (<i>n</i> = 5; 31.3%) was identified. The study demonstrated that the zoonotic potential of STEC strains isolated from wildlife faeces should be monitored in the context of the ‘One Health’ approach which links human health with animal and environmental health.