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Strains of <i>Pseudomonas savastanoi</i> pv. <i>savastanoi</i> (<i>Pss</i>), isolated from infected olive trees (<i>Olea europaea</i> L.) in three European countries (Croatia, Slovenia and Portugal) were identified and characterised according to their colony morphology, physiological and biochemical features. According to the LOPAT scheme, 38.6% of <i>Pss</i> isolates were grouped in the Ib cluster. The Portuguese <i>Pss</i> strains were fully consistent with the typical LOPAT profile for this bacterium. Conversely, most Slovenian <i>Pss</i> strains showed delayed oxidase activity, whilst Croatian <i>Pss</i> strains did not produce any fluorescent pigment when grown in vitro. For <i>Pss</i> molecular identification, both end-point and real-time PCR were used, as well as MALDI–TOF, which was additionally used for proteomic analysis and the subsequent species identification of a number of strains that showed deviations from expected LOPAT results. <i>Pss</i> was confirmed as a causal agent of olive knot disease in 46.6% of olive orchards screened. Overall, these data suggests a possible correlation of certain <i>Pss</i> features with the geographical origin and the ecological niche of <i>Pss</i> isolates.