To evaluate the presence of thermotolerant Campylobacter spp. in stray cats in southern Italy.One hundred and thirteen animals were trapped in two different environments (urban area, harbour area) of the city of Naples. From each cat, rectal swabs were collected. The samples were processed in order to detect thermotolerant Campylobacter spp. by culture methods. The positive samples were then confirmed by multiplex polymerase chain reaction. Campylobacter jejuni was isolated from 19/113 (16.8%) stray cats, whereas Campylobacter coli was not isolated. The cat data (age, environment and sex) were analysed by two statistical analyses using the C. jejuni status (positive/negative) as a dependent variable. As regards statistical regression model results, age and environment were risk factors for C. jejuni positivity. In particular, cats older than 1 year had a significantly higher risk of being positive for C. jejuni than cats aged up to 1 year (OR = 10.440; P = 0.000). Moreover, cats living in the harbour area had a significantly higher risk to be a carrier of C. jejuni than cats living in urban area (OR = 17.911; P = 0.008).The findings of the present survey confirm stray cats as potential carriers of C. jejuni.This is the first study on the prevalence of C. jejuni in stray cats in Europe.