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Lizards belonging to the genus <i>Anolis</i> are native to America and have been introduced in many parts of the world. In this work, a gastrointestinal microbiological analysis from <i>Anolis</i> sp. introduced to Tenerife, Canary Island, was carried out. A total of 74 individuals were analyzed by culture and molecular tools. <i>Pseudomonas</i> spp. was the most prevalent bacteria isolated (64.3%), followed by enteropathogenic <i>Escherichia coli</i> with at least one of the investigated virulent genes (<i>stx₁, stx₂,</i> and <i>eae</i>) (44.6%). The <i>stx₂</i> gene was more prevalent which differs to that reported in other reptiles, probably due to wastewater transmission. <i>Campylobacter</i> spp. was detected in 32.4% of the animals, highlighting the detection of <i>C. jejuni</i> and <i>C. fetus</i> by their relevance to public health. The zoonotic <i>Staphylococcus lugdunensis</i>, found in 14.9% of the animals, was firstly detected in reptiles. <i>Vibrio</i> sp. which is more associated with aquatic environments was found in 10.8% of the lizards in this study, with <i>Vibrio cholerae</i> being found in two of the animals. The prevalence of <i>Salmonella</i> sp. (5.4%) was low, compared with other studies carried out in reptiles. These results indicate that <i>Anolis</i> sp. in Tenerife could be playing a role in the maintenance and spread of the pathogens detected, being a possible risk factor for public health and biodiversity conservation.