Unraveling the Risk Factors and Etiology of the Canine Oral Mucosal Melanoma: Results of an Epidemiological Questionnaire, Oral Microbiome Analysis and Investigation of Papillomavirus Infection

SIMPLE SUMMARY: Oral mucosal melanoma (OMM) is one of the most common oral cancers in dogs; however, the risk factors for its development remain obscure and the etiology is unknown. This study aimed to investigate the risk factors and etiology of OMM in dogs. An epidemiological questionnaire was applied to the owners of 15 dogs with OMM and their paired controls, and the oral microbiome was comparatively determined in the two groups. Additionally, the presence of papillomavirus was investigated in the same OMM samples. Most OMM and control dogs had grade 3 periodontal disease. No risk factors were identified through the epidemiological questionnaire, and papillomaviruses were not identified in the samples. The bacteria Tannerella forsythia and Porphyromonas gingivalis were significantly overrepresented in dogs with OMM, and their presence could be considered a risk factor for the development of canine OMM. ABSTRACT: Oral mucosal melanoma (OMM) is the most common oral cancer in dogs and is very aggressive in this species; its risk factors and etiology are yet to be determined. This study aimed to unravel the risk factors for the development of OMM in dogs and to investigate the possible presence of papillomaviruses as an etiological factor. A case-control study was conducted in 15 dogs with OMM and 15 paired controls whose owners answered an epidemiological questionnaire. Oral swabs from the same dogs were subjected to 16S rRNA sequencing for microbiome analyses. In addition, DNA fragments of OMM had their DNA extracted and amplified by polymerase chain reaction in an attempt to detect canine papillomaviruses. The gingiva was the most frequent anatomical site (47%) of OMM, and most tumors were stage III when diagnosed. Most dogs bearing OMM and the controls had grade 3 periodontal disease, and this factor, along with tartar treatment and tooth brushing, did not differ between cases and controls. Most dogs with OMM and most controls had contact with smokers; there was no statistically significant difference. Canine papillomaviruses were not detected among OMM cases. Tannerella forsythia and Porphyromonas gingivalis were significantly increased in case dogs compared to the controls. As these bacteria are reportedly involved in the pathogenesis of periodontal disease and esophageal cancer in humans, we suggest that they might be risk factors for the development of canine OMM. The limitations of this study include the low number of dogs, and therefore, further studies on canine OMM with larger numbers of animals are encouraged.