Background: Oral diseases are common in dogs, with microbiota playing a prominent role in the disease process. Oral cavity habitats harbor unique microbiota populations that have relevance to health and disease. Despite their importance, the canine oral cavity microbial habitats have been poorly studied. The objectives of this study were to (1) characterize the oral microbiota of different habitats of dogs and (2) correlate oral health scores with bacterial taxa and identify what sites may be good options for understanding the role of microbiota in oral diseases. We used next-generation sequencing to characterize the salivary (SAL), subgingival (SUB), and supragingival (SUP) microbial habitats of 26 healthy adult female Beagle dogs (4.0 ± 1.2 year old) and identify taxa associated with periodontal disease indices.Results: Bacterial species richness was highest for SAL, moderate for SUB, and lowest for SUP samples (p < 0.001). Unweighted and weighted principal coordinates plots showed clustering by habitat, with SAL and SUP samples being the most different from one another. Bacteroidetes, Proteobacteria, Firmicutes, Fusobacteria, Actinobacteria, and Spirochaetes were the predominant phyla in all habitats.Paludibacter,Filifactor, Peptostreptococcus, Fusibacter, Anaerovorax, Fusobacterium, Leptotrichia, Desulfomicrobium, andTG5were enriched in SUB samples, whileActinomyces,Corynebacterium,Leucobacter,Euzebya,Capnocytophaga,Bergeyella,Lautropia, Lampropedia, Desulfobulbus, Enhydrobacter, andMoraxellawere enriched in SUP samples.Prevotella, SHD-231, Helcococcus, Treponema, andAcholeplasmawere enriched in SAL samples.p-75-a5,Arcobacter, andPasteurella were diminished in SUB samples.Porphyromonas,Peptococcus,Parvimonas, andCampylobacterwere diminished in SUP samples, whileTannerella,Proteocalla,Schwartzia, andNeisseriawere diminished in SAL samples. Actinomyces, Corynebacterium, Capnocytophaga,Leptotrichia, andNeisseriawere associated with higher oral health scores (worsened health) in plaque samples.Conclusions: Our results demonstrate the differences that exist among canine salivary, subgingival plaque and supragingival plaque habitats. Salivary samples do not require sedation and are easy to collect, but do not accurately represent the plaque populations that are most important to oral disease. PlaqueActinomyces,Corynebacterium,Capnocytophaga,Leptotrichia, andNeisseriawere associated with higher (worse) oral health scores. Future studies analyzing samples from progressive disease stages are needed to validate these results and understand the role of bacteria in periodontal disease development.