Camel milk has significant economic value and is an important food in the region of Alxa Left Banner of Inner Mongolia. Fifteen fresh camel milk samples were collected from domesticated camels in a pasture of Alxa Left Banner. The physicochemical properties and bacterial diversity of camel milk samples were analyzed. The average values of fat, total protein, nonfat milk solids, acidity, and density were 4.40%, 3.87%, 9.50%, 16.95°T, and 1.02 g/cm3, respectively. The bacterial microbiota of the collected fresh camel milk was investigated using PacBio single-molecule real-time (Pacific Biosciences, Menlo Park, CA) sequencing. The camel milk microbiota was highly diverse and comprised 8,513 operational taxonomic units belonging to 32 phyla, 377 genera, and 652 species. The major phyla included Proteobacteria, Firmicutes, Deinococcus-Thermus, Bacteroidetes, and Actinobacteria. A small number of lactic acid bacteria sequences were detected, representing the species Streptococcus thermophilus, Lactobacillus helveticus, Lactococcus lactis, and Leuconostoc mesenteroides. A total of 72 strains of lactic acid bacteria were isolated and identified from 15 samples, including Lactobacillus paracasei, Enterococcus italicus, Enterococcus durans, Lactococcus lactis ssp. lactis, Weissella confusa, and Enterococcus faecium. These results confirm that fresh camel milk has a high bacterial diversity and is a valuable natural resource for isolation of novel lactic acid bacteria. Copyright © 2020 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.