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This work investigated the genetic relationship among Stenotrophomonas maltophilia strains in fecal samples from dairy cows in northeast China and identified the dominant β-lactamase genotype. One hundred and six samples were collected from two randomly selected cow farms in northeast China, and the isolates were identified with MALDI-TOF/MS. Whole-genome sequencing was conducted using Illumina HiSeq 4000-PE150 platform (Illumina, Inc., USA). The antimicrobial resistance genes were detected using CGE services. The phylogenetic analysis of S. maltophilia strains was performed by Roary and MEGA X. In total, 24 S. maltophilia isolates were isolated. The results of resistome analysis showed all S. maltophilia strains carrying blaL1 gene, which was the only β-lactamase genotype. In addition, the aminoglycoside resistance genes aac(6′)-Iz and aph(3′)-IIc were found. The phylogenetic tree indicated the clonal diversity of S. maltophilia in these two regions and the clonal relatedness of the strains from these regions. This study first investigated the dissemination and characterization of S. maltophilia isolates from dairy cows in northeast China and provided evidence of the potential transmission between two provinces. Furthermore, it indicated blaL1 was the most prevalent genotype of β-lactamase in these regions.