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In this paper, the mouse pup-in-a-cup model was improved for younger mouse pups, and the effect of artificial rearing on gut microbiota development was evaluated. Intragastric cannulas were placed through the esophagus into 3-day-old C57BL/6J mice (n=48), and the mice were artificially reared (AR) with mouse milk substitute (MMS). Littermate pups (n=20) were maternally reared (MR) as controls. The feces of 3-day-old pups were analyzed by combining the PCR-denaturing gradient gel electrophoresisdoi:10.1538/expanim.61.453 pmid:22850645 fatcat:4w3mty23kneaphkw5zx2drjj3a