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All mice were fed standard rodent chow (PicoLab 5053, LabDiet) and housed in a specific pathogen-free facility at a controlled temperature of 2223 C on a 12-h lightdark cycle

10.1016/j.reprotox.2021.01.003 57 JostA

Four replicates from each group were analyzed independently for library synthesis and sequencing

Although the exact mechanism of how coffee enemas cause proctocolitis is not known, it is suspected that the proctocolitis is caused by the chemicals in coffee enemas, as coffee is a complex mixture of chemical compounds that contain more than just caffeine