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However, it should be emphasized that use of caloric restriction to control for weight-dependent effects in DIO mouse studies may potentially introduce confounding effects on microbiome signatures due to significant compensatory changes in hunger sensation, feeding pattern, resting metabolic rate and gut morphology which may be disproportionately to that attained by appetite suppressants 80,81

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And even if we use sterile water for injection (WFI, water for injection ), we run into another problem: as soon as you pierce the rubber stopper of the vial with a needle and withdraw part of the solution, bacteria from the air can enter the remainder through the same opening