qRT-PCR analysis of bacteria copy numbers in feces of NOD mice shows a significant dysbiosis in mice on neutral drinking water. (A) qRT-PCR analysis of the total copy numbers of NOD mice on acidified (NOD-A), acidified to neutral (NOD-AtoN), or neutral water (NOD-N) drinking water (upper left) at pre-insulitis (5 weeks), post-insulitis, but before overt diabetes (10–13 weeks), and advanced progression of disease (16–20 weeks). Bacterial populations of Lactobacillus sp. (upper right), Bacteroides sp. (lower left), and C. coccoides (lower right) in the feces of female NOD-A (white, n=8–10), NOD-AtoN (striped, n=5), or NOD-N, black, n=8–10) mice. (B) Analysis of fecal microbial populations between 2-week-old NOD-A and NOD-N mice comparing numbers of total bacteria, Lactobacillus sp., Bacteroides sp., and C. coccoides (NOD-A, n=4, NOD-N, n=5). Significance was determined using Welches’ t-test at p<0.05. Significance is indicated by *- NOD-A vs NOD-N, †- NOD-A vs NOD-AtoN, and ‡- NOD-N vs NOD-AtoN.