High-density foci of Giardia colonization are present in the proximal small intestine of mice and gerbils. High-density foci of Giardia colonization are present in the proximal small intestine of mice and gerbils. (A) Representative classes of in vivo and ex vivo bioluminescent images are shown for 24 mice infected with the PGDH-FLuc strain and sacrificed in cohorts of four on days 3, 5, 7, 9, 11, and 13 postinfection. Photon flux or radiance (photons/second/square centimeter/steradian) for each intestinal segment is shown and has been normalized to the maximal ex vivo bioluminescence signal on the radiance scale, yielding the percent total signal per segment. These values are represented graphically on the grayscale maps below each ex vivo image (white, 0 to 10%; black, 75 to 100%; values between 10% and 75% indicated as shades of gray). A strikethrough indicates greater than 90% of maximal bioluminescent signal. P, proximal small intestine; D, distal small intestine; C, cecum; L, large intestine. The stomach (S) is shown for orientation but always lacks bioluminescence. (B) Quantitative bioluminescence imaging from infections is categorized and summarized by the region of the gastrointestinal tract for all animals in each phase of infection as early (days 0 to 3), middle (days 5 to 9), and late (days 11 to 13). Shading in each row of the charts indicates the variations in the maximal bioluminescence in each of the four regions in an individual animal. (C) Degree to which the in vivo PGDH-FLuc bioluminescence is significantly and linearly correlated in 24 intestinal samples from four infected mice with quantitative PCR measures of Giardia abundance in the same samples using a single-copy Giardia gene (PFOR1) (normalized to the mouse single-copy nidogen gene). Statistical significance (R = 0.9157, P < 0.0001) is noted. (D) The same PGDH-FLuc strain was used to infect two Mongolian gerbils (see Materials and Methods) without antibiotic pretreatment, and animals were noninvasively imaged over 15 days. Day 8 p.i. and day 15 p.i. noninvasive whole-animal imaging and ex vivo gastrointestinal tract imaging for day 15 p.i. are shown for both animals (1 and 2) with anatomical annotations as in panels A and B. N. R. Barash et al. mSphere 2017; doi:10.1128/mSphere.00343-16