TY - JOUR T1 - In vivo bioluminescent imaging reveals temporal and spatial dynamics of giardiasis JF - bioRxiv DO - 10.1101/080226 SP - 080226 AU - NR Barash AU - C Nosala AU - JK Pham AU - SG McInally AU - S Gourguechon AU - B McCarthy-Sinclair AU - SC Dawson Y1 - 2016/01/01 UR - http://biorxiv.org/content/early/2016/10/12/080226.abstract N2 - Giardia lamblia is a protistan parasite causing acute and chronic diarrheal disease in over one billion people worldwide, primarily in areas lacking adequate water treatment. Animal hosts ingest Giardia cysts that excyst in the gut to become motile trophozoites. Trophozoites colonize the small intestine, later differentiating into infectious cysts that are excreted and can contaminate water sources. Due to the limited accessibility of the gastrointestinal tract, our understanding of in vivo temporal and spatial dynamics of giardiasis is largely inferred from parasite physiology in laboratory culture. While in vitro models of giardiasis are informative, they may not adequately mirror in vivo parasite physiology in the host. To evaluate in vivo giardiasis directly, we developed bioluminescent imaging (BLI) methods to quantify temporal and spatial dynamics of giardiasis in mice using parasites expressing constitutive or encystation-specific luciferase bioreporters. Consistent with prior work, BLI confirms that metabolically active parasites primarily colonize the proximal small intestine. Contrasting with previous studies, we find that encystation is initiated and peaks early during infection and is localized to foci of high parasite density in the small intestine. Both BLI and immunostaining of encystation-specific vesicles (ESVs) corroborate that encystation is initiated in the proximal rather than the distal small intestine. This non-invasive method of imaging giardiasis provides an unprecedented and precise quantification of in vivo temporal and spatial patterns of infection, and an improved animal model for evaluation of anti-giardial drugs.Author Summary Giardia is a single-celled parasite causing both acute and chronic diarrheal disease in over one billion people worldwide. Direct quantification of Giardia infections in the host gut is difficult due to limited access to the site of infection. To evaluate parasite infections in living hosts, we developed in vivo imaging methods to quantify Giardia expressing bioluminescent physiological reporters in mice. Parasites primarily colonize the proximal small intestine in discrete, high-density foci. Encystation is also initiated at discrete foci in the small intestine early during infection, rather than in the colon as has been previously assumed. This work supports a model of a parasite density-based threshold for the induction of encystation in the proximal small intestine and in in vitro culture. ER -