Project description:To investigate the transcriptional responses of intestinal epithelial cells and Giardia intestinalis, assemblage A isolate WB-C6, trophozoites during infection, we infected human enteroids with preconditioned trophozoites for 1h and 3h. Giardia intestinalis trophozoites were preconditioned before the infection with either DMEM/F-12 or DMEM/F-12 supplemented with 10% FBS to modify the trophozoites’ fitness.
Project description:The mechanisms underlying the coexistence of crAss-like phages and other types of phages infecting Bacteroidales are not well understood. This study examines potential mechanisms that promote phage coexistence in the Bacteroidales order through RNA-seq and proteomic analysis. The analysis was performed on anaerobic bacterial cultures co-cultured with phages over a period of 5 days and corresponding control cultures without phage, with daily subculturing into FAB (Fastidious Anaerobe Broth) fresh media. The study focuses on four individual phage-bacteria pairs: Bacteroides intestinalis APC919/174-crAss001, Bacteroides xylanisolvens APCS1/XY-crAss002, Bacteroides thetaiotaomicron VPI-5482 tdk-acapsular mutant (cps) - DAC15, and Parabacteroides distasonis APCS2/PD-PDS1. Each phage-host pair used had 4 biological replicates. The samples for the analysis were collected during the bacterial mid-log phase (OD600= ~0.3) on the fifth day of co-culture, which occurred between 5-7 hours after subculture. Bacterial cultures were centrifuged at 4,500 x g for 15 min at 4 °C. Immediately after centrifugation, the supernatant was transfer to a new tube and stored at -80 °C and the pellet was carefully dried to remove any remaining liquid and also stored at -80 °C. All samples were processed for proteomics analysis in which proteins differentially expressed in control or infected sample conditions were of interest due to their potential involvement during crAss-like phage infection.
Project description:Strain Bacteroides intestinalis TSDC17.2-1.1 (species Bacteroides intestinalis) was isolated from the fecal microbiota of a USA female at time point zero (bacterial isolates were sequenced from this donor on day 0 and 49). The species name was assigned by genome clustering.
Project description:Strain Bacteroides intestinalis TSDC20.2-1.2 (species Bacteroides intestinalis) was isolated from the fecal microbiota of a USA female at time point zero (bacterial isolates were sequenced from this donor on day 0 and 57). The species name was assigned by genome clustering.
Project description:Strain Bacteroides intestinalis TSDC20.2-2.1 (species Bacteroides intestinalis) was isolated from the fecal microbiota of a USA female at time point zero (bacterial isolates were sequenced from this donor on day 0 and 57). The species name was assigned by genome clustering.