Journal: Viruses

Article Title: Phage-Mediated Digestive Decolonization in a Gut-On-A-Chip Model: A Tale of Gut-Specific Bacterial Prosperity

doi: 10.3390/v16071047

Figure Lengend Snippet: GOAC and OmniLog bacteria-only control conditions. Bacterial titers’ evolution in GOACs when infected with P. aeruginosa CN573 (10 7 CFU/m; n = 3) ( A ) or E. coli ST131 at either higher (10 7 CFU/mL; n = 3) ( B ) or lower (10 5 CFU/mL; n = 3) initial titer concentrations ( C ). Bacterial control growth curves in static in vitro conditions using OmniLog automated incubator, with bacterial respiration units being measured as proxy for bacterial growth (n = 16 wells for each condition; initial bacterial load of 10 5 CFU/well; error bars represent +/−1 standard deviation of mean) ( D ). Microscopic view of GOAC inlet catheter transversal histological section after 48 h P. aeruginosa CN573 10 7 CFU/mL colonization after Gram staining. P. aeruginosa appear as numerous typical Gram-negative pink-red bacili that seemingly form homogeneously adherent lining to apical pole of continuous Caco-2 GOAC epithelium (e), and can also be seen forming consortium in likely apical mucus lining (m, grey). ( E ) Microscopic view of GOAC inlet catheter transversal histological section after 48 h P. aeruginosa CN573 10 7 CFU/mL colonization after Gram staining. P. aeruginosa appear as numerous typical Gram-negative pink-red bacilli that seemingly form homogeneously adherent lining to apical pole at continuous Caco-2 GOAC epithelium ( e ) and can also be seen forming consortium in likely apical mucus lining ( m , grey). [Pa: P. aeruginosa; CFU: colony forming unit; L: luminal; P: parietal] .

Article Snippet: To compare GOACs’ bacterial dynamics to other in vitro conditions over an identical timeframe of 72 h, bacterial growth curves were established using an OmniLog incubator system (Biolog, Hayward, CA, USA).

Techniques: Bacteria, Control, Infection, In Vitro, Standard Deviation, Staining

Journal: Viruses

Article Title: Phage-Mediated Digestive Decolonization in a Gut-On-A-Chip Model: A Tale of Gut-Specific Bacterial Prosperity

doi: 10.3390/v16071047

Figure Lengend Snippet: P. aeruginosa CN573 versus phage PNM assays. P. aeruginosa titer evolution when introduced in GOACs with phage PNM (MOI = 1) in various conditions: ( A ) simultaneous introduction of P. aeruginosa and PNM in 100% Caco-2 GOACs (dotted line represents detection threshold in spread plating, <10 2 CFU/mL) (n = 3), ( B ) sequential introduction of P. aeruginosa first with subsequent introduction of phage PNM after 1 h static incubation delay in 100% Caco-2 GOACs (n = 3), ( C ) replication of these two conditions in bicellular GOACs (n = 3 each). ( D ) P. aeruginosa and PNM (MOI = 1) control growth curves in static in vitro conditions using OmniLog automated incubator, with bacterial respiration units being measured as proxy for bacterial growth (initial bacterial load of 10 5 CFU/well; n = 32 wells for P. aeruginosa + PNM, 16 wells for P. aeruginosa only, and 16 wells for negative control; error bars represent +/−1 standard deviation of mean). ( E ) Spread plating on 5% sheep blood Columbia agar plates highlights intense phenotypic diversification in P. aeruginosa CN573 after co-evolution with phage PNM in GOACs, displaying at least four distinct colony morphologies out of single GOAC after 48 h. ( F ) Close-up view of one of these plates reveals co-existence of wild-type colonies (white arrow), systematically including phage-induced lysis plaques, along with modified colonies, for example, exhibiting dark pigmented halo (black arrow), systematically devoid of any lysis plaques. [Pa: P. aeruginosa; GOAC: Gut-On-A-Chip; MOI: Multiplicity of Infection] .

Article Snippet: To compare GOACs’ bacterial dynamics to other in vitro conditions over an identical timeframe of 72 h, bacterial growth curves were established using an OmniLog incubator system (Biolog, Hayward, CA, USA).

Techniques: Incubation, Control, In Vitro, Negative Control, Standard Deviation, Lysis, Modification, Infection

Journal: Viruses

Article Title: Phage-Mediated Digestive Decolonization in a Gut-On-A-Chip Model: A Tale of Gut-Specific Bacterial Prosperity

doi: 10.3390/v16071047

Figure Lengend Snippet: E. coli ST131: dual phage and OmniLog assays. E. coli titer evolution when introduced in bicellular GOACs with 50%/50% combination of phage ES17 and phage HP3 in comparison with both mono-phage counterparts at MOI = 1 (n = 3 each) ( A ) or MOI = 10 (n = 3 each) ( B ). Example of modified E. coli ST131 colony morphology (left, variant ST131-V1 see ) after exposition to HP3 at MOI = 1 compared to wild-type colony morphology (right) ( C ). E. coli with either ES17 ( D ) or HP3 ( E ) control growth curves in static in vitro conditions using OmniLog automated incubator, with bacterial respiration units being measured as proxy for bacterial growth (initial bacterial load of 10 5 CFU/well; n = 24 wells for each MOI = 1 assay, 8 wells for each MOI = 10 assay, 16 wells for E. coli only, and 16 wells for negative control; error bars represent +/−1 standard deviation of mean). [Ec: E. coli; MOI: Multiplicity of Infection] .

Article Snippet: To compare GOACs’ bacterial dynamics to other in vitro conditions over an identical timeframe of 72 h, bacterial growth curves were established using an OmniLog incubator system (Biolog, Hayward, CA, USA).

Techniques: Comparison, Modification, Variant Assay, Control, In Vitro, Negative Control, Standard Deviation, Infection

Journal: Viruses

Article Title: Phage-Mediated Digestive Decolonization in a Gut-On-A-Chip Model: A Tale of Gut-Specific Bacterial Prosperity

doi: 10.3390/v16071047

Figure Lengend Snippet: Serum-supplemented OmniLog assays. Replicating previous OmniLog assays with the supplementation of 20% fetal bovine serum (the same proportion as in the GOAC culture medium) to LB yields significantly higher bacterial escape growth rates in P. aeruginosa –PNM assays ( A , B ), but not in E. coli –ES17 ( C , D ) nor in E. coli –HP3 ( E , F ) assays.

Article Snippet: To compare GOACs’ bacterial dynamics to other in vitro conditions over an identical timeframe of 72 h, bacterial growth curves were established using an OmniLog incubator system (Biolog, Hayward, CA, USA).

Techniques: