Impact of a human gut microbe on Vibrio cholerae host colonization through biofilm enhancement
- Department of Molecular Biology and Microbiology, Tufts University School of Medicine, United States
- Program of Molecular Microbiology, Graduate School of Biomedical Sciences, Tufts University School of Medicine, United States
- Department of Medicine, University of Washington, United States
- Division of Infectious Diseases, Massachusetts General Hospital, United States
- Department of Molecular, Cellular and Developmental Biology, Yale University, United States
- Department of Pediatrics, Harvard Medical School, United States
- Department of Molecular Virology and Microbiology at Baylor College of Medicine, United States
- International Center for Diarrheal Disease Research, Bangladesh, Bangladesh
- Quantitative Biology Institute, Yale University, United States
Figures
Figure 1
Paracoccus aminovorans (Pa) is more abundant in persons with Vibrio cholerae (Vc) infection compared to uninfected persons.
In a prior study of household contacts of cholera patients in Bangladesh (Midani et al., 2018), Pa was identified as differentially abundant using a support vector machine model with recursive …
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Figure 1—source data 1
Paracoccus aminovorans (Pa) abundance in persons with and without Vibrio cholerae (Vc) infection.
- https://cdn.elifesciences.org/articles/73010/elife-73010-fig1-data1-v3.xlsx
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Figure 1—source data 2
Ratio of Paracoccus aminovorans (Pa) to Vibrio cholerae (Vc) in infected persons.
- https://cdn.elifesciences.org/articles/73010/elife-73010-fig1-data2-v3.xlsx
Figure 2 with 2 supplements
The presence of Paracoccus aminovorans (Pa) enhances Vibrio cholerae (Vc) colonization in the infant mouse intestine.
(A) 3-day-old infant mice were intragastrically inoculated with 107 colony-forming unit (CFU) of Pa every 12 hr twice or four times. 12 hr after the second or fourth dose of Pa inoculation (i.e., at …
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Figure 2—source data 1
Viable cell count of Paracoccus aminovorans (Pa) in infant mouse intestine.
- https://cdn.elifesciences.org/articles/73010/elife-73010-fig2-data1-v3.xlsx
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Figure 2—source data 2
Viable cell count of Vibrio cholerae (Vc) in the infant mouse intestine with and without Pa pre-colonization.
- https://cdn.elifesciences.org/articles/73010/elife-73010-fig2-data2-v3.xlsx
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Figure 2—source data 3
Viable cell count of Vibrio cholerae (Vc) in the infant mouse intestine with and without Pa co-infection.
- https://cdn.elifesciences.org/articles/73010/elife-73010-fig2-data3-v3.xlsx
Figure 2—figure supplement 1
Paracoccus aminovorans (Pa) colonization does not significantly alter the mouse gut microbial diversity.
3-day-old infant mice were intragastrically inoculated with 107 colony-forming unit (CFU) of Pa or sterile LB medium as control (Ctrl) for a period of 12 hr to mimic the length of time between the …
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Figure 2—figure supplement 1—source data 1
Abundance of microbes in the mouse intestine with and without Pa colonization.
- https://cdn.elifesciences.org/articles/73010/elife-73010-fig2-figsupp1-data1-v3.xlsx
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Figure 2—figure supplement 1—source data 2
Principal Component Analysis of microbial abundance in mouse intestine with and without Pa colonization.
- https://cdn.elifesciences.org/articles/73010/elife-73010-fig2-figsupp1-data2-v3.xlsx
Figure 2—figure supplement 2
Paracoccus aminovorans (Pa) colonization significantly increases Vibrio cholerae (Vc) small intestine colonization.
(A, B) 3-day-old infant mice were intragastrically inoculated with 107 colony-forming unit (CFU) of Pa or sterile LB medium as control for a period of 36 hr. These animals were then infected with Vc …
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Figure 2—figure supplement 2—source data 1
Viable cell count of Vibrio cholerae (Vc) in the mouse small intestine with and without Pa pre-colonization.
- https://cdn.elifesciences.org/articles/73010/elife-73010-fig2-figsupp2-data1-v3.xlsx
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Figure 2—figure supplement 2—source data 2
Viable cell count of Paracoccus aminovorans (Pa) and Vibrio cholerae (Vc) in the mouse small intestine 6 hours after Vc infection.
- https://cdn.elifesciences.org/articles/73010/elife-73010-fig2-figsupp2-data2-v3.xlsx
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Figure 2—figure supplement 2—source data 3
Viable cell count of Paracoccus aminovorans (Pa) in the mouse intestine with and without Vibrio cholerae (Vc) infection.
- https://cdn.elifesciences.org/articles/73010/elife-73010-fig2-figsupp2-data3-v3.xlsx
Figure 3 with 1 supplement
Paracoccus aminovorans (Pa) promotes biofilm formation of Vibrio cholerae (Vc).
(A) Planktonic cell counts from cultures used for pellicle analysis of Vc and Pa grown together or in monoculture. (B) Representative images of pellicles formed by Vc grown in monoculture and …
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Figure 3—source data 1
Viable cell count in for Paracoccus aminovorans (Pa) and Vibrio cholerae (Vc) in the planktonic phase in mono- and co-cultures.
- https://cdn.elifesciences.org/articles/73010/elife-73010-fig3-data1-v3.xlsx
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Figure 3—source data 2
Pellicle formation in Vc mono- and Vc/Pa coculture.
- https://cdn.elifesciences.org/articles/73010/elife-73010-fig3-data2-v3.pdf
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Figure 3—source data 3
Viable cell count in for Paracoccus aminovorans (Pa) and Vibrio cholerae (Vc) in the pellicle formed by mono- and co-cultures.
- https://cdn.elifesciences.org/articles/73010/elife-73010-fig3-data3-v3.xlsx
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Figure 3—source data 4
Viable cell count in for Paracoccus aminovorans (Pa) and Vibrio cholerae (Vc) released by washing of the pellicle formed by mono- and co-cultures.
- https://cdn.elifesciences.org/articles/73010/elife-73010-fig3-data4-v3.xlsx
Figure 3—figure supplement 1
Paracoccus aminovorans (Pa) cultures increase biofilm production in Vibrio cholerae (Vc).
(A) Biofilm formation assays were performed in 96-well microtiter plates. Pa was diluted in LB to a concentration of 107 colony-forming unit (CFU) and grown for 24 hr before the addition of WT Vc. …
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Figure 3—figure supplement 1—source data 1
Crystal violet assays of biofilm formed by Vc mono- and Vc/Pa co-cultures.
- https://cdn.elifesciences.org/articles/73010/elife-73010-fig3-figsupp1-data1-v3.xlsx
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Figure 3—figure supplement 1—source data 2
Crystal violet assays of biofilm formed by Vc mono- and Vc/Pa co-cultures using vpsL mutants with and without complementation plasmids.
- https://cdn.elifesciences.org/articles/73010/elife-73010-fig3-figsupp1-data2-v3.xlsx
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Figure 3—figure supplement 1—source data 3
Viable cell count of of pellicles formed by Vc mono- and Vc/Pa co-cultures using vpsL mutants with and without complementation plasmids.
- https://cdn.elifesciences.org/articles/73010/elife-73010-fig3-figsupp1-data3-v3.xlsx
Figure 4 with 1 supplement
Representative microscopy images of Vibrio cholerae (Vc) and Paracoccus aminovorans (Pa) dual-species pellicles.
(A) Large-scale cross-sectional image of the internal structure in a co-culture pellicle. All cells are stained with FM 4-64 and Vc cells constitutively express mNeonGreen. Therefore, the red signal …
Figure 4—figure supplement 1
Negative controls for wheat germ agglutinin (WGA) staining and the reporter strain.
(A) Representative view of Paracoccus aminovorans (Pa) cell monoculture imaged under identical conditions as in Figure 6A and B. Scale bar: 20 µm. (B) Zoom-in view of the region highlighted by the …
Figure 5
Paracoccus aminovorans (Pa) increases biofilm production in Vibrio cholerae (Vc).
Crystal violet assays were performed in 96-well microtiter plates to quantify biofilm formation. Overnight-grown (A) wild-type Vc or (B) ∆vpsL mutant and Pa cultures were diluted to a final …
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Figure 5—source data 1
Crystal violet assays of biofilm formed by Vc mono- and Vc/Pa co-cultures using WT with live or heat-killed Pa.
- https://cdn.elifesciences.org/articles/73010/elife-73010-fig5-data1-v3.xlsx
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Figure 5—source data 2
Crystal violet staining of biofilm formed by Vc mono- and Vc/Pa co-cultures using vpsL mutants with live and heat-killed Pa.
- https://cdn.elifesciences.org/articles/73010/elife-73010-fig5-data2-v3.xlsx
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Figure 5—source data 3
qRT-PCR analysis of vpsL gene in cells in biofilm formed by Vc mono- and Vc/Pa co-cultures.
- https://cdn.elifesciences.org/articles/73010/elife-73010-fig5-data3-v3.xlsx
Figure 6
Vibrio cholerae/Paracoccus aminovorans (Vc/Pa) co-culture biofilms depend on Vibrio exopolysaccharide (VPS).
(A) Representative cross-sectional view of the bottom layer of a co-culture pellicle with wheat germ agglutinin (WGA) staining. Vc cells constitutively express SCFP3A cytosolically; all cells were …
Figure 7 with 2 supplements
Enhanced Vibrio cholerae (Vc) intestinal colonization in the presence of Paracoccus aminovorans (Pa) is dependent on Vibrio exopolysaccharide (VPS) and accessory matrix proteins.
(A) 3-day-old infant mice were intragastrically inoculated with LB or 107 colony-forming unit (CFU) of Pa every 12 hr for a period of 48 hr, and subsequently infected with 106 CFU of a Vc strain …
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Figure 7—source data 1
Viable cell count of Vc in the mouse intestine with and without Pa pre-colonization using Vc WT and vpsL mutants.
- https://cdn.elifesciences.org/articles/73010/elife-73010-fig7-data1-v3.xlsx
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Figure 7—source data 2
Viable cell count of Vc in the mouse intestine with and without Pa pre-colonization using Vc WT and different biofilm matrix protein mutants.
- https://cdn.elifesciences.org/articles/73010/elife-73010-fig7-data2-v3.xlsx
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Figure 7—source data 3
Crystal violet staining of biofilm formed by different biofilm matrix protein mutants with and without Pa.
- https://cdn.elifesciences.org/articles/73010/elife-73010-fig7-data3-v3.xlsx
Figure 7—figure supplement 1
ctxA and tcpA gene expression in Vibrio cholerae (Vc) monoculture and Vibrio cholerae-Paracoccus aminovorans (Vc-Pa) co-culture.
Vc and Pa were co-cultured (1:10) statically for 24 hr, and bacteria in the air–liquid interface were sampled and RNA extracted. The relative ctxA and tcpA transcript levels were determined by …
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Figure 7—figure supplement 1—source data 1
qRT-PCR analysis of virulence genes in Vc with and without Pa.
- https://cdn.elifesciences.org/articles/73010/elife-73010-fig7-figsupp1-data1-v3.xlsx
Figure 7—figure supplement 2
Complementation of vpsL mutant in vivo.
The loss of colonization enhancement of ∆vpsL mutants in the presence of Paracoccus aminovorans (Pa) is restored by the introduction of a plasmid constitutively expressed vpsL. Mann–Whitney U-test, …
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Figure 7—figure supplement 2—source data 1
Viable cell count of Vc in mouse intestine with and without Pa pre-colonization using vpsL mutant with and without vpsL complementation plasmid.
- https://cdn.elifesciences.org/articles/73010/elife-73010-fig7-figsupp2-data1-v3.xlsx
Additional files
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Supplementary file 1
Raw and normalised counts of Pa and Vc.
(a) Operational taxonomic unit raw and normalized abundance of Vibrio cholerae (Vc) in Paracoccus aminovorans (Pa) colonized and Pa noncolonized study participants with Vc infection. (b) Raw and normalized abundance of total, Pa, and Vc operational taxonomic units in all study participants.
- https://cdn.elifesciences.org/articles/73010/elife-73010-supp1-v3.xlsx
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Transparent reporting form
- https://cdn.elifesciences.org/articles/73010/elife-73010-transrepform1-v3.pdf
