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Capillary bacterial migration on non-nutritive solid surfaces

Tomislav Ivanković
Tomislav Ivanković
, 
Uzi Hadad
Uzi Hadad
, 
Ariel Kushmaro
Ariel Kushmaro
, 
Svjetlana Dekić
Svjetlana Dekić
, 
Josipa Ćevid
Josipa Ćevid
, 
Marko Percela
Marko Percela
 et 
Jasna Hrenović
Jasna Hrenović
   | 06 oct. 2020
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Article Category: Original article
Publié en ligne: 06 oct. 2020
Pages: 251 - 260
Reçu: 01 mai 2020
Accepté: 01 août 2020
DOI: https://doi.org/10.2478/aiht-2020-71-3436
Mots clés
© 2020 Tomislav Ivanković et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Figure 1

Experimental setup for growing bacterial biofilms on an air/liquid interface
Experimental setup for growing bacterial biofilms on an air/liquid interface

Figure 2

Biofilms grown on glass microscopy slides at the air/liquid interface after 7 days of incubation. Far right: macroscopic view of biofilm formed at air/liquid interface. NS – experiments without shaking; S – experiments with shaking. Scale bar=50 μm
Biofilms grown on glass microscopy slides at the air/liquid interface after 7 days of incubation. Far right: macroscopic view of biofilm formed at air/liquid interface. NS – experiments without shaking; S – experiments with shaking. Scale bar=50 μm

Figure 3

Biofilm of P. aeruginosa at the air/liquid interface after 7 days of incubation under confocal microscopy. Left: autoflorescence; Right: bright field. Scale bar=50 μm
Biofilm of P. aeruginosa at the air/liquid interface after 7 days of incubation under confocal microscopy. Left: autoflorescence; Right: bright field. Scale bar=50 μm

Figure 4

Migration of B. cereus (A), A. junii (B), and S. aureus cells (C) across the air-exposed section of the glass slide imaged after 7 days of incubation. Measures in mm designate the distance crossed up the slide from the air/liquid interface. The “topmost visible microcolony” shows a microcolony or aggregate of cells reaching the farthest away from the interface
Migration of B. cereus (A), A. junii (B), and S. aureus cells (C) across the air-exposed section of the glass slide imaged after 7 days of incubation. Measures in mm designate the distance crossed up the slide from the air/liquid interface. The “topmost visible microcolony” shows a microcolony or aggregate of cells reaching the farthest away from the interface

Figure 5

Migration of A. baumannii cells across the air zone of glass microscopy slide imaged after 7 days of incubation. Measures in mm designate the distance crossed up the slide from the air/liquid interface. Upper row shows the “topmost visible microcolony”, a microcolony or aggregate of cells reaching the farthest away from the interface. This figure shows several snapshots from the same area (height)
Migration of A. baumannii cells across the air zone of glass microscopy slide imaged after 7 days of incubation. Measures in mm designate the distance crossed up the slide from the air/liquid interface. Upper row shows the “topmost visible microcolony”, a microcolony or aggregate of cells reaching the farthest away from the interface. This figure shows several snapshots from the same area (height)

Figure 6

Migration of P. aeruginosa cells across the air-exposed section of the glass slide at 20 mm above the air/liquid interface. Image on the right shows superimposed autofluorescence. Scale bar=50 μm
Migration of P. aeruginosa cells across the air-exposed section of the glass slide at 20 mm above the air/liquid interface. Image on the right shows superimposed autofluorescence. Scale bar=50 μm

Figure 7

Microcolonies of B. cereus (A and B) and B. thuringiensis (C) on the air-exposed section of the glass slide after 7 days of incubation. Images on the right show magnified cut-outs. EPS – extracellular polymeric substances. Scale bar=25 μm
Microcolonies of B. cereus (A and B) and B. thuringiensis (C) on the air-exposed section of the glass slide after 7 days of incubation. Images on the right show magnified cut-outs. EPS – extracellular polymeric substances. Scale bar=25 μm

Figure 8

Typical microcolony of B. cereus spotted high in the air-exposed section of the glass slide. Scale bar=25 μm
Typical microcolony of B. cereus spotted high in the air-exposed section of the glass slide. Scale bar=25 μm

Figure 9

Migration of A. junii (A) and A. baumannii (B) from the air/liquid interface to the air-exposed section of the glass slide. EPS is dyed in blue/purple to better show spreading up the slide [A. junii (C); A. baumannii (D)]
Migration of A. junii (A) and A. baumannii (B) from the air/liquid interface to the air-exposed section of the glass slide. EPS is dyed in blue/purple to better show spreading up the slide [A. junii (C); A. baumannii (D)]

Figure 10

Migration of carbol-fuchsin (left) and A. baumannii cells from nutrient media or saline (right) up the glass slide, recreated from experiments listed in Table 1. Starting bacterial concentrations were either 103 or 107 CFU/mL. The dashed line marks the farthest point reached on the given days of incubation
Migration of carbol-fuchsin (left) and A. baumannii cells from nutrient media or saline (right) up the glass slide, recreated from experiments listed in Table 1. Starting bacterial concentrations were either 103 or 107 CFU/mL. The dashed line marks the farthest point reached on the given days of incubation

Figure 11

Migration of carbol-fuchsin (left) and B. cereus cells from nutrient media or saline (right) up the glass slide, recreated from experiments listed in Table 1. Starting bacterial concentrations were either 103 or 107 CFU/mL. The dashed line marks the farthest point reached on the given days of incubation
Migration of carbol-fuchsin (left) and B. cereus cells from nutrient media or saline (right) up the glass slide, recreated from experiments listed in Table 1. Starting bacterial concentrations were either 103 or 107 CFU/mL. The dashed line marks the farthest point reached on the given days of incubation

Swarming and twitching motility determined by standard assays for B. cereus, A. junii, A. baumannii and P. aeruginosa and expressed as diameter of the growth area from inoculation spot

B. cereus A. junii A. baumannii P. aeruginosa
Swarming 82±14 mm N/A N/A 16±7 mm
Twitching N/A 35±12 mm 45±14 mm 51±9 mm

Experimental combinations performed in order to explain the migration of bacterial cells up the glass slide

Biofilm growth (days) B. cereus A. baumannii
103 CFU/mL 107 CFU/mL 103 CFU/mL 107 CFU/mL Carbol-fuchsin
1 Exp 1 Exp 2 Exp 17 Exp 18 Exp 33
LB medium 3 Exp 3 Exp 4 Exp 19 Exp 20 Exp 34
7 Exp 5 Exp 6 Exp 21 Exp 22 Exp 35
10 Exp 7 Exp 8 Exp 23 Exp 24 Exp 36
1 Exp 9 Exp 10 Exp 25 Exp 26 Exp 37
3 Exp 11 Exp 12 Exp 27 Exp 28 Exp 38
Sterile saline (0.3 %) 7 Exp 13 Exp 14 Exp 29 Exp 30 Exp 39
10 Exp 15 Exp 16 Exp 31 Exp 32 Exp 40
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