Template:Team:Groningen/CONTENT/LOGBOOK/Grow a Biofilm on a Carrier Material
Grow a Biofilm on a Carrier Material
Grew B. subtilis 3610 ComI on different carrier materials and with different media to find ideal conditions for biofilm growth.
Find the best carrier material and medium for the biofilm of B. subtilis 3610 ComI.
Biofilm growth thrives best in a minimal medium like MSgg. LB medium is too rich with nutrients for the bacteria to trigger solid biofilm growth The biofilms seen on LB are not more than a smear of bacteria. On MSgg medium, biofilms form interesting structures.
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00:00, 25 March 2015 - 00:00, 26 March 2015
Plate 10 µL B. subtilis 3610 ComI overnight culture (OD: 2,28 (600nm) after 22h growth) on different carrier materials on LB agar and MSgg agar plates. MSgg was prepared according to protocol.
Carrier materials using LB agar as a medium
Nylon positive charge (Boeringer, 1269299)
Nytran Plus 0.45 µm (Schleicher en Schuell, AH0495-1)
PVDF (Rocke, 03010040001)
Nitrocellulose 0.2 µm (invitrogen, LC2000)
Nylon 0.45 µm (Merck Milipore, HNWP04700)
Nitrocellulose 0.025 µm (Merck Milipore, VSWP02500)
Control (no carrier)
Carrier materials using MSgg agar as a medium
Nytran Plus 0.45 µm (Schleicher en Schuell, AH0495-1)
PVDF (Rocke, 03010040001)
Nitrocellulose 0.2µm (invitrogen, LC2000)
Nylon 0.45 µm (Merck Milipore, HNWP04700)
Nitrocellulose 0.025µm ( Merck Milipore, VSWP02500)
Control (no carrier)
All plates with carrier and bacteria were put in the incubator at 30°C.
00:00, 26 March 2015 - 00:00, 26 March 2015
After 24 hours, the biofilm growth was checked.
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LB plates after 24 hours. A is the control, B: Nitrocellulose 0.025µM, C: Nitrocellulose 0.2µM, D: Nylon 0.45µM, E: Nytran Plus 0.45µM, F: Nylon positive charge and G: PVDF. On all but the PVDF membrane are colonies growing.
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MSgg plates after 24 hours. A: control, B: Nitrocellulose 0.2µM, C: Nitrocellulose 0.025µM, D: Nytran Plus 0.45µM, E: PVDF and F: Nylon 0.45µM. Bacterial growth is seen on all samples, except for PVDF.
00:00, 30 March 2015 - 00:00, 30 March 2015
After 120 hours, the biofilm growth was checked. Biofilm growth thrives better on MSgg as can be seen on the images below. After 120 hours there is a clear difference visible between the LB and MSgg conditions. Furthermore, Nitrocellulose 0.2µM shows an nice and even distribution of biofilm and PVDF doesn't show biofilm growth at all on the MSgg plate.
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LB plates after 120 hours. A is the control, B: Nitrocellulose 0.025µM, C: Nitrocellulose 0.2µM, D: Nylon 0.45µM, E: Nytran Plus 0.45µM, F: Nylon positive charge and G: PVDF. Bacterial biofilm growth is seen on all the carrier materials.
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MSgg plates after 24 hours. A: control, B: Nitrocellulose 0.2µM, C: Nitrocellulose 0.025µM, D: Nytran Plus 0.45µM, E: PVDF and F: Nylon 0.45µM. Bacterial growth is seen on all samples, except for PVDF.