Difference between revisions of "Template:Team:Groningen/CONTENT/LOGBOOK/Grow a Biofilm on a Carrier Material"

Latest revision as of 09:41, 15 September 2015

Grow a Biofilm on a Carrier Material

Bacteria are triggered by their environment to change their behavior. Bacteria, like B. subtilis, preferably grow on rich media. However, when the medium where they grow on is less rich in nutrients, they start to form biofilms. In our project we need a carrier material to grow our biofilm on to make it more stable. In this experiment B. subtilis was grown on different media and different carrier materials.

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. Biofilms grown on MSgg show more structural integrity than biofilms grown on LB. Although the biofilm on the Nitrocellulose 0.025µM looks nice, the biofilms on the carrier materials with larger pores (0.2µM-0.45µM) are more suited for our project since these biofilms are interwoven with the carrier.

<a class="postscriptum protocol" href="https://2015.igem.org/Team:Groningen/Protocols_and_Protocols/No_protocol">Finibus sapien, nec cursus...</a>

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.

Harm Ruesink

00:00, 26 March 2015 - 00:00, 26 March 2015

After 24 hours, the biofilm growth was checked.

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.

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.

Harm Ruesink

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.

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.

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.

Harm Ruesink

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 	|goal=<html>Find the best carrier material and medium for the biofilm of B. subtilis 3610 ComI.
 </html>
-	|description=<html>Grow B. subtilis 3610 ComI on different carrier materials and with different media to find the ideal conditions for its biofilm growth.
+	|description=<html> Bacteria are triggered by their environment to change their behavior. Bacteria, like B. subtilis, preferably grow on rich media. However, when the medium where they grow on is less rich in nutrients, they start to form biofilms. In our project we need a carrier material to grow our biofilm on to make it more stable. In this experiment B. subtilis was grown on different media and different carrier materials.
 </html>
-	|conclusion=<html>Nulla pellentesque odio sit amet orci sollicitudin vulputate. Nulla aliquet ante a luctus efficitur. Nulla iaculis dolor eget diam tempor, in facilisis nisl placerat. Suspendisse sed felis metus. Sed nibh dolor, hendrerit at nibh vitae, posuere porttitor odio. Sed eget consequat urna. Maecenas a dolor ut massa pulvinar finibus non ac justo. Nam pulvinar, quam quis placerat tincidunt, tortor nunc sagittis orci, sed ullamcorper nulla nibh eu justo. Duis malesuada velit a metus malesuada eleifend. Ut at libero sem. Cras vel magna luctus, tristique mi sit amet, commodo odio. Vestibulum gravida lorem a mi auctor facilisis. Cras sodales aliquet massa sed vestibulum. Aliquam id ornare elit. Aenean vel justo ut nisi tincidunt aliquet et id ex. Mauris tristique velit mollis velit facilisis facilisis a vel nulla.
+	|conclusion=<html>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. Biofilms grown on MSgg show more structural integrity than biofilms grown on LB. Although the biofilm on the Nitrocellulose 0.025µM looks nice, the biofilms on the carrier materials with larger pores (0.2µM-0.45µM) are more suited for our project since these biofilms are interwoven with the carrier.
 </html>
 }}