Be careful, it is dangerous.

dear Omar

I will send in the attach file M.Sc thesis which was worked under my supervision which correlates with technique in lab biofilm formation. Hope to be interested. Best regard

Use swab for collection of samples. The swab test method has proved a popular testing method, where there may be sewage contamination. If swab samples are collected for culture analysis, they should be sent to the laboratory within 24 hours after collection. If the analysis of the swab samples involves enumeration of the microbial contaminants, the size of the area sampled should be provided to the lab. Use specific media for ciulturing Pseudomonas.

Thanks dr Mushtak,Dr Pramod for answers .Thanks dr hussain for precaution .

The crystal violet 96-well plate assay is the easiest. However, this can be difficult when working with Pseudomonas as it forms a thick slime which can effectively pull the biofilm off the well when aspirating steps are performed. What I have found is if you aspirate using a pipette and are very careful you can remove the thicken media and leave the adhered biofilm on the well plate edges and bottom.

Alternatively, one can grow biofilm on a plastic or glass slide which is placed into a liquid culture of the bacteria (use whichever material you find is best for the bacteria to form biofilms on). Use CV to quantify or other methods such as live dead staining (microscopy can be performed easily with this method). Another advantage to this is the ability to change media and grow biofilms over several cycles of fresh media making the biofilm thicker each cycle. The downside is it is much less high throughput than the 96-well plate assay.

If you want to get quantitative results you can use the method of cristal violet, as Donald recomended. As he said, you need to be carefull at the steps in wich you remove the liquids to avoid dragging of the biofilm. Another method that you can use is congo red. This method is not quantitative (as far as I know) but is easy to perform. You just grow your strains in plates containing congo red and access the colony morphology (wrinkle or smooth) and congo red incorporation into the colony. Both things, rugosity and colour gives you information about biofilm formation capability of the strain.

Take in account that you always need a referecnce strain, I mean all this techniques are comparative, you need something to compare with and relativize your results.

Hi Balram,

You can try this method:

Article Fluorescence-based real-time monitoring of Pseudomonas aerug...

Good luck,