Hello, I am new to biofilm research and have a question about growing and measuring biofilms.
We want to research the effect of a type of enzymes in breaking down biofilms in showers, baths, hot tubs, and hot springs.
Therefore we must devise a way to grow and measure biofilm that is close to natural biofilms in showers etc.
Are there any existing protocols or any other researches that may be useful?
Dear Nanami,
This is a very interesting question - even though I am not working in this area - because it is not always the case that you get to design your own assay to investigate something.
Sometime ago we were interested to see if our particular biofilm-forming mutant, known as a Wrinkly Spreader and derived from Pseudomonas fluorescens SBW25, could 'evolve' in conditions very different to their normal environment (static liquid broth cultures). We eventually developed a glass-bead column system which was feed by a very slow peristaltic pump dripping media onto a syringe column packed with small glass balls. We connected this to an out-flow tube to make sure that the glass balls were never flooded (i.e producing a partially-saturated pore network between the glass balls with a set matrix pressure), providing bacteria with a shallow liquid layer that was constantly flowing away from the biofilm-colonisation site.
Perhaps you could design a similar system where patches of shower curtain material are being constantly washed by fresh media - you would need to do this in replicate, of course, but perhaps you could cut out patches all of the same size, and curl them into 20 ml plastic syringes and drip media onto them using a multi-tube pump system. If you could autoclave the syringe plus curtain 'microcosms' you could prepare many in advance and use them when needed.
You could develop this type of approach in lots of different ways, and cold even put in small sections of wood, plastic, stone or metal surfaces if you wanted. However, we tried growing biofilms on different types of stone, and some times it is very difficult to recover bacteria from some surfaces, or to stain them for quantitative measurement - so it is important to make sure that you have got a good negative control and assay protocol tested before you start your main experiment!
Regards, Andrew.
Dear Nanami, the following papers maybe helpfull to find answer to your question.
You can isolate bacteria from surfaces and than you can characterize their biofilm formation capabilities on different materials surfaces like stainless steel or polymer surfaces. There are both quantitative and qualitative methods to evaluate biofilm formation.
Article Identification and characterization of microbial biofilm com...
- https://doi.org/10.1007/s11665-014-1332-x Effect of Mixed-Species Biofilm on Copper Surfaces in Cooling Water System
There are protocolos and advice. But maybe it depend on how you set the experiment
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