I would like help with reproducibility in biofilm quantity and less time consuming (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species).
Dear Karthikeyan the most reproducible antibiofilm assay is the MBEC Assay System produced by innovatech. With best regards
http://www.innovotech.ca/products_biofilm.php
http://www.innovotech.ca/documents/MBECHTPInstructions_Rev2.pdf
Dear Bueno,
I am looking for lab study without antibiotic substance for the new antibiofilm agents devlopment....
choose the model that best suits your system. There are many different systems. find the best system that represents the fluid dynamics of your model. Substrate is also an important consideration since this will affect the binding of the biofilm and interaction with the inhibitory substance. Lastly the measurement of biofilm is important . are you measuring detachment or viability etc . Recently I have been looking into which was the best system as opposed to which was the most commonly used system. For rapid screens I use the crystal violet adhesion assay , 96 well high bind plates, (20C), can be agitated if you want but not higher than 10 rpm. Depending on the speed of growth of your test organism I do not use these past 2-3days. Also I limit the nutrients to stop the organisms growing too fast. the good thing about the system is that it is very easy to use but you have to do a lot of replicates. However if it's just a screen it is fast and easy. Typically i use a 1/100 dilution of an overnight culture to achieve the best biofilms. There are other systems like the calgary biofilm device and MBEC mentioned above which are more standardised. However you have to decide if you want to measure biofilm detachment or viability . Remember that some cells in a biofilm might not grow directly onto agar again but they may still be viable. I want to find a system that measures biofilm directly , ie cells + EPS . I have heard there are some good electric measurement systems but i am concerned about the presence of any electric contacts near the biofilms.
hope this is helpful
Have alook at www.biofilmcontrol.com, witi he biofilm ring test (r).
Contact me directly if you are interested in a partneship on anti-biofilm agent discovery.
Pascal
Crystal violet assay is good but you need to select your wells/plates to avoid variable evaporation.
This could be achieved by
1- fill all wells with 200ul poor medium of choice while knowing the exact weight in mg.
2- incubate plates in a humidified incubator.
3- place your plates on a specific corner in the incubator,
4- Measure remaining volume every 12 hrs using sensitive balance.
5- After 3 days, u'll be able to tell which wells consistently retained the maximum volume of liquid.
This way u'll suffer less from multiple replicates ..
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