I wish to determine biomass concentration from attached growth culture, grown on numerous hollow cylindrical cubes of volume 1 cc under anaerobic environment.
1. Has there been any latest modification to the methods described by Herbert et al. (1971) ("Chemical Analysis of Microbial Cells") and Liu et al. (1994) ("Simple Technique for Estimation of Biofilm Accumulation")?
2. If the aforesaid methods are still regarded as the best techniques to estimate attached biomass, which one, among the two, would be more effective towards determination of anaerobic attached biomass concentration and why?
Thanks in advance!
Hi Biswabandhu Chatterjee ,
are you growing the anaerobic biofilm in a defined medium free of suspended solids?
@Prof. Bachmann
Sir, the biofilm are basically attached onto mediums (small cylindrical cubes), which share the space in the anaerobic bioreactor with suspended biomass too.
I have been thinking to follow the modified Lowry method (1951).
I was thinking to suggest a gravimetric method but since the reactor contains suspended biomass (e.g. cellulose fibres) we would have to remove the suspended biomass from the biofilm (if it got attached/incorporated into the biofilm matrix). If the suspended biomass is not interfering you can try Article Thermogravimetric determination of biomass on activated carbon Environ
If the suspended biomass does not contain proteins, your idea should also work (e.g. Bradford, Lowry, TKN). You just need to know (from literature or your own study) the percentage of protein in the biofilm in order to convert the protein mass into biofilm biomass.
Another approach would be to determine the DNA content. A positive correlation between DNA and bacterial concentration was reported here: Article Fluorometric determination of the DNA concentration in munic...
You could also try chemical oxygen demand (COD) method if support material cannot be oxidised by COD reagent. Digest x numbers of hollow cylindrical support material with biofilm using closed reflux method (APHA 5220B; https://www.standardmethods.org/doi/abs/10.2105/SMWW.2882.103).
The following paper reviews Article Chemical and physical methods for characterisation of biofilms
which may be of general interest.Good luck and keep us posted on how you managed to determine biofilm biomass.
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