The quantity of biomass is an essential parameter in growth and kinetic studies and also to characterize the optimum growth and sporulation conditions for different fungi during solid-state fermentation. In the case of solid-state fermentation, direct measurement of fungal biomass is hindered because fungi penetrate into and bind themselves tightly to the solid-substrate particles. Many researchers have described indirect methods to estimate biomass in solid state fermentation based on measuring the content of certain cell components like chitin, ergosterol and protein. The content of the different cell components can change markedly in fungi depending on fungal species, growth conditions and culture age.
Where as in the case of direct biomass estimation, duplicate samples of l g fermented substrate from each reactor has to transfer into to pre-weighed centrifuge tubes and 4 to 5 ml of sodium sulphate has to add in each tube and go for centrifugation at 12000 rpm for 20 minutes for a thrice under same conditions to achieve complete separation of biomass from the substrate. At the end of this step, the fungal mass with lower density than the substrate floated while the substrate settled to the bottom. The biomass alone has to transfer to a pre weighed filter paper and dried in hot air oven for 3 days to obtain a constant weight. Similarly, the substrate pellets in centrifuge tubes were also dried for 3 days and the record the weights. The remaining fermented substrate has to dry for 4 days to arrest further growth of the fungi and if necessary go for determination of biomass using indirect methods.
Thanks Chandra shekhar for a good answer. well this is a good method but not accurate since there might be some of the hyphae of fungus are also come with the fermented substrate, so it would be necessary to use indirect method could you suggest me the best and simple one.
I have the same problem and I didn't resolve it. But I have the idea that can be useful for you: plant reseachers use Phytagel - it is a substance that can substitute agar and can be removed after cultivation. For example, in Sigma:
http://www.sigmaaldrich.com/catalog/product/sigma/p8169?lang=en®ion=RU
In our studies, the fungal biomass is determined by measuring total nitrogen according to the Kjeldahl method with Nessler reagent after preboilin the samples in 0.5% of trichloroacetic acid for 15 min to remove non-protein components. True protein content is calculated as total nitrogen multiplied by 4.38.
Elisashvili sir, the method you suggested seems do-able. kindly provide a paper or a link for that protocol.
You can find my article "Evaluation and regulation of the lignocellulolytic activity of
novel white-rot basidiomycetes" in the Research Gate.
Since you remove the non-protein components, it looks like a valid method.
Nice suggestion. Either of direct or indirect method can be used, but in both the cases the results obtained will not be accurate.
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