Aims: In solid state fermentation (SSF), estimation of biomass is difficult as fungal mycelium penetrates deep and remains attached to the solid substrate particles. This study examines and evaluates a new technique based on colour changes of fermented substrates during SSF as an indicator for fungal growth. Methodology and Results: SSF refers to microbial fermentation, which takes place in the absence or near absence of free water, thus being close to the natural environment in which the selected microorganisms, especially fungi, are naturally adapted. Although many promising methods are available, the evaluation of microbial growth in SSF may sometimes become difficult, impractical, and inaccurate. Essentially, this remains another critical issue for monitoring growth. In this study, measurements of colour changes of fermented substrates during SSF are used as indicators for growth and this technique has a potential to be used to quantify growth of microbes. For the growth of Aspergillus awamori and A. oryzae on wheat bran, soybean hulls, and rapeseed meal, it was confirmed that colour changes were directly proportional to the fungal growth. This new approach is an important complementation to the existing techniques, especially for basic studies. The advantages of this method are its ease of use, fast, non-destructive, cheap, and requires no special and expensive reagents. The key finding is that the colorimetric technique demonstrated in this study provides good means to estimate growth than that obtained by visual observation or spores counting.
Biomass ; Fermentation

Carleysmith and Fox (1984) stated “without doubt, the single most vital yet most problematical value sought duringfermentation is biomass estimation”. Achieving a positive result in determining biomass remains a major challenge insolid state fermentation (SSF). Fungi are well-characterised microorganisms and are widely used in SSF due to theirability to colonise and penetrate into the solid substrate. The compressed structure of the mycelia and the solid substratedoes not allow a complete recovery of the biomass, which may not be insurmountable. Since the use of a directtechnique such as the dry weight method is impractical, the use of an indirect estimation technique is the only alternative.This review examines strategies that have been used to estimate biomass in SSF. Many promising indirect estimationtechniques are available, which can be classified into six categories as follows; (i) measuring cell components notpresent in the substrate; (ii) measuring biomass component present in both substrate and biomass; (iii) measuring othersecondary metabolites; (iv) measuring metabolic activity; (v) measuring images from direct microscopic observation and(vi) measuring biomass from the substrate matrix. New potential technique and future directions are also discussed inthis review. Although significant advances have been made with the availability of various techniques; however, progresshas been very unsatisfactory. The evaluation of microbial growth in SSF may sometimes become laborious, impracticaland inaccurate. Essentially, this remains another critical issue for monitoring growth. The information of the profile offungal biomass growth throughout any SSF process constitutes an essential parameter in estimation of kinetic variablesand subsequently, scale-up of the process.

Aims:In this study, measurement of colour changes during solid state fermentation (SSF) are presented as one of the potential techniques that can be used to describe growth, complementary to other biomass estimation, such as weight of fungus, spores concentration, organic matter loss, glucosamine and enzyme activity, which is directly related to growth.Methodology and results: In this study, fungal fermentation of Aspergillus awamoriand A.oryzaeare carried out on complex heterogeneous solid media; wheat bran, soybean hull and rapeseed meal, which are constituted of various soluble and insoluble solid particles. Fermented mass was extracted using distilled water to obtain a cell free extract, which canbe determined quickly and accurately using UV-Visible spectrophotometermeasured absorbance at 300 nm. The results showed a significant correlation between colours produced from fungal SSF and the concentration of spores, weight of fungus cells, organic matter loss, glucosamine and enzyme activity. We found that the colour density proportionally increased when the studied parameters mentioned above increased.For the growth of A. awamoriand A. oryzaeon wheat bran, soybean hulls and rapeseed meal, it was confirmed that colour production was directly proportional to fungal growth. In general, colour-based methods seem to be the most promising approach for biomass estimation in SSF.Conclusions, significance and impacts of study:The theory of measuring colour changes in SSF by UV-visible spectroscopy demonstrates that the colour method gives some valuableinformation than just obtaining a visual observation or spore count to describe growth of fungal mycelium in SSF. The ideas obtained from this exercise might provide a quick and convenient method for quality control of fungal growth. The advantages of this method are that the procedure is simple to carry out, it is non-destructive and no special and expensive reagents are required and the process is very cheap.This newapproach is an important complimentation to the existing techniques especially for basic studies.