Packer, Helen Lyn; (1991) The Use of Image Analysis for the Characterisation of Filamentous Microorganisms. Doctoral thesis (Ph.D), UCL (University College London).

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Abstract

Characterisation of mycelial morphology is important for physiological and engineering studies of filamentous fermentations, and in the design and operation of such fermentations. Image analysis has been developed as a method for this characterisation and has been shown to be faster and generally more accurate than previous methods. A fully automatic system has been developed, in which speed has been gained, but with loss of accuracy in some cases. The method has been tested on Streptomyces clavuligerus and two strains of Penicillium chrysogenum, one of which was in a medium containing undissolved solids. Additionally a method for the estimation of biomass for the penicillin fermentation using image analysis is presented. Two regions of hyphae are defined to describe the growth of mycelia during fermentation 1) the cytoplasmic region and 2) the degenerated region including large vacuoles. The volume occupied by each of these regions in a fixed volume of sample is estimated from area measurements using image analysis. Areas are converted to volumes by treating the hyphae as solid cylinders with the hyphal diameter as the cylinder diameter. The volumes of the cytoplasmic and degenerated regions are converted into dry weight estimations using hyphal density values available in the literature. The image analysis method for biomass and cell volume estimation is shown to estimate successfully concentrations of mycelia from 0.03 to 38g/L for a number of fermentations of Penicillium chrysogenum. The image analysis method has also estimated cell volume and biomass even in the presence of 30g/L of undissolved solids in the medium. The image analysis estimation of biomass is shown to agree with the conventional method of measurement using filtration. Both image analysis methodologies are powerful tools for the quantification and characterisation of filamentous microorganisms and will allow greater understanding of engineering and physiological processes occurring during fermentation.

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