Relationship between metabolism and biomass of medically important zygomycetes

Little is known about the relationships between metabolic activity and fungal biomass or time of incubation for medically important fungal pathogens. Understanding these relationships may be especially relevant for rapidly growing organisms, such as zygomycetes. A range of inocula of five clinical isolates of zygomycetes (one each of Rhizopus oryzae,Rhizopus microsporus, Cunninghamella bertholletiae, Mucor circinelloides and Absidia corymbifera) were incubated for 6, 8, 12, 24 and 48 h, after which hyphal mass was assessed spectrophotometrically and metabolic activity was measured using various concentrations of XTT and menadione. Both linear regression and the Boltzmann sigmoid model were used and compared for description of relationships between metabolic activity, biomass and time of incubation. Modeling was further applied to eleven additional zygomycete isolates. The relationships of biomass or metabolic activity as a function of time of incubation were well described with the Boltzmann sigmoid model. The latter was superior to linear regression in describing the relationship between metabolic activity and fungal biomass. For all isolates of zygomycetes, increases in metabolic activity preceded increases in biomass. Inter-species differences in growth patterns were observed, with Rhizopus microsporus and Mucor spp. reaching the plateau of growth earlier compared to other species. These findings on the temporal relationship and inter-species differences of hyphal growth and metabolic activity for zygomycetes may be useful in the design and interpretation of in vitro studies of these emerging pathogens.