The development of filamentous fungi in submerged culture

During the 24 hour period following inoculation, aggregation of spores and sporelings can have an important effect on the subsequent growth of filamentous fungi in submerged culture. This early phase of growth does not appear to have received much attention, and it was for this reason that the author's research was started. The aggregation, germination and early growth of the filamentous fungus Aspergillus niger have been followed in aerated tower fermenters, by microscopic examination. By studying many individual sporelings it has been possible to estimate the specific growth rate and germination times, and then to assess the branching characteristics of the fungus over a period of from 1 to 10 hours after germination. The results have been incorporated into computer models to simulate the development of the physical structure of individual and aggregated sporelings. Following germination, and an initial rapid growth phase, fungi were found to grow exponentially: in the case of A.niger the mean germination time was about 5 hours and the doubling time was as short as 1.5 hours. Branching also followed an exponential pattern and appeared to be related to hyphal length. Using a simple hypothesis for growth along with empirical parameters, typical fungal structures were generated using the computer models : these compared well with actual sporelings observed under the microscope. Preliminary work suggested that the techniques used in this research could be successfully applied to a range of filamentous fungi.

Clogging in horizontal subsurface flow constructed wetlands

Horizontal Subsurface Flow Treatment Wetlands (HSSF TWs) are used by Severn Trent Water as a low-cost tertiary wastewater treatment for rural locations. Experience has shown that clogging is a major operational problem that reduces HSSF TW lifetime. Clogging is caused by an accumulation of secondary wastewater solids from upstream processes and decomposing leaf litter. Clogging occurs as a sludge layer where wastewater is loaded on the surface of the bed at the inlet. Severn Trent systems receive relatively high hydraulic loading rates, which causes overland flow and reduces the ability to mineralise surface sludge accumulations. A novel apparatus and method, the Aston Permeameter, was created to measure hydraulic conductivity in situ. Accuracy is ±30 %, which was considered adequate given that conductivity in clogged systems varies by several orders of magnitude. The Aston Permeameter was used to perform 20 separate tests on 13 different HSSF TWs in the UK and the US. The minimum conductivity measured was 0.03 m/d at Fenny Compton (compared with 5,000 m/d clean conductivity), which was caused by an accumulation of construction fines in one part of the bed. Most systems displayed a 2 to 3 order of magnitude variation in conductivity in each dimension. Statistically significant transverse variations in conductivity were found in 70% of the systems. Clogging at the inlet and outlet was generally highest where flow enters the influent distribution and exits the effluent collection system, respectively. Surface conductivity was lower in systems with dense vegetation because plant canopies reduce surface evapotranspiration and decelerate sludge mineralisation. An equation was derived to describe how the water table profile is influenced by overland flow, spatial variations in conductivity and clogging. The equation is calibrated using a single parameter, the Clog Factor (CF), which represents the equivalent loss of porosity that would reproduce measured conductivity according to the Kozeny-Carman Equation. The CF varies from 0 for ideal conditions to 1 for completely clogged conditions. Minimum CF was 0.54 for a system that had recently been refurbished, which represents the deviation from ideal conditions due to characteristics of non-ideal media such as particle size distribution and morphology. Maximum CF was 0.90 for a 15 year old system that exhibited sludge accumulation and overland flow across the majority of the bed. A Finite Element Model of a 15 m long HSSF TW was used to indicate how hydraulics and hydrodynamics vary as CF increases. It was found that as CF increases from 0.55 to 0.65 the subsurface wetted area increases, which causes mean hydraulic residence time to increase from 0.16 days to 0.18 days. As CF increases from 0.65 to 0.90, the extent of overland flow increases from 1.8 m to 13.1 m, which reduces hydraulic efficiency from 37 % to 12 % and reduces mean residence time to 0.08 days.