Wastewater treatment plants with enhanced biological phosphorus removal represent a state-of-the-art technology. Nevertheless, the process of phosphate removal is prone to occasional failure. One reason is the lack of knowledge about the structure and function of the bacterial communities involved. Most of the bacteria are still not cultivable, and their functions during the wastewater treatment process are therefore unknown or subject of speculation. Here, flow cytometry was used to identify bacteria capable of polyphosphate accumulation within highly diverse communities. A novel fluorescent staining technique for the quantitative detection of polyphosphate granules on the cellular level was developed. It uses the bright green fluorescence of the antibiotic tetracycline when it complexes the divalent cations acting as a countercharge in polyphosphate granules. The dynamics of cellular DNA contents and cell sizes as growth indicators were determined in parallel to detect the most active polyphosphate-accumulating individuals/subcommunities and to determine their phylogenetic affiliation upon cell sorting. Phylotypes known as polyphosphate-accumulating organisms, such as a "Candidatus Accumulibacter"-like phylotype, were found, as well as members of the genera Pseudomonas and Tetrasphaera. The new method allows fast and convenient monitoring of the growth and polyphosphate accumulation dynamics of not-yet-cultivated bacteria in wastewater bacterial communities.
Chitosan is a major structural component of fungal cell walls and has diverse medical and other applications. However, cost‐effective culture and extraction methods for fungi need to be developed. Therefore, Mucor rouxii was grown on YPG‐media in both submerged batch and semi‐continuous cultures. Chitosan was extracted from the mycelia to explore strategies to enhance yields and production rates. As observed in earlier studies, M. rouxii is able to adapt to shear stress when cultured semi‐continuously. Modeling the hyphal growth of batch experiments shows that the mycelia were ruptured by shear forces within a short cultivation time shown by a decreased hyphal length. However, an increasing chitosan content was observed with an increasing cultivation period in semi‐continuous cultures, which is an indication for the adaption to shear stress. Semi‐continuous culture resulted in the highest contents of extractable chitosan. The results and models of hyphal growth, including tip extension and branching, suggest that repeated batch cultures may be optimal for chitosan production.
Downstream processing of chitosan requires several technological steps that contribute to the total production costs. Precipitation and especially evaporation are energy-consuming processes, resulting in higher costs and limiting industrial scale production. This study investigated the filtration kinetics of chitosan derived from cell walls of fungi and from exoskeletons of arthropods by electrofiltration, an alternative method, thus reducing the downstream processing steps and costs. Experiments with different voltages and pressures were conducted in order to demonstrate the effect of both parameters on filtration kinetics. The concentration of the biopolymer was obtained by the average factor of 40 by applying an electric field of 4 V/mm and pressure of 4 bars. A series of analytical experiments demonstrated the lack of structural and functional changes in chitosan molecules after electrofiltration. These results, combined with the reduction of energy and processing time, define the investigated method as a promising downstream step in the chitosan production technology.
In der Natur konnte Chitosan bisher nur in den Zellwänden weniger Pilze (z. B. Mucorales) gefunden werden. Auf Grund struktureller Eigenschaften, der geringen Toxizität, Biokompatibilität und biologischen Abbaubarkeit besitzt Chitosan ein breites Anwendungspotenzial. Wegen entfallender Entsorgungs‐ und Umweltprobleme ist es zusätzlich als Rohstoff attraktiv. Chitosan wird bisher aus Chitin gewonnen, das überwiegend aus Schalenresten der Krabben‐ und Shrimpsherstellung stammt. Eigene Untersuchungen zeigen, dass sich Chitosan aus verschiedenen Pilzmycelien, die als Restprodukte bei biotechnologischen Prozessen (Enzymproduktion, Bierbrauerei) anfallen, besonders günstig gewinnen und aufarbeiten ließe. Unter Umständen ist sogar ein Fermentationsprozess mit Zygomycota rentabel, weil in der Zellwand dieser filamentösen Pilze bereits Chitosan in vivo enthalten ist und so auf den Aufarbeitungsschritt der Deacetylierung verzichtet werden kann. Analysen ergaben, dass Chitosan aus Pilzmycelien gleiche Eigenschaften wie Krabbenchitosan hinsichtlich des Deacetylierungsgrades und der Wasserlöslichkeit bei weniger Aufarbeitungsschritten zeigt. Außerdem lässt es sich protein‐ und schwermetallfrei herstellen, was für Anwendungen im medizinischen Bereich besonders wichtig ist.
Modelling of activated sludge processes is a commonly used technique to design and optimize wastewater treatment processes. Since wastewater and activated sludge is characterized by chemical oxygen demand (COD) measurements, units of state variables describing organic matter are expressed as equivalent amounts of COD. However, current procedures for measuring it have several drawbacks, including the production of hazardous wastes, so the utility of other variables for characterizing the organic load in modelling, such as total organic carbon (TOC), warrant re-evaluation. Other advantages of TOC over COD are that it provides matrix-independent analytical results and it can be readily measured online. Proposals for TOC-based models were made in the 1990s, but they seem to have sunk into obscurity. To re-assess the value of TOC for this purpose, we have recalculated the EAWAG module for Bio-P removal coupled to the Activated Sludge Model No. 3 on a TOC basis, and tested it against data acquired in batch experiments with four single carbon sources (acetate, glucose, citrate and casein). The batch test-based calibrations showed a good match with experimental data, following modifications of the model to account for the anaerobic volumes and retention times applied in the tests.
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