Microscope images of fluctuating biopolymers contain a wealth of information about their underlying mechanics and dynamics. However, successful extraction of this information requires precise localization of filament position and shape from thousands of noisy images. Here, we present careful measurements of the bending dynamics of filamentous (F-)actin and microtubules at thermal equilibrium with high spatial and temporal resolution using a new, simple but robust, automated image analysis algorithm with subpixel accuracy. We find that slender actin filaments have a persistence length of approximately 17 microm, and display a q(-4)-dependent relaxation spectrum, as expected from viscous drag. Microtubules have a persistence length of several millimeters; interestingly, there is a small correlation between total microtubule length and rigidity, with shorter filaments appearing softer. However, we show that this correlation can arise, in principle, from intrinsic measurement noise that must be carefully considered. The dynamic behavior of the bending of microtubules also appears more complex than that of F-actin, reflecting their higher-order structure. These results emphasize both the power and limitations of light microscopy techniques for studying the mechanics and dynamics of biopolymers.
A key future challenge of domestic wastewater treatment is nutrient recovery while still achieving acceptable discharge limits. Nutrient partitioning using purple phototrophic bacteria (PPB) has the potential to biologically concentrate nutrients through growth. This study evaluates the use of PPB in a continuous photo-anaerobic membrane bioreactor (PAnMBR) for simultaneous organics and nutrient removal from domestic wastewater. This process could continuously treat domestic wastewater to discharge limits (<50 mgCOD L(-1), 5 mgN L(-1), 1.0 mgP L(-1)). Approximately 6.4 ± 1.3 gNH4-N and 1.1 ± 0.2 gPO4-P for every 100 gSCOD were removed at a hydraulic retention time of 8-24 h and volumetric loading rates of 0.8-2.5 COD kg m(3) d(-1). Thus, a minimum of 200 mg L(-1) of ethanol (to provide soluble COD) was required to achieve these discharge limits. Microbial community through sequencing indicated dominance of >60% of PPB, though the PPB community was highly variable. The outcomes from the current work demonstrate the potential of PPB for continuous domestic (and possibly industrial) wastewater treatment and nutrient recovery. Technical challenges include the in situ COD supply in a continuous reactor system, as well as efficient light delivery. Addition of external (agricultural or fossil) derived organics is not financially nor environmentally justified, and carbon needs to be sourced internally from the biomass itself to enable this technology. Reduced energy consumption for lighting is technically feasible, and needs to be addressed as a key objective in scaleup.
Purple phototrophic bacteria (PPB) have been recently proposed as a key potential mechanism for accumulative biotechnologies for wastewater treatment with total nutrient recovery, low greenhouse gas emissions, and a neutral to positive energy balance. Purple phototrophic bacteria have a complex metabolism which can be regulated for process control and optimization. Since microbial processes governing PPB metabolism differ from traditional processes used for wastewater treatment (e.g., aerobic and anaerobic functional groups in ASM and ADM1), a model basis has to be developed to be used as a framework for further detailed modelling under specific situations. This work presents a mixed population phototrophic model for domestic wastewater treatment in anaerobic conditions. The model includes photoheterotrophy, which is divided into acetate consumption and other organics consumption, chemoheterotrophy (including simplified fermentation and anaerobic oxidation) and photoautotrophy (using hydrogen as an electron donor), as microbial processes, as well as hydrolysis and biomass decay as biochemical processes, and is single-biomass based. The main processes have been evaluated through targeted batch experiments, and the key kinetic and stoichiometric parameters have been determined. The process was assessed by analyzing a continuous reactor simulation scenario within a long-term wastewater treatment system in a photo-anaerobic membrane bioreactor.
Low wastewater temperatures affect microbial growth rates and microbial populations, as well as physical chemical characteristics of the wastewater. Wastewater treatment plant design needs to accommodate changing temperatures, and somewhat limited capacity is a key criticism of low strength anaerobic treatment such as Anaerobic Membrane Bioreactors (AnMBR). This study evaluates the applicability of an alternative platform utilizing purple phototrophic bacteria for low temperature domestic wastewater treatment. Two photo-anaerobic membrane bioreactors (PAnMBR) at ambient (22 °C) and low temperatures (10 °C) were compared to fully evaluate temperature response of critical processes. The results show good functionality at 10 °C in comparison with ambient operation. This enabled operation at 10 °C to discharge limits (TCOD < 100 mg L(-1); TN < 10 mg L(-1) and TP < 1 mg L(-1)) at a HRT < 1 d. While capacity of the system was not limited, microbial community showed a strong shift to a far narrower diversity, almost complete dominance by PPB, and of a single Rhodobacter spp. compared to a more diverse community in the ambient reactor. The outcomes of the current work enable applicability of PPB for domestic wastewater treatment to a broad range of regions.
Concentrated wastewaters from agricultural industries represent a key opportunity for the upcycling of organics, nitrogen and phosphorus to higher value products such as microbial protein. Phototrophic or photosynthetic microbes very effectively capture input organics and nutrients as microbial protein. This study compares purple phototrophic bacteria (PPB) and microalgae (photosynthesis) for this purpose, treating real, high strength poultry processing wastewater in continuous photo bioreactors utilising infrared (IR) and white light (WL) respectively. Both reactors could effectively treat the wastewaters, and at similar loading rates (4 kgCOD md). The infrared reactor (IRR) was irradiated at 18 W m and the white light reactor (WLR) reactor at 1.5-2 times this. The IRR could remove up to 90% total chemical oxygen demand (TCOD), 90% total nitrogen (TN) and 45% total phosphorus (TP) at 1.0 d hydraulic retention time (HRT) and recover around 190 kg of crude protein per tonne of influent COD at 7.0 kWh per dry tonne light input, with PPB dominating all samples. In comparison, the WLR removed up to 98% COD, 94% TN and 44% TP at 43-90% higher irradiance compared to the PPB reactor. Microalgae did not dominate the WLR and the community was instead a mix of microbes (algae, bacteria, zooplankton and detritus - ALBAZOD) with a production of approximately 140 kg crude protein per tonne influent COD.
Purple phototrophic bacteria (PPB) use infrared light to generate microbial energy, with electron, carbon and nitrogen supplied chemically. They have broad applicability for resource recovery from wastes and wastewater, as they have potential value as microbial product and very high growth yields on substrate. Although PPB processes are gaining relevance in the aforementioned industries, reactor design, particularly effective consideration of light in chemical kinetics and hydraulics is a critical issue. This thesis aims to assess key limitations and define their physical behaviour mathematically. The first limitation was that there was no mixed culture PPB process model describing the behaviour of PPB in the presence of ammonia, organic matter and other nutrients. The mixed population models of the International Water Association (IWA) provide a good framework for process modelling and control. As such, a process model for a mixed-culture PPB system was developed based on five key processes: photoheterotrophic growth, photoautotrophic growth, chemoheterotrophic growth, hydrolysis/fermentation, and decay. This model was developed as a set of ordinary differential equations varying in time and lumped in space (as for the IWA models). The second limitation is that the radiative field has not been considered. This can vary spatially and temporally. The radiative field in a PPB system attenuates sharply, which influences the growth domains. A CFD modelling framework was therefore developed in OpenFOAM to describe the spatial variations and interactions between biomass growth, the flow field and the radiative field. It was found that lumped modelling approaches and distributed parameter approaches differed in results based on different reactor domains and behaviours. The deviation from lumped parameter behaviour was greater for a cylindrical stirred reactor than for a flat plate reactor. Therefore, spatial considerations are necessary in the design phase of a photobioreactor. Finally, biofilms are a critical aspect in PPB growth, and a coupled biofilm radiative transfer model has not been previously considered. The model was thus extended to include biofilm formation of a mixed PPB system in the presence of a spatially varying radiative field. A volume-of-fluid approach was used as a basis for this model, considering three particulate species (phototrophic bacteria, biodegradable particulate matter, and inert particulates). The radiative-solid-liquid coupling overall was found to be critical in operation, with traditional lumped parameter approaches to design and analysis not well suited to the emerging challenges of mixed culture photo-bio systems. These aspects should be further considered through model based analysis and experimental validation for future system design.
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