University of South Florida

Western Autotroph Company LLC

Peter George Stroot

Raymond Anthony Morris

Matthew Raymond Cutter

Ryan Gates Toomey

Daniel Van Lim

Kelly M Leach

Joyce Marie Stroot

Samuel James DuPont, Jr

Lina Posso-Blandon

The present application describes methods and systems for measuring and controlling oxidative stress in animals and humans. The degree of oxidative stress can be measured directly by inducing all of the blood cells to produce excessive reactive oxygen species (ROS) by exposure to an elevated concentration sulfide or other ROS inducing chemical and measuring the fluorescence intensity of a fluorescent dye or color intensity of dye that reacts with ROS. Oxidative stress can be reduced by reducing dietary sulfur, consumption of a methanogenic probiotic, or apheresis methods to replace ROS-positive blood cells with normal blood cells. Plasma oxidative stress can be compared in venous and arterial blood samples to evaluate small vessel disease. Oxidative stress can be increased by increasing dietary sulfur or the use of an intravenous method that exposes blood cells to an elevated blood concentration of sulfide or other ROS inducing chemical.

Methods and systems for controlling oxidative stress

Methods and systems for controlling oxidative stress in humans and animals

The subject invention concerns materials and methods for detecting a target cell in a population. Methods of the invention comprise internally labeling cells via fluorescence in situ hybridization (FISH) using probes that target rRNA, followed by binding of capture antibodies targeted (CAT) for specific cell surface epitopes on the target cells. In one embodiment, the target cells are bacterial cells.

Methods and materials for capture antibody targeted fluorescent in-situ hybridization (CAT-FISH)

The present invention describes a method of optimizing COconcentration to increase the specific growth rate of Anammox bacteria and methanogens in wastewater and sludge treatment, as well as novel systems and methods of treating wastewater and sludge. The specific growth rate or doubling time of the Anammox bacteria and methanogens were determined to be sensitive to dissolved COconcentration. Optimizing dissolved COconcentration increases the specific growth rate of the Anammox bacteria, which may be used as an alternative biological nitrogen removal process for the treatment of domestic wastewater. In the method and system of treating sludge, the COstripper returns biogas with low COconcentration to the headspace of an anaerobic digester in order to lower the headspace COconcentration and therefore, the soluble COconcentration. The lower soluble COconcentration increases the specific growth rate of the methanogens for a more efficient anaerobic digestion process.

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The invention includes a method for the optimization of the soluble COconcentration in the aeration basin of an activated sludge system, which significantly improves the specific growth rate of the nitrifying bacteria. The result is a reduction in capital and energy costs for municipalities. The rate of nitrification is a product of the nitrifying bacteria biomass concentration and the specific growth rate of the bacteria. In the activated sludge system, the biomass concentration is maintained at high concentrations by reducing the wasting rate. The specific growth rate is a function of the ammonium concentration and the environmental conditions. Here, the inventors show that growth of nitrifying bacteria is inhibited when the soluble COconcentration is elevated beyond certain parameters. Elevated soluble COconcentration also reduces the pH, which also impacts the rate of growth of nitrifying bacteria.

The invention comprises two key components: dielectrophoresis (DEP) and reversible binding surfaces. DEP has become an important tool for trapping dielectric particles. Moreover, DEP can manipulate cell movement as dictated by the intrinsic dielectric constant of the cell without modification. DEP therefore provides a mechanism by which to force targets in a flow channel to a reversible binding surface. By building selectivity into the binding surface, the capacity to choose which targets can be held after the dielectric field is turned off, providing a separation strategy that does not suffer from fouling issues, as large foulants can freely pass over the surface through the flow channel.

Microbial cell and particle selection system and method of use

Treatment systems are described which uncouple the solids retention time (SRT) of the recycling bacteria from the wasting bacteria. Elevated recycling bacteria SRTs increase concentrations of nitrifying bacteria in the aeration basin and Anammox bacteria in the anoxic basin, improving nitrification and anaerobic ammonium oxidation. Fluorescence in situ hybridizations were used to identify and determine the abundance of nitrifying bacteria and Anammox bacteria (recycling bacteria) in freshly settled sludge of biological nutrient removal (BNR) secondary clarifiers. An uneven distribution was observed for recycling bacteria in two BNR systems. Settling patterns suggest microcolony formation processes are sensitive to the internal recycle rate, which allow for rapid recycling bacteria microcolony growth by increasing circulation of flocs through the aeration and anoxic basins. Uncoupling the SRT enhances aeration and/or anoxic basin utilization and reduces the overall hydraulic residence time.

System for solids retention time uncoupling by selective wasting of sludge

Fluorescence in situ hybridizations were used to identify and determine the abundance of nitrifying bacteria and Anammox bacteria (recycling bacteria) in freshly settled sludge of biological nutrient removal (BNR) secondary clarifiers. An uneven distribution was observed for recycling bacteria in two BNR systems. Settling patterns suggest microcolony formation processes are sensitive to the internal recycle rate, which allow for rapid recycling bacteria microcolony growth by increasing circulation of flocs through the aeration and anoxic basins. Sludge containing high levels of recycling bacteria are selectively collected and separated from the remaining sludge, uncoupling the solids retention time (SRT) of the recycling bacteria from the wasting bacteria. Elevated recycling bacteria SRTs increase concentrations of nitrifying bacteria in the aeration basin and Anammox bacteria in the anoxic basin, improving nitrification and anaerobic ammonium oxidation. Uncoupling the SRT enhances aeration and/or anoxic basin utilization and reduces the overall hydraulic residence time.

Solids retention time uncoupling by selective wasting of sludge

The present invention provides a method for measuring the specific rate of ribosome synthesis for a distinct cell population, such as a distinct microbial population. For an actively growing (or non-growing) culture, the specific rate of ribosome synthesis is identical to the specific growth rate of the culture. With the method of the invention, researchers will be able to measure the specific growth rate of distinct cell populations in mixed cultures, such as biological reactor systems or environmental samples. In addition, the method of the invention provides the ability to identify members of a distinct cell population that are rapidly growing.

Method for determining the specific growth rate of a distinct cell population within a non-homogeneous system

The present invention pertains to a molecular biology-based method and kit for measuring the specific growth rate (or cell doubling time) of distinct microbial populations. The method and kit can be used to analyze mixed culture samples that have been exposed to chloramphenicol or other protein synthesis inhibitors for defined times. In a preferred embodiment, the method of the invention (also referred to herein as FISH-RiboSyn) is an in situ method that utilizes fluorescence in situ hybridization (FISH) with probes that target: (1) the 5' or 3&#x2032; end of precursor 16S rRNA; or (2) the interior region of both precursor 16S rRNA and mature 16S rRNA. Images can be captured for a defined exposure time and the average fluorescent intensity for individual cells can be determined. The rate of increase of the whole cell fluorescent intensity is used to determine the specific growth rate. The method of the invention can be attractive for rapidly measuring the specific growth rate (or cell doubling time) of distinct microbial populations within a mixed culture in industries such as environmental systems (water and wastewater treatment systems), bioremediation (optimization of conditions for microbial growth), public health (identification of rapidly growing infectious microbes), and homeland security (identification of rapidly growing bioterrorism agents).

Method for determining the specific growth rate of distinct microbial populations in a non-homogeneous system

A method of stimulating nitrification at low SRT by elevating pCOduring aeration is disclosed. The improvement on solids settling performance when elevated pCOwas supplied after 2 hours within the React cycle is consistent with the previous results that identified inorganic carbon as a potential remedy to poor settling and bulking sludge problems in activated sludge systems. Elevated pCOincreases the concentration of carbon dioxide and lowers the pH, which improve nitrification. The specific growth rate of nitrifying bacteria is sensitive to pCO, pH, and dissolved oxygen (DO). The DO is a function of the aeration rate. Elevating the pCOand lowering the aeration rate provides conditions for nitrification rates that are comparable to conventional systems. However, the lower aeration rate yields significant energy cost savings.

Carbon dioxide stimulation of nitrification in activated sludge reactors

An anaerobic digestion process for the treatment of domestic wastewater sludge that requires less space and funding to construct. Sludge to be treated is combined with recycled anaerobic digester sludge to form a blended sludge. The recycled anaerobic digester sludge provides a source of microorganisms necessary to initiate the breakdown of organic matter in the sludge to be treated. The sludge is then concentrated to increase total solids content to about 10-20%. Excess liquid is removed from the concentrated sludge. The concentrated sludge is then digested in an anaerobic reactor system such as a plug-flow reactor. Some benefits of the system's reduced volume, as a result of concentration of the sludge, include elimination of the necessity of substantially continuous stirring and the new possibilities for the types of construction to be used for the reactor. In addition to the reduced cost of the process itself, the process creates biogas that can be used to offset energy requirements for the process.

Growing community of inventors

Lutz, FL, United States of America

Peter George Stroot