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Drivers Of On-Farm Performance Of Irrigation Water Management Practices: Empirical Evidence From Eastern Arkansas, Jacob Askey Dec 2019

Drivers Of On-Farm Performance Of Irrigation Water Management Practices: Empirical Evidence From Eastern Arkansas, Jacob Askey

Biological and Agricultural Engineering Undergraduate Honors Theses

Unsustainable agricultural practices are putting a strain on freshwater supplies in many parts of the world. More efficient irrigation techniques are going to be critical to feed a growing population. Data from the 2016 Arkansas Irrigation Survey was used to assess the adoption of three Irrigation Water Management (IWM) practices: multiple inlet rice irrigation, tailwater recovery, and storage reservoirs. Results indicate that these practices do not always lead to reported pumping time reductions, a proxy for water use savings. Large variations in pumping time reduction are observed across producers for all three practices. A Tobit model was used to examine ...


Functional Importance Of Lipin Phosphorylation, Stephanie Elizabeth Hood Dec 2019

Functional Importance Of Lipin Phosphorylation, Stephanie Elizabeth Hood

Theses and Dissertations

Highly conserved throughout evolution, lipins are dual functioning proteins found from yeast to humans. Functioning in the cytoplasm as phosphatidate phosphatase enzymes (PAP), lipins produce diacylglycerol that serves as a precursor for neutral fats and membrane phospholipids. Alternatively, nuclear lipins are responsible for the regulation of metabolic genes. Interestingly, both the mammalian lipin 1 paralog and the single Drosophila Lipin ortholog are highly phosphorylated proteins. Target of rapamycin (TOR) has previously been identified as one of the kinases that controls the subcellular localization of both lipin 1 and Drosophila Lipin. However, other serine and threonine kinases are predicted to be ...


Cytotoxicity And Mutagenicity Of Disinfection Byproduct Mixtures Formulated Using Free Chlorine And Monochloramine, Yonas Ghebremedhin Dec 2019

Cytotoxicity And Mutagenicity Of Disinfection Byproduct Mixtures Formulated Using Free Chlorine And Monochloramine, Yonas Ghebremedhin

Theses and Dissertations

Bioassays have been used extensively to assess various toxicity endpoints of drinking water disinfection byproducts (DBPs), but an emphasis on single compounds prevails. In this research, DBP mixtures were assessed using a cytotoxicity test with Chinese Hamster Ovary (CHO) cells and an Ames fluctuation test with Salmonella typhimurium TA-98 and TA-100 with and without S9 rat liver homogenate. Seven whole mixture DBP concentrates were formulated using reconstituted natural organic matter (NOM) extracts from the Upper Mississippi River using scaled disinfectant dosing protocols with free chlorine and monochloramine in the presence and absence of added bromide. DBPs were identified by gas ...


Influence Of Single And Multiple Histidine Residues And Their Ionization Properties On Transmembrane Helix Dynamics, Orientations And Fraying, Fahmida Afrose Dec 2019

Influence Of Single And Multiple Histidine Residues And Their Ionization Properties On Transmembrane Helix Dynamics, Orientations And Fraying, Fahmida Afrose

Theses and Dissertations

Since aromatic and charged residues are often present in various locations of transmembrane helices of integral membrane proteins, their impacts on the molecular properties of transmembrane proteins and their interactions with lipids are of particular interest in many studies. In this work, I used solid-state deuterium NMR spectroscopy in designed model peptide GWALP23 [GGALW(LA)6LWLAGA] with selective deuterium labels to addresses the pH dependence and influence of single and multiple “guest” histidine residues in the orientation and dynamic behaviors of transmembrane proteins. The mutations include Gly to His (G2/22 to H2/22), Trp to His (W5/19 to ...


Minimalistic Peptide-Based Supramolecular Systems Relevant To The Chemical Origin Of Life, Daniela Kroiss Sep 2019

Minimalistic Peptide-Based Supramolecular Systems Relevant To The Chemical Origin Of Life, Daniela Kroiss

All Dissertations, Theses, and Capstone Projects

All forms of life are based on biopolymers, which are made up of a selection of simple building blocks, such as amino acids, nucleotides, fatty acids and sugars. Their individual properties govern their interactions, giving rise to complex supramolecular structures with highly specialized functionality, including ligand recognition, catalysis and compartmentalization. In this thesis, we aim to answer the question whether short peptides could have acted as precursors of modern proteins during prebiotic evolution. Using a combination of experimental and computational techniques, we screened a large molecular search space for peptide sequences that are capable of forming supramolecular complexes with adenosine ...


Renal Specific Rnai Delivery By Fibrillar Nanoparticle Excipients, Sam Wong Sep 2019

Renal Specific Rnai Delivery By Fibrillar Nanoparticle Excipients, Sam Wong

All Dissertations, Theses, and Capstone Projects

RNA interference (RNAi) is a powerful tool to manipulate the phenotype of an organism by silencing the expression of specific genes and is viewed as a highly promising platform for treating undruggable targets and disorders where small molecule drugs and antibodies would fail. However, development of RNAi based therapies has faced major barriers including cellular and tissue-specific uptake of the Small Interfering RNA (siRNA). Utilizing different nanoparticles as RNAi excipients, cellular uptake and gene silencing potency can be greatly improved. The research in Dr. McDevitt groups has fibrillar carbon nanotubes (CNT) as carriers for siRNA for gene silencing in vitro ...


Developmental And Sex Modulated Neurological Alterations In Autism Spectrum Disorder, Azeezat Azeez Aug 2019

Developmental And Sex Modulated Neurological Alterations In Autism Spectrum Disorder, Azeezat Azeez

Dissertations

Autism Spectrum Disorder (ASD) was first described in 1943 by Dr. Leo Kranner in a case study published in The Nervous Child. It is a neurodevelopment disorder, with a range of clinical symptoms. According to the Diagnostic and Statistical Manual of Mental Disorders (DSM-5), used by clinicians to diagnose mental disorders, a child needs to have persistent social deficits, language impairments, and repetitive behaviors, that cannot be explained by neurological damage or intellectual disability. It is known that children diagnosed with ASD are often are developmentally delayed therefore alterations in the typical developmental trajectory should be a major factor in ...


Understanding Excitation Energy Quenching In Isia, Hui-Yuan Steven Chen Aug 2019

Understanding Excitation Energy Quenching In Isia, Hui-Yuan Steven Chen

Engineering and Applied Science Theses & Dissertations

Cyanobacteria are photoautotrophic organisms that contribute a significant amount of global primary productivity. They are found in freshwater, marine and even some extremely severe environments. Among those environments, iron deficiency is one of the most common stress conditions in cyanobacterial habitats. To survive, cyanobacteria have evolved and developed several strategies to alleviate the damage caused by iron deficiency.

Iron stress-inducible protein (IsiA) is a chlorophyll-binding membrane protein found in cyanobacteria grown in iron-deficient conditions. During the past decades, considerable effort has been put on understanding how IsiA functions to help cyanobacteria survive iron deficiency. It has been reported that IsiA ...


Metabolic Engineering Of Cyanobacteria For Production Of Chemicals, Po-Cheng Lin Aug 2019

Metabolic Engineering Of Cyanobacteria For Production Of Chemicals, Po-Cheng Lin

Engineering and Applied Science Theses & Dissertations

Concerns over the impact of climate change caused by CO2 emission have driven the research and development of renewable energies. Microbial production of chemicals is being viewed as a feasible approach to reduce the use of fossil fuels and minimize the impact of climate change. With recent advances in synthetic biology, microorganisms can be engineered to synthesize petroleum-based chemicals and plant-derived compounds. Cyanobacteria are photosynthetic prokaryotes that use only sunlight, CO2, and trace minerals for growth. Compared to other microbial hosts, cyanobacteria are attractive platforms for sustainable bioproduction, because they can directly convert CO2 into products. However, the major challenge ...


Understanding Excitation Energy Quenching In Isia, Hui-Yuan Steven Chen Aug 2019

Understanding Excitation Energy Quenching In Isia, Hui-Yuan Steven Chen

Engineering and Applied Science Theses & Dissertations

Cyanobacteria are photoautotrophic organisms that contribute a significant amount of global primary productivity. They are found in freshwater, marine and even some extremely severe environments. Among those environments, iron deficiency is one of the most common stress conditions in cyanobacterial habitats. To survive, cyanobacteria have evolved and developed several strategies to alleviate the damage caused by iron deficiency. Iron stress-inducible protein (IsiA) is a chlorophyll-binding membrane protein found in cyanobacteria grown in iron-deficient conditions. During the past decades, considerable effort has been put on understanding how IsiA functions to help cyanobacteria survive iron deficiency. It has been reported that IsiA ...


Metabolic Engineering Of Cyanobacteria For Production Of Chemicals, Po-Cheng Lin Aug 2019

Metabolic Engineering Of Cyanobacteria For Production Of Chemicals, Po-Cheng Lin

Engineering and Applied Science Theses & Dissertations

Concerns over the impact of climate change caused by CO2 emission have driven the research and development of renewable energies. Microbial production of chemicals is being viewed as a feasible approach to reduce the use of fossil fuels and minimize the impact of climate change. With recent advances in synthetic biology, microorganisms can be engineered to synthesize petroleum-based chemicals and plant-derived compounds. Cyanobacteria are photosynthetic prokaryotes that use only sunlight, CO2, and trace minerals for growth. Compared to other microbial hosts, cyanobacteria are attractive platforms for sustainable bioproduction, because they can directly convert CO2 into products. However, the major challenge ...


Investigation Of The Electrode Polarization Effect For Biosensor Applications, Anil Koklu Aug 2019

Investigation Of The Electrode Polarization Effect For Biosensor Applications, Anil Koklu

Mechanical Engineering Research Theses and Dissertations

My research focuses on electrokinetic transport. Particularly, in this dissertation, we focus on fabrication and testing of micro electrodes with nanostructured surfaces to minimize the electrode polarization (EP) effects for biosensor applications. In the first study, electrochemical deposition of gold nanoparticles on to planar gold electrodes was used to generate rough surfaces. Dendritic nanostructures that reduced EP up to two orders of magnitude was obtained by optimizing the deposition conditions. These structures also enhanced dielectrophoresis (DEP) response of our bio-chips, making them usable in physiological buffers. In further studies we discovered a universal scaling of EP in the frequency domain ...


Multi-Tap Extended Kalman Filter For A Periodic Waveform With Uncertain Frequency And Waveform Shape, And Data Dropouts, Justin Saboury Aug 2019

Multi-Tap Extended Kalman Filter For A Periodic Waveform With Uncertain Frequency And Waveform Shape, And Data Dropouts, Justin Saboury

Theses and Dissertations

Gait analysis presents the challenge of detecting a periodic waveform in the presence of time varying frequency, amplitude, DC offset, and waveform shape, with acquisition gaps from partial occlusions. The combination of all of these components presents a formidable challenge. The Extended Kalman Filter for this system model has six states, which makes it weakly identifiable within the standard Extended Kalman Filter network. In this work, a novel robust Extended Kalman Filter-based approach is presented and evaluated for clinical use in gait analysis. The novel aspect of the proposed method is that at each sample, the present and several past ...


Bioengineering Extracellular Matrix Scaffolds For Volumetric Muscle Loss, Kevin Roberts Aug 2019

Bioengineering Extracellular Matrix Scaffolds For Volumetric Muscle Loss, Kevin Roberts

Theses and Dissertations

Volumetric muscle loss overwhelms skeletal muscle’s ordinarily capable regenerative machinery, resulting in fibrosis and severe functional deficits which have defied clinical repair strategies. My work spans the design and preclinical evaluation of implants intended to drive the cell community of injured muscle toward a regenerative state, as well as the development of an understanding of the molecular responses of this cell community to biomaterial interventions. I demonstrate a new class of biomaterial by leveraging the productive capacity of sacrificial hollow fiber membrane cell culture; I show specifically that unique threads of whole extracellular matrix can be isolated by solvent ...


A Simulation Tool For Evaluating The Environmental Impacts Of Management Scenarios For Modern Broiler Production Systems, Martin Andrew Christie Aug 2019

A Simulation Tool For Evaluating The Environmental Impacts Of Management Scenarios For Modern Broiler Production Systems, Martin Andrew Christie

Theses and Dissertations

The purpose of this work is to provide a simulation tool that allows broiler production practitioners and researchers to simulate the effects of farm design and management practices on resource consumption and environmental impacts. This tool allows the user to design unique farms and simulates on farm processes required to raise broiler chicks to a marketable age. The use can input data such as farm location, broiler breed, flock size, ration type, barn dimensions, and climate control equipment specifications. The algorithms used to simulate broiler breed specific feed intake, broiler weight gain, and other on farm processes such as heating ...


Hybrid Fusion Protein For Inhibition Of Multiple Proteases For Chronic Wound Healing, Graham L. Strauss Jul 2019

Hybrid Fusion Protein For Inhibition Of Multiple Proteases For Chronic Wound Healing, Graham L. Strauss

Graduate Theses and Dissertations

Many diseases display a multitude of relevant factors that contribute to the persistence of the disease and difficulty treating it. The multifactorial characteristics of some diseases lead to the requirement of combination of treatments in order to restore health. The latter may necessitate the mixing of treatments, medications, and therapeutics to first halt the disease, then assist the human body in returning itself to a state of normality. For example, chronic wounds exhibit this multifactor characteristic in which there exist many factors that lead to the body’s inability to properly heal in a timely manner. This presents a further ...


Endogenous Force Transmission Between Epithelial Cells And A Role For Α-Catenin, Sandeep Dumbali Jul 2019

Endogenous Force Transmission Between Epithelial Cells And A Role For Α-Catenin, Sandeep Dumbali

Mechanical & Aerospace Engineering Theses & Dissertations

In epithelial tissues, epithelial cells adhere to each other as well as to the underlying extra-cellular matrix (ECM). E-cadherin-based intercellular junctions play an important role in tissue integrity. These junctions experience cell-generated mechanical forces via apparent adaptor proteins such as beta (β) catenin, alpha (α) catenin and vinculin. Abnormalities in these junctions may result in skin related diseases and cancers. Here, I devised methods to determine the endogenous intercellular force within cell pairs as well as in large epithelial islands. I further ascertained the factors that affect the level of inter-cellular tension.

Experiments with pairs of epithelial cells exogenously expressing ...


Tidalsim Senior Project Report, Kent Zhang, Colin Vandervoort Jun 2019

Tidalsim Senior Project Report, Kent Zhang, Colin Vandervoort

Computer Engineering

Throughout the course of this project, our team helped the Cal Poly Biological Sciences department refine an intertidal zone simulator. The aim of this device is to allow any marine biologist to easily simulate a vast range of tidal zones in order to test animal behaviors within these zones. Another goal of this project is to make each simulation tank independent from the others by using a single microcontroller to handle all inputs and outputs of the system as well as logging all relevant data. The current system is set up so that a separate microcontroller handles dissolved oxygen and ...


Compositional Optimization Of Amyloid-Graphene Oxide Nanohybrids For Biomaterials, Claire L. Drewery Jun 2019

Compositional Optimization Of Amyloid-Graphene Oxide Nanohybrids For Biomaterials, Claire L. Drewery

Materials Engineering

Amyloid nanofibrils are natural materials capable of self-assembling into precise structures with tunable functionalities, while exhibiting excellent mechanical properties. In combination with highly conductive graphene oxide (GO), the 1-D amyloid nanofibrils and 2-D nanosheets of GO can produce a robust and bio-functional nanohybrid, hypothesized to exhibit multi-domain functional properties useful for enzyme sensing, water purification, drug delivery, and tissue scaffolding applications. Here, we examine the properties of an amyloid-graphene oxide nanohybrid film made with amyloids derived from hen egg white lysozymes in an attempt to explore the diverse toolbox of amyloid derivatives and establish ideal fabrication methods and formulations of ...


Study Of Paper Microbial Fuel Cells For Use In On-Site Wastewater Testing, William A. Tolmasoff Jun 2019

Study Of Paper Microbial Fuel Cells For Use In On-Site Wastewater Testing, William A. Tolmasoff

Master's Theses and Project Reports

This study demonstrated a technique for fabricating simple, low-cost Paper Microbial fuel cells (PMFC’s) in the model of a previous study to, for the first time, produce voltage from wastewater effluent. The PMFC’s were created by stacking and gluing the main components of an MFC together: reservoir layer; anode; cation exchange membrane (CEM); air cathode. A wax printer was used to create the hydrophobic borders of the PMFC’s on filter paper, and graphite paint was applied to the paper to create the anode. The CEM’s considered were filter paper, wax, and Nafion, with Nafion being the ...


Carbonate Chemistry Characterization In A Low-Inflow Estuary With Recent Seagrass Loss, Jolie Higgins Jun 2019

Carbonate Chemistry Characterization In A Low-Inflow Estuary With Recent Seagrass Loss, Jolie Higgins

Master's Theses and Project Reports

Estuaries are dynamic environments that are strongly affected by natural variability, as well as direct and indirect anthropogenic impacts. A better understanding of the drivers of carbon fluxes and biogeochemical variability in estuarine systems is needed, particularly with the increasing threat of ocean acidification. Morro Bay in Central California is a small nationally protected estuary, with seasonally low freshwater inputs. Since 2007, the bay has experienced a significant loss of native seagrass, Zostera marina, which is an important component of the marine ecosystem. Because seagrass photosynthesis decreases carbon dioxide and increases oxygen in the water column, the loss of seagrass ...


Adaptive Paddle Board, Alexander Holthaus, Alexander Holthaus, Garrett Holmes, Garett Jones Jun 2019

Adaptive Paddle Board, Alexander Holthaus, Alexander Holthaus, Garrett Holmes, Garett Jones

Mechanical Engineering

This Final Design Review (FDR) document outlines the Adaptive Paddle Board senior project, done by four Mechanical Engineering Students at California Polytechnic State University and provides detail on the project and what the team has accomplished. The goal was to create a universally adaptive paddle board that can be used by the Central California Adaptive Sports Center for a wide range of persons with disabilities. This document highlights current research from patents and existing products, details regarding customer specifications, results from concept generation, the manufacturing and testing that went into the final design, and the process taken to get there ...


Fouling And Aging In Membrane Filtration : Hybrid Afm-Based Characterization, Modelling And Reactive Membrane Design, Wanyi Fu May 2019

Fouling And Aging In Membrane Filtration : Hybrid Afm-Based Characterization, Modelling And Reactive Membrane Design, Wanyi Fu

Dissertations

Membrane filtration has been extensively used in water and wastewater treatment, desalination, dairy making, and biomass/water separation. However, membrane fouling, aging and insufficient removal efficiency for dissolved organic matters remain major challenges for wider industrial applications. In order to tackle these challenges, this doctoral dissertation investigates mechanisms of membrane fouling and development of antifouling membrane filtration technologies. Specifically, four major research areas are explored: (i) nanoscale physicochemical characterization of the chemically modified polymeric membranes; (ii) quantitative modelling between membrane properties and membrane fouling and defouling kinetics; (iii) development of quantitative structure-activity relationships for membranes that undergo thermal and chemical ...


Electrochemically Reactive Membranes For Efficient Biomass Recovery, Pollutant Degradation And Commercialization, Likun Hua May 2019

Electrochemically Reactive Membranes For Efficient Biomass Recovery, Pollutant Degradation And Commercialization, Likun Hua

Dissertations

Micropollution in natural waters such as rivers and groundwater aquifers is a widespread problem that prevents these potentially potable sources from being used as drinking water. In the United States, approximately two-thirds of the over 1,200 most serious hazardous waste sites in the nation are contaminated with trichloroethylene (TCE), a potentially carcinogenic compound. Other emerging and environmentally persistent organic micropollutants include polyromantic hydrocarbons (PAHs), organophosphate flame retardants, endocrine disrupting compounds (EDCs), pesticides, herbicides, pharmaceuticals and personal care products (PPCPs). Membrane filtration is one of the most efficient separation processes widely used for water treatment and pollutant removal. However, traditional ...


Cloning A Fluorescent Fusion Protein For Detection And Localization Of The Gastrin Receptor, Vinh Le May 2019

Cloning A Fluorescent Fusion Protein For Detection And Localization Of The Gastrin Receptor, Vinh Le

Honors Projects and Presentations: Undergraduate

Pancreatic ductal adenocarcinoma (PDAC) is the fourth leading cause of cancer-related death worldwide. It is promoted by multiple cellular signaling events. Gastrin, a hormone that regulates several aspects of gastrointestinal physiology and maintenance of the gastrointestinal mucosa, is abnormally expressed in pancreatic cancer cells and promotes PDAC growth. Its effects are chiefly mediated through the seven membrane-spanning G protein-coupled receptor CCK2R. Upregulation of CCK2R and the cancer-associated splice variant, CCK2i4svR, have been shown to contribute to tumorigenicity, with the latter promoting increased tumor aggressiveness. Our lab has previously reported the development of monoclonal antibodies specific to CCK2R and CCK2i4svR. However ...


Coupled Correlates Of Attention And Consciousness, Ravi Varkki Chacko May 2019

Coupled Correlates Of Attention And Consciousness, Ravi Varkki Chacko

Engineering and Applied Science Theses & Dissertations

Introduction: Brain Computer Interfaces (BCIs) have been shown to restore lost motor function that occurs in stroke using electrophysiological signals. However, little evidence exists for the use of BCIs to restore non-motor stroke deficits, such as the attention deficits seen in hemineglect. Attention is a cognitive function that selects objects or ideas for further neural processing, presumably to facilitate optimal behavior. Developing BCIs for attention is different from developing motor BCIs because attention networks in the brain are more distributed and associative than motor networks. For example, hemineglect patients have reduced levels of arousal, which exacerbates their attentional deficits. More ...


Improved Orthopaedic Repairs Through Mechanically Optimized, Adhesive Biomaterials, Stephen Wheeler Linderman May 2019

Improved Orthopaedic Repairs Through Mechanically Optimized, Adhesive Biomaterials, Stephen Wheeler Linderman

Engineering and Applied Science Theses & Dissertations

Despite countless surgical advances over the last several decades refining surgical approaches, repair techniques, and tools to treat tendon and tendon-to-bone injuries, we are still left with repair solutions that rely on fairly crude underlying mechanical principles. Musculoskeletal soft tissues have evolved to transfer high loads by optimizing stress distribution profiles across the tissue at each length scale. However, instead of mimicking these natural load transfer mechanisms, conventional suture approaches are limited by high load transfer across only a small number of anchor points within tissue. This leads to stress concentrations at anchor points that often cause repair failure as ...


Modeling Mechanisms Behind Force Generation By Actin Polymerization, Seyed Fowad Motahari May 2019

Modeling Mechanisms Behind Force Generation By Actin Polymerization, Seyed Fowad Motahari

Arts & Sciences Electronic Theses and Dissertations

Actin polymerization is the primary mechanism for overcoming the large turgor pressure that opposes endocytosis in yeast. While generation of pushing forces by actin polymerization is fairly well understood, it is not clear how actin polymerization produces pulling forces. In order to understand this process, it is necessary to simulate polymerization of filaments having various types of interactions with the membrane. Since existing methodologies in the literature do not treat such problems correctly, we develop a thermodynamically consistent methodology for treating polymerization of filaments having arbitrary interaction potentials with the membrane. Then I perform stochastic simulations for a system of ...


A Ph-Sensitive Delivery System For The Prevention Of Dental Caries Using Salivary Proteins, Yi Zhu May 2019

A Ph-Sensitive Delivery System For The Prevention Of Dental Caries Using Salivary Proteins, Yi Zhu

Electronic Thesis and Dissertation Repository

Dental caries remains one of the most common chronic diseases worldwide. Salivary proteins such as histatins have demonstrated biological functions directly related to tooth homeostasis and prevention of dental caries. However, histatins are susceptible to the high proteolytic activities in the oral environment. Therefore, pH-sensitive chitosan nanoparticles (CNs) have been proposed as potential carriers to target major oral diseases that occur under acidic conditions (e.g. dental caries and dental erosion). Four different types of chitosan polymers were investigated and the optimized CNs successfully loaded histatin 3 and released it selectively under acidic conditions. Through loading the survival time of ...


A Feasibility Study Of Industrial Hemp On Virginia Commercial Poultry Production, Aaron Sloss May 2019

A Feasibility Study Of Industrial Hemp On Virginia Commercial Poultry Production, Aaron Sloss

Senior Honors Projects, 2010-current

This project sought to characterize industrial hemp as a potential competitive agricultural commodity for the state of Virginia. Specifically, industrial hempseed was evaluated as a feedstock for the commercial broiler (meat-chickens) industry in Virginia.

Data were gathered to identify both the advantages and disadvantages of industrial hempseed as a nutritional resource for broilers. Key nutritional compositions were identified, quantified, and compared to several common feed ingredients, such as soybean, wheat, corn, and sorghum. Hempseed (whole seed) was found to have a considerably high amino acid content, protein, fiber, and metabolizable energy – making it an attractive alternative to conventional feedstuffs.

However ...