Scaling biological pretreatment from the bench scale to the production scale may be more economical if unsterilized feedstock are used, however these allow for microbial competition from contaminates. An accurate and rapid method for identifying the desired biological pretreatment organism is necessary to confirm the presence of the desired organism when contaminates are morphologically similar to the target organism. Traditional methods, such as visual identification, sequencing, and selective plating can be time consuming and are sometimes still inconclusive. Based on methods described in the literature, plasmid DNA containing the marker genes gus (�-glucuronidase), LacZ, and gfp (green fluorescence protein) incorporated into the lignin-degrading basidiomycete Phanerochaete chrysosporium would result in a rapid genetic test for the desired organism. The presence of these genes can be confirmed either through an X-Gluc (cyclohexylammonia salt), X-Gal histochemical assay or observing the gfp’s fluorescence by a specially equipped confocal microscope. Each reporter systems will allow for rapid, reliable identification of the target species. This study will report on the success of the transformation methods in creating a transformed fungus to be used in the context of a large-scale fermentation operation. Additionally, a novel in-harvest lignocellulose feedstock biological pretreatment inoculation trial was performed comparing lignolytic performance between fungal inoculum application techniques. Optimization of carbohydrate availability for enhanced saccharification was determined by analyzing glucose release by treated and non-treated unsterilized switchgrass. This study also focused on identifying parameters to enhance saccharification efficacy at the farm-scale.

The following full-factorial study compared fungal activity on lignocellulosic biomass that was inoculated with three different amounts of fungus, and grown using three different airflow rates. These treatments were compared to a control which consisted of biomass that was not inoculated but was exposed to the same growth conditions in the environmental chamber. The objectives of the following experiment were to determine the inoculum density and airflow rate required to optimize Phanerochaete chrysosporium lignin degradation. Additionally, this study quantifies the saccharification yield from the pretreated switchgrass. The impact of substrate bulk density and substrate particle size on fungal growth were compared to determine if the particle size or the substrate bulk density has the predominant influence on the growth of the fungus, and subsequent pretreatment effectiveness quantified as an increase in glucose yields and lignin degradation. The particle size tests were controlled for bulk density; all three particle sizes were tested at a bulk density of 80 kg/m3. To test the density, three different bale densities were prepared controlling for particle size. The density tests were performed on small-scale bales made of 4 inch cut pieces of switchgrass compressed to the correct density. Therefore; density tests had the same particle size throughout all treatments, and particle size tests had the same density through all treatments. Carbohydrate accessibility post-pretreatment was examined through enzymatic saccharification and determination of glucose yields in the treatments and controls.

Compost bedded pack barns are a relatively new type of dairy housing system that is being implemented in Kentucky. Extensive research has been done on the composting of animal manure, however, little has been done on composting animal manure in place. One of the most concerning challenges is aeration. Improper aeration can cause system failure. The ability to quickly and accurately measure the oxygen concentration would allow researchers the ability to determine which methods of tillage/aeration are most effective in compost bedded pack barns. The research in this thesis focused on the development of a method for simultaneously testing oxygen concentrations at different locations and depth in compost in-situ. A probe was developed that vertically aligned Apogee Instruments oxygen sensors (SO-120) in order to generate an oxygen profile of the compost. The probe was used to test the effect of different tillage/aeration strategies in a composted bedded pack barn. The results indicated the probe was effective at measuring the oxygen concentrations in active compost tested in laboratory conditions and it was determined that there was a significant difference in oxygen concentration with respect to depth. However, when applied in the compost bedded pack barn, large amounts of variation occurred randomly in the data, causing no difference to be detected as a result of varying tillage aeration treatments.

Weathered sandstone materials have seen increased use in reclamation due to the wide-spread adoption of the Forest Reclamation Approach (FRA) in Appalachia. Runoff from these newly FRA reclaimed sites has the potential to adversely impact aquatic environments without fine sediment retention. To reduce the size and capital investment of settling ponds, flocculant utilization was investigated. Preliminary jar tests were conducted using composite weathered mine spoil samples acquired from a surface coal mine in eastern Kentucky. Four flocculants from the Magnafloc family of products were investigated during the initial screening-level testing. Experiments were conducted at three initial sediment concentrations (500 mg/L, 2,500 mg/L and 5,000 mg/L). A nonionic flocculant, Magnafloc 351, performed best, reducing total suspended sediment to below 50 mg/L. Large scale experiments confirmed that Magnafloc 351 was effective in reducing sediment concentrations. Jar tests were expanded to determine age and environmental effects on a Magnafloc 351 solution. Magnafloc 351 performance was slightly reduced after storage in a controlled building environment for 30 days and significantly decreased after 120 days. Magnafloc 351 solution exposed to UV and high heat (111°F) was ineffective after 30 days, while storage at 4°F and 36°F for 30 days did not adversely influence performance.

Because of high fuel costs and rising concern over controlling motor vehicle emissions, there has been a surge in the number of hybrid passenger vehicles on roads in recent years. This transition has not yet been seen with agricultural vehicles. With this in mind, this study created a test scheme to characterize and replicate agricultural loads, and design of a hybrid drivetrain that is suitable for agricultural purposes. Torque and power data were recorded from the controller area network of a tractor performing a baling operation. The recorded data was characterized using statistical and time series analyses, and converted into a simplified torque profile that could be run on a common type of dynamometer. The prototype series hybrid drivetrain was subjected to the simplified profile developed, and drivetrain efficiency was compared to the efficiency under constant load. The effect of battery pack, and engine size was also tested. On average, the prototype developed was not more efficient than a similarly sized standard geared vehicle, but there is significant room for further optimization.

Using the Cal Poly Irrigation Training and Research Centers (ITRC) drip/micro evaluation program, global, or system, DUlq is computed by combining the component DUlq values of: pressure variation, uneven spacing between emitters, unequal drainage and “other” causes. “Other” causes include plugging, wear and manufacturing coefficient. The program also computes what percentage of the non-uniformity is due to each component. Burt (2004) showed that over 95% of the non-uniformity is due to “Other” causes and pressure differences. This thesis looks at what specifically in those components is driving the non-uniformity by analyzing various equipment and field practices and their impact on the distribution uniformity. A regression analysis is used to analyze trends in distribution uniformity in an open environment. The results indicate that more information, specifically water quality, is needed to better analyze which components influence the distribution uniformity of a system.

The presence of moisture in the soil reduces its shear strength. After the rain or snowmelt, the high percentage of moisture in the subgrade of highway, causes the instability resulting into failure of the highway due to high water table. Therefore, it is essential to install a drainage system to remove the excess moisture from the subgrade of highways to avoid its failure. In the present study, six different soil samples have been studied to observe its failure by triaxial shear strength and corresponding moisture content was noted. The tension applied to remove moisture was converted to equivalent drain depth. The relationships were studied between shear strength versus drain depth and moisture content versus drain depth. From these relationships the design drain depth for different types of soils studied was recommended. The present study also suggest that the design drain depth for the highways can be approximated directly using water retention curve.

Microalgae have many desirable attributes as a renewable energy recourse. These include use of poor quality land, high yields, and it is not a food recourse. This research focusses on the energetic and environmental impact of processing microalgae into a renewable diesel. Two thermochemical bio-oil recovery processes are analyzed, pyrolysis and hydrothermal liquefaction (HTL). System boundaries include microalgae growth, dewatering, thermochemical bio-oil recovery, bio-oil stabilization, conversion to renewable diesel, and transportation to the pump. Two system models were developed, a small-scale experimental and an industrial-scale. The small-scale system model is based on experimental data and literature. The industrial-scale system model leverages the small scale system model with scaling and optimization to represent an industrial-scaled process. The HTL and pyrolysis pathways were evaluated based on net energy ratio (NER), defined here as energy consumed over energy produced, and global warming potential (GWP). NER results for biofuel production through the industrial-scaled HTL pathway were determined to be 1.23 with corresponding greenhouse gas (GHG) emissions of -11.4 g CO2 eq (MJ renewable diesel)-1. Biofuel production through the industrial-scaled pyrolysis pathway gives a NER of 2.27 and GHG emissions of 210 g CO2 eq (MJ renewable diesel)-1. For reference, conventional diesel has an NER of 0.2 and GHG emissions of 18.9 g CO2 eq MJ-1 with a similar system boundary. The large NER and GHG emissions associated with the pyrolysis pathway are attributed to feedstock drying requirements and combustion of co-products to improve system energetics. Process energetics with HTL and pyrolysis are not currently favorable for an industrial scaled system. However, processing of microalgae to biofuel with bio-oil recovery through HTL does produce a favorable environmental impact and a NER which is close to the breakeven point of one.

Algae can be used as a feedstock for agricultural fertilizers, livestock/poultry feeds, anaerobic digestion, and biofuel production. Regardless of the end product, water removal is necessary and difficult to do cost effectively. For each product the requirements for moisture content (or solids content) vary, such that a desirable water removal strategy would need to be adaptable to varying levels of water removal. Flocculation, with sedimentation and drying was evaluated as a possible strategy for algae dewatering. Anionic and nonionic flocculants are known to be ineffective at flocculating algal culture, which was confirmed for this case by electro-osmotic flow testing of the algae and jar tests with three flocculant charge types. Electrophoretic mobility of the algae indicated that it has a negative charge and no flocs were present in the jars. The effectiveness of the cationic flocculant was determined by measuring settling rates, supernatant turbidity, and filtration rates. Sedimentation and filtration rates of Scenedesmus acutus were measured with varying dosages (0-25 ppm) of a synthetic cationic polymeric flocculant. The results of this study should assist in predicting the time it takes to thicken algae at a concentration range of 0.4-1.0 g/L to a product at a concentration range of 15-250 g/L.

The temperature and moisture content of biomass feedstocks both play a critical role in minimizing storage and transportation costs, achieving effective bioconversion, and developing relevant postharvest quality models. Hence, this study characterizes the heat and mass transfer occurring within baled switchgrass through the development of a mathematical model describing the relevant thermal and physical properties of this specific substrate. This mathematical model accounts for the effect of internal heat generation and temperature-induced free convection within the material in order to improve prediction accuracy. Inclusion of these terms is considered novel in terms of similar biomass models. Two disparate length scales, characterizing both the overall bale structure (global domain) and the individual stems (local domain), are considered with different physical processes occurring on each scale. Material and fluid properties were based on the results of hydraulic conductivity experiments, moisture measurements and thermal analyses that were performed using the constant head method, TDR-based sensors and dual thermal probes, respectively. The unique contributions made by each of these components are also discussed in terms of their particular application within various storage and bioconversion operations. Model validation was performed with rectangular bales of switchgrass (102 x 46 x 36 cm3) stored in an environmental chamber with and without partial insulation to control directional heat transfer. Bale temperatures generally exhibited the same trend as ambient air; although initial periods of microbial growth and heat generation were observed. Moisture content uniformly declined during storage, thereby contributing to minimal heat generation in the latter phases of storage. The mathematical model agreed closely with experimental data for low moisture content levels in terms of describing the temperature and moisture distribution within the material. The inclusion of internal heat generation was found to be necessary for improving the prediction accuracy of the model; particularly in the initial stage of storage. However, the effects of natural convection exhibited minimal contribution to the heat transfer as conduction was observed as the predominate mechanism occurring throughout storage. The results of this study and the newly developed model are expected to enable the maintenance of baled biomass quality during storage and/or high-solids bioconversion.