Many studies have been conducted on tilapia throughout its commercial expansion; however potential antagonisms involving branched amino acids (BCAA), particularly their effects on growth and nutrient utilization, have not been fully assessed. Therefore, an 8-week feeding trial was conducted to access the effects of excess dietary BCAA [leucine (Leu) and isoleucine (Ile)] on growth, feed utilization, and blood parameters of juvenile Nile tilapia (Oreochromis niloticus). Three diets were formulated to be isonitrogenous (40% crude protein), isolipidic (10% lipid), and to contain different levels of the two essential amino acids. A basal diet (Basal) was designed to fulfil all dietary requirements of Nile tilapia. Two additional test diets (Leu+ and Ile+) were designed to contain Leu and Ile at an excess of 2.5% from Basal levels, respectively. Each diet was fed twice daily to apparent satiation to triplicate groups of 20 juvenile Nile tilapia (4.0 g initial mean weight) stocked into 110-L glass aquaria operating as a recirculating aquaculture system. After 8 weeks of feeding, we found no significant effects of excess Leu or Ile on the overall production performance and blood parameters of the experimental fish, indicating no antagonistic interactions. Therefore, since a 2.5% excess of Leu or Ile is very unlikely to occur in practical diet formulations for Nile tilapia, no upper limits of either amino acid in feeds for this species are necessary.

J. Clark, B. Webster, B. Chadwick, M. Mercer, I.P. Handayani, and B. Parr
Murray State University, Hutson School of Agriculture, Kentucky, USA

Abstract
Over time, managing different crops has caused soils to change their characteristics. The study includes a comparison between soils found in five different locations. Using the bulk density, water holding capacity, soil organic matter, soil pH, soil water content at field capacity, and soil porosity, this study aims to distinguish how continuously using the same cropping systems will affect soils over an extended period of time. Soil samples were collected from Christian County, Kentucky, Caldwell County, Kentucky, Calloway County, Kentucky, and Madison County, Illinois on August 29th 2017. The soil samples collected include silt loams and sandy silt loams that are in use for horse pasture, raising dark tobacco, conservation reserve program, gardening, and a park. From each location, three undisturbed soil samples and three disturbed samples were taken from the depth interval of 0 to 6.5 cm. The results show that the growth of different plants species can alter soil physical and chemical conditions under different cropping systems. The detail results and further discussion of this study will described in the poster. In general, the change of soil characteristics due to continuous cultivation was positive and negative depending on the type of land cover.

Keywords: Acidity, Bulk Density, Illinois, Kentucky, Organic Matter, Soil Water

Autumn olive is widely available in reclaimed coal mine fields in Eastern Kentucky. It produces small edible berries that can be made into jams. The objective of this project was to characterize the drying behavior of autumn berries. Juices extracted from the berries and measured for the sugar content with a Brix meter. Batches of berries were dried at 65 F in a dehydrator. The weight of the berries were measured at hour 0, 3, 6, 12, 15, 18, and 21 after the drying was started. The sugar content of the berries ranged from 10% to 14%. Weight loss of the berries was the largest during the initial 6 hours. The weight became constant after 18 hours, indicating the drying was complete. The dry matter content of the berries was 31%. Examination of the dried berries showed the seeds accounted for most of the dried weight.

Kentucky is a state which has a relatively small industry for the production of nursery, greenhouse, and floriculture crops. Ornamental horticulture is perhaps one of the most neglected areas in sustainable agriculture. Heavy use of chemicals, both pesticides and fungicides, is rampant, and production of ornamentals requires considerable energy and labor inputs. While the public may be aware of practices such as rain gardening and xeriscaping, some of these can be difficult to directly incorporate in existing gardens. The current project was undertaken to develop common garden plants that are cold-hardy and require lower energy input, thereby reducing overall carbon foot print. One group of horticulturally important ornamentals is the genus Begonia, which contains over 1,500 tropical or subtropical species along with a few cold-hardy species. Another popular genus, Zinnia, belongs to the subtribe Heliantheae, which also contains popular genera such as Echinacea and Helainthus. This genus itself contains about 20 species, distributed from the western U.S. to South America. Z. violacea, the common Zinnia, has been crossed with another species of Zinnia, Z. angustifolia to develop Z. maryalandica, a hybrid with a resistance to powdery mildew. The objectives of the current project were to impart cold hardiness into Begonia through interspecific hybridization with cold-hardy species (B. grandis), and to introduce perenniality into Zinnia violacea.

Distinguishing Differences of Soil Physical Properties
in Irrigated and Non- Irrigated Corn-soybean Rotation and Pasture Fields

D. Thomas, H. Franklin, C. Raymond, J. Fleming, I. P. Handayani and B. Parr
Murray State University, Hutson School of Agriculture, Murray, KY, USA.

Abstract
Land management practices such as irrigation, crop rotation and crop type affect soil properties. The objective of this study is to determine the effect of common land management practices in western Kentucky including non-irrigated, irrigated, and pasture ground on selected soil properties. Undisturbed and disturbed soil samples were collected from two fields in each of the three locations, from the depth of 1-3 inches and 3-6 inches. The fields in Princeton, KY were double crop soybeans in a no-till operation. In Simpson County, the fields were planted in corn, on a no-till operation and the field in Ohio County, KY was a non-irrigated pasture for grazing purposes. Soil samples in Princeton, KY were taken on September 8th and in Simpson County, KY, samples were collected on September 16th, 2017 due to flooding issues. In addition, soil samples in Ohio County, KY were taken on September 9th, 2017. All samples were analyzed for soil water holding capacity, soil water at field capacity, soil organic matter, bulk density, porosity and soil acidity level. Data from this study will be described in detail on the poster. The results will show if irrigation practices can improve soil conditions in a corn-soybean crop rotation.

Keywords: Crop rotation, Irrigated soil, Pasture, Soil physical properties, Western Kentucky

Lotus is grown for foods in China and other Asian countries because its seeds and roots are all edible and its flowers can be used for ornamental purposes. It is possible to grow lotus plants in containers. However, management of the plants such as fertilization is not well known. The objective of this study was to determine the effects of fertilization rate on the growth performance of lotus plants. Three varieties of lotus were tested with three levels of fertilization were tested in an experiment with 36 plastic containers (18 gallon volume) on a private farm in Nicholasville, KY. The containers were filled with a soil and compost mix (50:50) to 40 cm deep and rain water covering the soil and 10 cm below the rim of the containers. One root tube was planted into each container on May 1, 2017. Four levels of fertilizer (10-10-10), 10g, 20g, 40g or 80g was applied to each container according to a 3 x 4 factorial design one month after the tubes were planted into the containers. The number of floating leaves and standing leaves were counted each month. The number of standing leaves and total number of leaves increased from containers with 10g fertilizers to containers with 20g fertilizers. However, the number of leaves tended to become lower in containers with 40g or 80g fertilizers. In fact, lotus plants died in several containers with 80g fertilizers. The results indicate that over fertilization could lead to loss of lotus plants. fertilizing at 20g per container will be helpful.

Impact of Soil-Set®, Grain-Set®, and Liqui-Plex® Formulations on Hot Pepper
Yield, Plant Morphology and Fruit Quality

ABSTRACT
The consumption of hot pepper has increased during the last decade due to the increased consumption of ethnic populations and a greater interest in ethnic foods in the U.S. Environmentally and economically viable agriculture requires the use of cultivation practices and agrochemicals that maximize efficacy, crop nutritional composition and yield. The impact of Soil-Set® (a soil amendment that contains natural enzymatic compounds and balanced nutrients), Grain-Set® (a foliar fertilizer), and Liqui-Plex® Bonder WP (a foliar fertilizer that contains minerals complexed with amino acids) on pepper yield and fruit quality characteristics (fruit length, width, wall thickness, and early ripening) and plant morphological characteristics (shoot and root length) were investigated. A randomized complete block design (RCBD) experiment was conducted at Kentucky State University Research and Demonstration Farm. Four treatments (Soil-Set, Grain-Set, Liqui-Plex, and control) were replicated four times. Pepper (Capsicum annuum var. Georgia Flame) seedlings of 52 d old were planted and drip-irrigated as needed. At harvest, mature red fruits were collected, weighted, counted, and their quality characteristics (length, width, and wall-thickness) were measured. Results revealed that the plants treated with Soil-Set and Grain-Set formulations during the growing season produced the greatest fruit length (12.2 cm). Whereas, plants treated with Liqui-Plex formulation produced the greatest yield and greatest number of ripe fruits compared to the other treatments. These results could promote the use of these formulations in growing pepper and other vegetables in Kentucky and other states.

INTRODUCTION
Farmers' markets have been the most popular markets for local fresh produce. As more consumers increasingly shop online and fruits and vegetables being available in the online market, it is vital to investigate whether the online shoppers attend farmers' markets in their communities. In this study, we identify characteristics of whose who attends and who do not. We further present reasons for not attending.
SPECIFIC STUDY OBJECTIVES
1. Explain the frequency of attendance at farmers' markets among online shoppers,
2. Describe the overall experience at farmers' markets among online shopper attendees,
3. Describe reasons why some online shoppers do not attend farmers' markets.
METHODOLOGY
This study uses data collected from a stratified randomly selected sample of 1,205 online shoppers in 15 states of the South region of the U.S. Data were collected in 2016 using a survey emailed to the Qualtrics actively managed market research panels and those using social media such as Facebook and Twitter. Qualtrics is a professional survey software provider that offers sophisticated and advanced online data collection tools combined with respondent panels.
KEY FINDINGS
On average, the majority of online shoppers attend farmers' markets occasionally. Almost all online shoppers who attend farmers' markets indicated that they had at least a satisfactory overall experience. Major reasons for those who do not attend farmers' markets are (a) unawareness for existence of the markets in the area, (b) inconvenient hours of operations, and (c) inconvenient place (limited parking, long distance, do not like the location, and not a one-stop shopping destination).

Soil Distinguishing Factors in Pasture and Irrigated Soils
in Tennessee, Kentucky, and Illinois

S Fulkerson, L Williams, B Remmert, J Reddick, I. P. Handayani, and B. Parr
Murray State University, Hutson School of Agriculture, Murray, Kentucky USA

Abstract

Pasture and cropland soils have different properties which influence land productivity. The objectives of this study are to observe (1) The variations of soil properties in three states; (2) access the soil properties of cropland and pasture soil; (3) and compare the soils in terms of health and productivity. The undisturbed and disturbed soil samples from pasture and irrigated crop land were collected from Tennessee, Kentucky, and Illinois. These soils were sampled on September 2-3, 2017. Soil samples were analyzed for soil water holding capacity, soil water content at field capacity, bulk density, soil porosity, soil organic matter, and soil pH. In addition, soil map using web soil survey will be utilized to do a correlation between soil properties and soil type. The data will be presented on the poster. This research will help to better understand the properties of soils in agricultural production.


Keywords: Acidity, Bulk Density, Organic Matter, Water Content

Soil Properties in Varying Crop and Non-Crop Areas of Calloway County, Kentucky

Zack Eells, Clay Smotherman, Canaan Wring, Connor Moore, Iin Handayani, and Brian Parr
Murray State University, Hutson School of Agriculture, Kentucky, USA
Abstract
Cropping practices leading to loss of soil organic matter thus can alter other soil properties. The purpose of this study was to determine the impact of crop and non-crop areas on soils. Disturbed and undisturbed soil samples were collected from different fields of corn, soybeans, tobacco, pasture, and wooded areas in the Southwest portion of Calloway County, Kentucky on September 8, 2017. The properties observed include soil water holding capacity, soil water content at field capacity, bulk density, soil porosity, soil organic matter, and soil pH. All the data will be analyzed for means and standard error to observe the significant difference among the fields. The detail results will be explained on the poster. The data from this study can be used to quantify the physical and chemical changes of soils after cultivating the wooded areas for grain crops and tobacco.

Keywords: Acidity, Calloway County Kentucky, Soils, Corn-Soybeans-Tobacco, Wooded areas

Abstract

Cropping systems in Kentucky are strongly dominated by corn, tobacco, soybeans and wheat. Various crop management practices affect soil properties resulting in different functional quality of the soil to support crop growth. Crop rotation and tillage practices influence soil properties, and understanding the effect of these practices is essential to maintaining optimal soil environment. Therefore, the objective of this research is to determine the effects of various common cropping practices in western Kentucky on soil chemical and physical properties. In addition, soil samples from pasture, wood, and garden practices will be collected to use as a reference for comparison among crop fields. The study sites were selected from Caldwell County. There was six cropping systems identified, such as Monoculture systems (MS), Crop rotation Systems (CRS), Tobacco Cropping Systems (TCR), Corn/Soybean – cover crop (CC), Corn/Soybean-wheat (CSW), and three undisturbed fields (pasture, forest, and garden). Soils were sampled on October 1st, 2017. Undisturbed and disturbed samples were collected from the fields at depths of 0-7 cm and 7-15 cm. Undisturbed samples were collected using soil core (ring sample), while disturbed samples were taken using hand trawl. The samples were kept in the refrigerator until analysis. The undisturbed soil samples were used to analyze bulk density, porosity, macroporosity, and soil water holding capacity. The disturbed samples were used to analyze soil organic matter (SOM), soil pH and water stable aggregates (WAS). Soil compaction will also be directly measured in the field using penetrometer. Data from this study will be analyzed using the ANOVA. The least significant differences (LSD) values will be reported at a level (α)= 0.10. The correlation will be used to evaluate the relationship between SOM and other soil properties. The detail results will be described in the poster.

A field trial was established at the University of Kentucky South Research Farm. Eggplant, Solanum melongena var. Epic seedlings of 60 d old were planted in raised freshly tilled soil at eighteen inch in row spacing. Each plot was 4 × 10 feet2 and the entire study area contained 42 plots (3 replicates × 14 treatments). The experiment was replicated in a randomized complete block design (RCBD) with the following treatments: 1) control (untreated soil); 2) sewage sludge; 3) horse manure; 4) chicken manure; and 5) vermicompost (worm castings), 6) organic fertilizer (Nature Safe 20:2:8); and 7) inorganic fertilizer (Southern State 19:19:19). Each of the 7 treatments was also mixed with 10% (w/w) biochar obtained from Wakefield Agricultural Carbon (Columbia, MO) to make a total of 14 treatments. Fruits of eggplant were harvested during the growing season and graded according to the USDA guidelines into Fancy, US No.1, US No. 2, and Culls (unmarketable fruits). Results revealed that fruits of harvest 1 plants treated with inorganic fertilizer produced the greatest average weight of culls (978 g/plot) compared to HM and NM treatments (505 and 477 g/plot, respectively). In harvest 2, the number of fruits (29 and 22 fruits/plot) and weight of fruits (12 and 8 kg/plot) obtained from vermicompost treatments were significantly greater compared to other soil treatments. Overall five harvests, weight of fruits (13.7 kg/plot) obtained from inorganic treatments were significantly greater compared to vermicompost, SS, organic, and CM treatments.

Alfalfa is known as “queen of the forages,” due to high yield and high quality. Normally, forage quality decreases with maturity due to lignification of the plant cell walls, but a genetically enhanced alfalfa type, HarvXtra, has been developed which contains about 15% less lignin. This improved quality can translate into higher milk and meat production in cattle. The objective of this project was to determine forage quality, yield and botanical composition of low lignin and standard alfalfa in binary mixtures with cool season grasses. A randomized complete block design study was established at the University of Kentucky north farm with pure stands of low lignin and standard alfalfa in combination with the cool season grasses orchardgrass, meadow fescue and festulolium. The study was arranged in split blocks with half of the area harvested at bud stage and half at the flowering stage. The results showed that low lignin binary mixtures were lower yielding than standard binary mixtures, but pure stands of the two alfalfa types were not significantly different. Standard alfalfa showed higher alfalfa/grass composition in comparison to low lignin alfalfa. Forage quality testing is currently underway at Cornell. In conclusion, although low lignin alfalfa showed comparative yield to standard types in pure stands, it may be less competitive when planted in binary mixtures with cool season grasses.

Is there an association between sex ratio and the level of sex expression among plant populations? A study of Plagiochila porelloides (Torr. ex Nees) Lindenb.
Sexual reproduction promotes genetic diversity and population persistence, relying on presence of both sexes. Variation in the numbers of males and females, including differences in sexual development, result in sex ratio (SR) variation. In fact, in many plant species one sex can be rare or absent, negatively impacting sexual reproduction. Many studies on SR have recognized the potential impact of non-reproductive individuals on population dynamics, but exclude these individuals for the obvious reason that they do not contribute to the current population SR. Further the explicit relationship between sex expression (SE) level and SR is rarely considered. Here we suggest an intuitive relationship between the SE level and SR of a population. Generally, males in many plant species begin development of sex structures before females. Thus, SR should be initially male bias and then shift to more female bias as more plants produce sex structures. Thus, we predict that the SR (proportion of females) will be positively associated with the level of population sex expression (proportion of population producing sex structures). Sixteen populations of Plagiochila porelloides from Robinson Forest field station (University of Kentucky) were surveyed for SE level and SR to test the proposed relationship. The observed the SE level varied from 0.02 to 0.49 and SR varied from 0.01 to 0.85. There was no significant relationship between SR and SE ratio. This result might be due to different total population SR and SE level among populations, and/or different environmental conditions affecting the onset of sex structure development.

Morus rubra (Red Mulberry) is a native tree in the forests of eastern North America; M. alba (White Mulberry) is an invasive, non-native weedy species, most common in disturbed places. Both species are wind-pollinated and they readily hybridize. Frequently, interspecific hybrids predominately backcross into just one of the original parent species. This results in cryptic hybrids, which display few recognizable traits from one of the original parent species. Previous work in this lab has identified nuclear and chloroplast molecular markers to distinguish the two species. Restriction sites were identified in the species-specific sequences, which were used to detect individuals with hybrid ancestry. These analyses were compared to voucher herbarium specimens and original field notes to identify field characters indicative of hybrid origins.

Is wintercreeper's (Euonymus fortunei) success as an invader due to avoidance of herbivory?
Nellie S. Heitzman and Sarah R. Bray
Department of Biology and Environmental Studies, Transylvania University,
Lexington, KY 40508

Wintercreeper (Euonymous fortunei) is an evergreen vine from East Asia that has become invasive in North America. Invasive species have been hypothesized to be successful because they are released from enemies such as herbivores and can invest less in defense. We hypothesized that E. fortunei experiences lower herbivory rates than functionally equivalent native vines Parthenocissus quinquefolia and Vitis vulpina as well as hosting a unique suite of insects compared to native vines. Using beat-sheets and sweep-netting on three dates in the summer of 2017, we examined insect abundance of 5 climbing vines of the three species as well as the ground-creeping vines of E. fortunei, On each sampling date, we also estimated the percent leaf area lost to herbivory on 20 leaves per species. Herbivory varied among species with ground E. fortunei experiencing the lowest herbivory rate and P. quinquefolia the highest (H=110.19, p < 0.001). We found a difference in insect diversity as measured by Shannon-Weiner index among vines with ground E. fortunei having the highest diversity (F3,56 = 3.5, p = 0.02). There was no difference in insect abundance among vines (F3,56 = 1.908, p = 0.14). Our results suggest that E. fortunei may be escaping herbivory by a lack of natural predators in the region- supporting the enemy release hypothesis. The increased insect diversity found in ground E. fortunei may suggest that, while it does not provide food for native insects, it may alter insect community composition via a change in the structure of the forest floor.

The landscape of many college/university campuses serves as both a place of beauty and contemplation, as well as a botanical resource. The focus of this project was to create an online website that would provide students, staff, and faculty of Bellarmine University (Louisville, KY) with the location and identification of 30 different plant species growing on campus. While researching online tree maps from other college campuses, it was observed that many lack detailed photographs that clearly show identifying characteristics of the plants. Therefore, we took numerous photographs and created collages composed of both landscape and close-up images, including distinctive features such as bark texture, leaf arrangement, and leaf shape. The interactive mapping software (provided by ESRI ArcGIS) is a program that is free, accessible, and relatively easy to use. The online map includes a zoomable satellite image of the campus with the 30 different trees or shrubs indicated by numbered markers located in the correct position on the map. This interactive map is published on the internet (http://arcg.is/2bKw7Rm), thus providing the entire Bellarmine community with a resource for understanding more about the flora surrounding them. It should also be useful for students studying botany or ecology. We recommend other college campuses attempt the same thing.

The Drosophila Air Sac Primordia (ASP) can be used to study invasive behavior as it mimics tumor behavior when it invades into the Wing Imaginal Disc (WID) during larval development. A protein expressed in the ASP was found to be coded by the gene pebbled (peb). This study was conducted to see whether pebbled influences the invasive behavior of the ASP. Using Drosophila genetics, the pebbled gene was knocked down and less-developed and less-invaded ASP's were seen as the result. The data presented provides information on the possible role of pebbled in tumor invasion. The downregulation of pebbled also lead to a decrease in filopodia (actin projections protruding from the ASP) supporting the claim that there is a direct positive correlation between quantity of filopodia and the invasiveness of the ASP.

Abnormal development of the vertebrate eye can result in long-term defects, one of which is ocular coloboma. Ocular coloboma is a congenital malformation that occurs when the choroid fissure fails to close during embryonic development. It can result in visual impairment and is also associated with other morphological defects like micropthalmia and cataracts. SOX4 is a transcription factor that has been shown to be associated with ocular coloboma. SOX4 belongs to the SoxC subfamily of proteins, characterized by the presence of a high mobility group (HMG) domain-containing Sry-box, that allows the proteins to bind to DNA, and a transactivation domain that allows the proteins to function as a transcription activator. In previous studies, morpholinos were used to knock down the expression of Sox4 in zebrafish. This resulted in coloboma, micropthalmia, and fewer rod photoreceptors. These morphant phenotypes were supported by preliminary data observed from individual CRISPR genetic mutants in sox4a and sox4b. However, the mutants have a less severe phenotype than the morphants and previous attempts to make a double mutant were not successful. To determine whether mutation of one sox4 co-orthologue sensitizes the zebrafish to loss of the second co-orthologue, we injected a low dose of sox4b morpholino into sox4a mutant embryos, and compared the penetrance and severity of the resulting ocular phenotypes to control injected and single mutant embryos. The data presented here suggest that a low dose of sox4b morpholino does increase the penetrance and severity of some of the microphthalmia phenotype.

CP1 is a well-conserved cathepsin L-like protease essential for proper growth and development in Drosophila melanogaster. Previous research has demonstrated that CP1 has the ability to break down the extracellular matrix. Using the UAS-GAL4 system, immunohistochemistry, and antibody-staining, this research attempts to characterize the role of CP1 and its effects on basement membrane degradation and signaling. These effects include actions at the cellular level and on a known signaling pathway. The genes involved in this pathway are known to be required for proper development of the wing disc into the adult wing. These genes also have human homologues that play important roles in human development. Results from our knockdown experiments will be presented.
Understanding how CP1 affects Drosophila development through cellular and gene activity is important because cathepsins are highly conserved between flies, humans, and have been implicated in several diseases, including cancer. Discovering the mechanisms by which CP1 functions allows for discoveries to be made in connection with disease processes.

During early stages of embryonic development, cells must undergo differentiation to form various tissues and organs. Cells express various genes as they undergo differentiation, but not all of these genes are known. We have identified several genes that may play an important role during development. Minichromosome maintenance complex 3 (MCM3) is important in DNA synthesis. Palmdelphin (PALMD) is involved in cell membrane structure. Phosphoenolpyruvate carboxykinase 1 (PCK1) is an enzyme that metabolizes glucose. To observe mcm3, palmd, and pck1 during development, we fixed Xenopus embryos and used in situ hybridization to determine the expression patterns of these genes. We can also verify gene expression at various embryonic stages with Polymerase Chain Reaction (RT-PCR). We found that each gene has a unique expression pattern during embryonic development. In the future, it will be interesting to investigate what roles MCM3, PCK1, and PALMD might play during the differentiation of the vertebrate embryo.

Individualization is the evolutionarily conserved process during which interconnected spermatids are separated from each other by actin-rich structures called individualization complexes. Although the genetics behind it are poorly understood, recent studies in Drosophila have implicated the JAK-STAT signaling pathway. JAK-STAT is a highly pleiotropic and evolutionarily conserved pathway with roles in processes including reproduction, immunity, cellular differentiation, apoptosis, and carcinogenesis. In testes, activity of the pathway is required in cyst cells, which are somatic support cells similar to mammalian Sertoli cells, for initiation of individualization. It is hypothesized that JAK-STAT activates genes in cyst cells which send an unknown signal or signals to germline cells, initiating individualization. The goal of this project was to identify these genes. To accomplish this, we used GAL4-driven RNA interference to impede the expression of 14 candidate genes in the cyst cells of fruit flies. These 14 genes are all known to code for signaling molecules, and exhibit reduced expression in JAK-STAT impaired testes. We then tagged actin with phalloidin, allowing us to quantify the individualization complexes in each knockdown line. We found that impeding the expression of most of our candidate genes significantly impaired individualization, and that impairing some of these genes in germline cells also impaired individualization. These results suggest that most of these genes may play a role in signaling germline cells to initiate individualization in Drosophila.

Inclusion body myopathy (IBM) is the most common myopathy in people over the age of 50. It causes slow progressive muscle weakness and is characterized by vacuolization, mitochondrial dysfunction, endomysial inflammation, and cytoplasmic deposition of TDP-43.
Recently, whole exome sequencing has recently shown that valosin-containing protein (VCP) is a significant risk factor in sporadic forms of IBM. In addition, mutations in VCP are known to cause a hereditary form of IBM associated with Paget's disease and frontotemporal dementia (IBMPFD). To better understand the disease pathology of IBM, the Lloyd lab expresses a mutant form of this gene in Drosophila, which recapitulates human IBM pathology. Importantly, this model shows TDP-43 depositions in the cytoplasm, and it has been shown that such aggregations are sufficient to interfere with nuclear cytoplasmic transport (NCT). As disruption of NCT is a key pathologic event in other TDP-43 proteinopathies, such as ALS, the purpose of this study was to identify if IBM pathology was driven by aberrant NCT. A genetic screen using Bloomington Drosophila lines and a mutant VCP line was performed, as well as subsequent climbing assays and thorax dissections. Two Bloomington lines contained genes that enhanced the IBM phenotype; future investigation into the NCT genes of each line may shed light on their role in IBM pathogenesis.

The pharynx is a region in vertebrate embryos that gives rise to multiple organs including the cartilage of the head and neck, thymus and parathyroid glands. If the pharynx does not develop properly, it can cause birth defects such as DiGeorge Syndrome. We hypothesized that the FGF signaling pathway plays an important role in the development of the Xenopus pharynx as in other vertebrates. We manipulated FGF signaling in Xenopus embryos at various stages of development. In situ hybridization of control and FGF manipulated embryos showed a change of RA pathway gene expression in manipulated embryos. These results suggest that FGF signaling plays a key role in Xenopus pharyngeal development at least partially by regulating RA signaling. Future work will determine the exact interactions of the FGF and RA signaling pathways. These results will aid in our understanding of the genetic cues that guide vertebrate pharyngeal development and how disruptions in these cues may result in birth defects.

Benzo[a]pyrene (B[a]P) is a carcinogenic pollutant found in cigarette smoke, vehicle exhaust and grilled food. Recent studies monitoring pregnant women and their children revealed that high exposure to polycyclic aromatic hydrocarbons such as B[a]P affect the developing brain and lower IQ at school age. Our previous studies with polychlorinated biphenyls found that genetic differences increase susceptibility to exposure during pregnancy and breast-feeding. We use a mouse model to mimic human genetic variation and followed up our previous experiments by looking at motor function and reflexes in mice during the first two weeks of life. Two tests were administered, surface righting reflex and negative geotaxis, from postnatal day 5 (PND 5) through PND 14. We hypothesized that pups exposed B[a]P during early brain development would show impaired function on both tests. We found significant differences in the righting reflex test on P7 and in negative geotaxis on P10 and P14. Surprisingly, B[a]P exposed mice had longer latencies in negative geotaxis on P10, but significantly shorter latencies on P14. Together, these results suggest that early life exposure to B[a]P causes a delay in the development of normal motor function.

Autophagy is a cellular degradation system that delivers cytoplasmic elements to the lysosome. This process involves various factors known as autophagy-related proteins. One of them, Vps34, is critical to the early stages of autophagy and also contributes to processes such as endocytosis and phagocytosis. Studies have shown that Vps34 plays a survival role in many important cellular types, and that global depletion of Vps34 causes autophagy dysfunction and cell death. Therefore, analysis of Vps34 function in distinct tissues requires tissue- or cell type-specific Vps34 gene deletion, a process called conditional gene-deficiency. The Van Kaer and Wu labs have generated mice selectively deficient in Vps34 only in adipocytes to understand the role of Vps34 function in metabolic disease. Studies with these animals showed that the animals develop insulin-resistance, which is a precursor for the development of diabetes. Herein, we analyzed stained tissue sections of liver, spleen, and adipose tissue of mice with and without the Vps34 gene in adipocytes. The inflammatory status and immune cell composition within adipose tissues were also analyzed.

Current treatments for fibrotic diseases include modulating the immune response by inhibiting macrophage function. While over-active macrophages can lead to increased production of collagen and other scar-forming molecules, research has shown that these cells are also essential for tissue regeneration. In this study, we use a recently developed model of mammalian tissue regeneration, the African spiny mouse (Acomys cahirinus) to investigate macrophage activity during regeneration. We hypothesized that unique macrophage activity in Acomys promotes regeneration over scar formation. To test this hypothesis, we isolated bone marrow derived macrophages (BMDM) and stimulated macrophage maturation to an M1 (classically activated) or M2 (alternatively activated) phenotype in vitro. Previous studies in Mus have demonstrated that M1 macrophages produce high levels of Cd86 and Socs3 whereas M2 macrophages produce high levels of Cd206 and Tgf1. Using these macrophages from both species we quantified phenotypic differences in gene expression. First, we observed that Acomys failed to upregulate Cd86 across all phenotypes, suggesting this Mus M1 markers is not conserved across species. Instead, we found that Socs3 was strongly upregulated after M1 stimulation supporting this as a cross-species marker for classical stimulation. Next, we observed that macrophages from both species responded similarly to M2 stimulation based on upregulation of Cd206 and Tgf1. Lastly, we found that macrophages from both species express collagen 1 and low levels of fibronectin. Together, our results suggest that macrophages from different species can respond differently to specific external cues and may directly contribute to matrix production.

Unlike most vertebrates, sea lamprey embryos undergo unique changes that re-engineer the physical structure of their genome. These restructuring events (known as programmed genome rearrangements) result in the selective removal of several hundred genes from essentially all somatic cell lineages: they are only retained by the germline. Previous studies have found that this loss of DNA is highly orchestrated and presumably involves changes in the movement of eliminated DNA during anaphase. However, our understanding of the processes of DNA elimination has been reconstructed from snapshots of fixed embryos and the process has never been observed in living embryos. My work seeks to optimize DNA staining, imaging and embryo culture methods that permit the real time visualization of DNA elimination in live, unfixed embryos.

Approximately 600,000 US citizens die from cardiovascular disease annually, including males and females across all ethnic groups. The development of diagnostics and treatments often ignore the differences in biochemistry and genetics involved in disease development and presentation. Our objective is to define the molecular basis for sex-specific pathophysiologic differences through the study of differential gene expression in heart failure. Using RNA extracted from myocardial tissue and Taqman PCR to measure expression levels of both mRNA and miRNA (inhibitory RNA, which do not become proteins), we have defined sex specific patterns of genes involved in cardiac fibrosis and contractility.

Mycobacteriophages are viruses that infect bacterium in the genus Mycobacterium and are readily found in environmental samples. The Science Education Alliance-Phage Hunters Advancing Genomics and Evolutionary Science (SEA-PHAGES) is a mycobacteriophage research project that has two distinct portions: phage isolation and phage genome annotation. In the spring of 2016, two mycobacteriophages were discovered from various soil samples in North Central Kentucky using Mycobacterium smegmatis as host. These two mycobacteriophages, Spock and Inky, were difficult to amplify and titer but were sufficiently amplified for DNA extraction at Western Kentucky University. Their genomes were sequenced at North Carolina State University and both Spock and Inky were determined to belong to subcluster K1. These genomes are being annotated this fall using the Phage Evidence Collection and Annotation Network (PECAAN). Phages in subcluster K1 typically have a 11 bp 3' overhang. Preliminary results suggest that the Spock and Inky genomes encodes 96 and 95 gene respectively and no encoded tRNA. The K1 subcluster is of particular note because most of the mycobacteriophages in this cluster can infect Mycobacterium tuberculosis . This research will add to the growing knowledge of mycobacteriophages, with the ultimate goal of discovering new lab techniques and treatments for the pathogen M. tuberculosis.

Direct detection of double-stranded DNA of Staphylococcus aureus using engineered zinc finger proteins immobilized on paramagnetic beads

The facile and specific DNA detection methods are in great need for clinical (i.e., point-of-care (POC) testing) and biomedical applications. The detection system of low volume of pathogen-specific DNA sequence has been demonstrated by immobilizing a fluorescence conjugated zinc finger proteins (ZFPs) on small paramagnetic beads (2-3 micrometer in diameter). The small diameter beads provide a larger surface area to volume ratio, which would help to detect more target DNA due to the increasing amount of ZFPs immobilized on the beads. The ZFP-based detection system allows for direct detection of pathogen-specific double-stranded DNA (dsDNA) without target DNA amplification. Here, ZFPs are engineered to detect staphylococcal enterotoxin B (SEB) which is released by Staphylococcus aureus. The fluorescence-based assay has been applied to monitor immobilized ZFPs labeled with Alexa 488 and the bound complex of ZFPs and Cy5-labeled pathogenic dsDNA. The ZFP-based detection of pathogenic dsDNA has demonstrated a sufficient sensitivity, suggesting a potential to be developed into a simple and reliable method for pathogen detection.

Post-transcriptional modifications are abundant in tRNA and are required for proper function. 2'-O-methylation of tRNA residues 32 (Nm32) and 34 (Nm34) is conserved among eukaryotes, including yeast and humans. In yeast, the Trm7 methyltransferase interacts with Trm732 and Trm734 to form Cm32 and Gm34, respectively, on tRNAPhe. Human FTSJ1, the homolog of Trm7, and human THADA, the homolog of Trm732, interact to form Cm32. Mutations in FTSJ1 cause intellectual disability and mutations in THADA have been linked to polycystic ovary syndrome and type 2 diabetes. We are currently studying Trm732 in two ways. First, to determine the function of Trm732 in the Trm7 methyltransferase reaction, we are generating protein variants and testing their function in yeast using a growth assay. The Cm32 levels on tRNA from yeast expressing these variants will also be quantified to determine whether they correlate with cell growth. Second, to determine the biological role of Cm32 and Gm34 on tRNAPhe, we are studying genetic interactions by testing the growth of yeast trm732Δ and trm734Δ double mutants. We find that the genetic interaction between TRM7 with HDA1 and SRC1 is likely due to lack of Gm34, because hda1Δtrm734Δ and src1Δtrm734Δ yeast mutants are sick. We also find that PUB1 interacts with TRM732, but not TRM734. This finding is consistent with high throughput studies, and indicates that Cm32, but not Gm34 is important for growth in pub1Δ mutants. We are currently working to identify the hypomodified tRNA(s) responsible for the growth defect in each of these strains.

The nucleolus is a dense area in the nucleus when a cell is in interphase. One of the main functions of the nucleolus is ribosomal biogenesis. Malfunctions or dysfunction in any step of the production of ribosomes can induce ribosomal stress. Ribosomal stress effects many proteins and can lead to main diseases. In our research, we are directly focused on how ribosomal stress effects ribosomal protein (RP) L11 which is an important component of the large ribosomal subunit that loads 5S rRNA. RPL11 also plays a significant role in regulating growth associated pathways. Ribosomal stress causes RPL11 and ubiquitin ligase MDM2 to interact and cause tumor suppressor p53 activation leading to apoptosis. Through bio computer screening, thirty-five drugs were selected that targeted L11. The drugs selected had the potential to disrupt and promote ribosomal biogenesis. Through testing, it was found that the drugs were quite toxic more than aid in cell survival. Therefore, the top nine cytotoxic drugs showed potential to be used to aid in anticancer therapies and were chosen to be tested on other cancer lines. Afterwards, the drugs' effect on nuclear morphology was analyzed.

Discoidin domain receptor 1 (DDR1) is a receptor tyrosine kinase that binds to and is activated by collagens. Although activation of DDR1 is required for normal tissue development, DDR1 upregulation and/or activation following injury is detrimental in conditions such as cancer, atherosclerosis, and fibrotic diseases. The focus of our research is to determine the role of DDR1 in kidney fibrosis. We showed that loss of DDR1 reduces fibrosis and improves renal function in models of kidney disease. In addition, cells expressing kinase dead DDR1 produce significantly less collagen than cells expressing wild type DDR1. This result indicates that the kinase activity is required for DDR1-mediated pro-fibrotic effect. Based on this finding, our goal is to develop a small molecule ATP-competitive inhibitor that can selectively inhibit the kinase activity of DDR1. Using structure-activity relationship optimization and time-resolved fluorescence energy transfer assays, we synthesized and screened 95 derivatives of a previously characterized inhibitor, Compound 1 (IC50 for DRR1 = 11.9nM), in order to generate inhibitors with better selectivity and lower IC50. Because of the high homology between DDR1 and DDR2, we tested potential inhibitors for selective inhibition of DDR1 versus DDR2. Our results revealed 14 promising inhibitors, three of which showed a lower IC50 than Compound 1. However, all 14 small molecules inhibited both DDR1 and DDR2. We are in the process of refining the structures of the 14 compounds to improve selectivity and specificity. Our ultimate goal is to use these inhibitors in the setting of fibrotic diseases.

Research was supported by a Veteran's Affairs Merit Awards 1I01BX002025. This work was also supported by the Aspirnaut Program through the National Institute of Diabetes and Digestive and Kidney Diseases of the National Institutes of Health, R25 DK09699 to B.G.H, Vanderbilt University Medical Center.

NADPH oxidase is a multi-subunit enzyme that generates superoxide, which is converted to antimicrobial reactive oxygen species(ROS). Mutations in NADPH oxidase subunit alleles that ablate enzyme function cause chronic granulomatous disease(CGD), an immunodeficiency with life-threatening bacterial or fungal infections, and granulomatous inflammatory complications in the gastro-intestinal and respiratory tracts unrelated to infection. Separately, ROS has an independent, essential role in immune regulation. Emerging evidence links hypomorphic NADPH oxidase alleles with enhanced susceptibility for multiple chronic inflammatory and autoimmune disorders. Mechanistic understanding of how NADPH oxidase-derived ROS regulate inflammation is largely lacking. We determined whether NADPH oxidase regulates inflammatory responses downstream of Toll like receptor(TLR) stimulation. Bone marrow-derived dendritic cells (BMDCs) from wild type(WT) or Cybb-/- mice that lack NADPH oxidase function due to deletion of Cybb were stimulated with TLR2 agonist Pam3CSK4. Inflammatory cytokine expression was determined by quantitative PCR and ELISA. BMDCs from Cybb-/- and WT mice had similar surface expression of TLR2 and co-stimulatory molecules CD80, MHC-I, and MHC-II. Pam3CSK4 induced TLR2 stimulation strongly activated NADPH oxidase resulting in ROS production in WT BMDCs measured by lucigenin-elicited chemiluminescence. Q-PCR analysis showed similar levels of pro-inflammatory cytokine transcripts (TNF-α,IL-1b, and IL-6) on TLR2 stimulation within the two genotypes, indicating ROS production does not influence transcriptional responses in BMDCs. However, Cybb-/- BMDCs expressed elevated protein levels of IL-6 and TNF-α(ELISA) compared to WT BMDCs. These data demonstrate hyper-inflammatory responses in Cybb-/-BMDCs are partly driven by overt production of pro-inflammatory cytokines. Future studies will determine redox-regulated signaling proteins and pathways activated by TLR2 that are regulated by NADPH oxidase derived-ROS. Overall our preliminary studies indicate a counterintuitive role for ROS modulating host inflammatory responses. These studies are essential for understanding the basis of CGD hyper-inflammation.

Periodontitis is a poly-microbial disease characterized by chronic inflammation of the gums and the eventual destruction of alveolar bone. The progression of gum disease into periodontitis is partially driven by a shift in the composition of the oral microbiota from symbiotic to a dysbiotic, more complex community that can evade killing while promoting inflammation. Neutrophils are white blood cells that act as the innate immune system's first responders to infection. They play an important role in periodontitis because they are the main phagocytic cells in the periodontal pocket. During homeostasis, they can eliminate bacteria through the generation of reactive oxygen species and the microbicidal contents of their granules, among other mechanisms. However, oral pathogens have evolved defenses against neutrophil killing and further promote inflammation to harvest nutrients for bacterial growth. The newly appreciated bacteria Filifactor alocis, has been found in lesions of periodontitis patients, but not in the oral microbiota of healthy individuals. F. alocis is strongly resistant to killing and can manipulate neutrophil (PMN) effector functions. Suppressor of Cytokine Signaling 1 (SOCS1), is a protein that acts as a negative regulator of signaling. In PMNs subject to an F. alocis challenge, SOCS 1 mRNA is increased dramatically. RNA-sequencing results found the SOCS 1 mRNA goes up by twelve-fold after six hours in comparison to untreated cells. The RNA-seq was verified by qPCR, providing similar results. Confocal imaging and SOCS1 immunostaining was used to find intensity of fluorescence of the SOCS1 protein in F.a challenged cells. F.a was tested at 1hr, 3hr, and 6hr time points, along with a negative control of untreated cells and a positive control of cells stained with LPS or FSL-1. The results indicate SOCS 1 is increased in F.a challenged PMNs. This shows F.a likely can manipulate neutrophil function by affecting the SOCS1 protein.

The lack of regulation and overuse of growth promoting antibiotics like Tetracycline in agricultural feeds leads to significant amounts of antibiotics in livestock waste. These excreted antibiotics allow for the continuous development of antibiotic resistance genes in the environment. Antibiotic resistance is becoming an increasing risk to public health. Subsequently, antibiotic resistance deaths are projected to surpass cancer related deaths by 2050. The goal of this study is to determine the abundance and persistence of tetracycline (tet) resistance genes in swine waste over a period of 100 days in an anaerobic digester system. Tet(A), tet(B), tet(G), tet(M), tet(O), tet(Q), and tet(W) were quantified by quantitative polymerase chain reaction after DNA extraction. Primers that target ribosomal protection proteins and efflux proteins were used. Antibiotic resistance genes decreased from day one, but were found to be present throughout the study.

tRNAs are molecules that translate mRNA into proteins. tRNAs must receive chemical modifications for proper function, and incorrect modifications can lead to defects such as cancer. The 3-(3-amino-3-carboxypropyl) uridine (acp3U) modification is formed on some bacterial, plant, and animal tRNAs by an unknown enzyme.  To identify this enzyme, primer extension of tRNA from E. coli strains lacking genes encoding predicted methyltransferases with no known function were tested for the presence of acp3U, which causes a primer extension block. Eleven candidate genes have been tested, and none appear to be responsible for the modification. In a complementary approach to identify the enzyme, acp3U formation activity is being purified from E. coli using various protein separation techniques. To detect enzyme activity, a radiolabeled tRNA is incubated with protein, digested, and analyzed by thin layer chromatography to detect modified nucleotides. Identification of the bacterial enzyme will allow study of the function of acp3U.

The TOR (Target of Rapamycin) pathway is a central regulator of cellular growth and is highly conserved from yeast to humans. Our lab has previously identified mutations in S. cerevisiae genes in the TOR pathway that result in genome instability. This project focuses on eight genes of interest that are involved in regulation of TOR signaling as part of the EGO/GSE complex. The EGO/GSE complex is responsible for nutrient sensing and regulation of microautophagy via TOR signaling. These genes include EGO1, EGO3, GTR1, GTR2, TCO89, CDC60, VAM6, and LTV1. Heterozygous, along with most homozygous, knockout strains for these genes have been created and assayed for genomic instability. The two assays used to assess genomic instability are measurement of loss of an exogenous chromosome fragment (via a sectoring assay) and measurement of the loss of the endogenous CAN1 allele (via a fluctuation analysis assay). The EGO1 and CDC60 heterozygous knockouts were found to significantly increase genomic instability according to our fluctuation analysis assay. This may indicate that both EGO1 and CDC60 are dosage sensitive mediators of genomic stability.

We use a program called AMBER, on Western Kentucky University's computing cluster, to run molecular simulations. AMBER uses a Linux command line and is good at organizing data and files. For each protein that we ran using AMBER we created a new directory to organize the files that go with each individual protein both left and right. We used a basic min, heat, and prod file for each protein that we ran and a submit file to go along with each of the min, heat, and prod files.

To get the information about the individual proteins, we download the pdf of the proteins and then we run tleap to create the prmtop, a list of all the atoms in the protein, and inpcrd, coordinates for the location of the individual atoms in the complex protein, files. We then run the submit files to run the protein. We heat the proteins to a very high temperature and then cool the proteins back down to see if they have changed their confirmation. Once all the files have been submitted and have finished running, the files can be viewed by importing files to VMD (visual molecular dynamics). By importing the inprcd files it shows how the protein changes throughout the running of the simulation, therefore can be used to predict a proteins conformation. Also, RMSD (root-mean-squared deviation) analysis is used to measure the deviation of a reference set of coordinates to a target set of coordinates.

Eventually we hope to be able to compare the bound and unbound proteins, extract interesting structures from our MD runs, and perform tagdock on our proteins.

When MDA-MB-231 cells are injected into the mammary fat pads of mice, they either remain at the location of injection as TMD cells or metastasize to the bone as BMD cells. Recent research has demonstrated that there are identifiable differences in the genetic codes of pre-metastatic, migratory TMD and metastatic, non-migratory BMD breast cancer cells. Specifically, it has been shown that the S100A4 and GRM3 genes are downregulated after metastasis. Suppressing these genes may lead to the prevention of bone metastasis. Current methods of observing these differences between BMD and TMD cell lines include nucleic acid sequence analysis, which is invasive and necessitates harvesting cells. Numerous studies have demonstrated the use of Solid Phase Microextraction (SPME) and Gas Chromatography-Mass Spectrometry (GC-MS) to analyze Volatile Organic Compounds (VOCs) in the headspace of cancer cells. SPME concentrates VOCs in the headspace onto a fiber and GC-MS separates and identifies the compounds. For this project, it was hypothesized that there are differences in the VOCs present in the headspace of TMD and BMD breast cancer cells. The determination of these differences could lead to less invasive ways of differentiating between BMD and TMD cancer cells. To identify the VOCs in question, a SPME fiber was placed in the headspace of flasks containing BMD or TMD cells to adsorb the VOCs and injected into the GC-MS instrument for analysis. Results were compared statistically to identify measurable differences in VOC signature. Initial results demonstrate that there are differences in the VOC profiles of the two cell lines and SPME/GC-MS is able to noninvasively differentiate between TMD and BMD breast cancer cells.

Tetracycline antibiotics are widely used in agriculture to prevent infections in livestock and to promote animal growth. Large amounts of antibiotics are released into the environment through animal waste, which can subsequently lead to the spread of antibiotic resistant bacteria. Anaerobic digestion is one of the methods used to reduce the concentrations of antibiotics and antibiotic resistant bacteria.
In this study, tetracycline antibiotics (tetracycline, oxytetracycline, and chlortetracycline) are spiked onto cattle, swine, and poultry manure and the concentrations of the tetracyclines and their main metabolites are monitored over a 64 day period. Air-tight PVC tubes are used during the anaerobic digestion process, and samples are taken every 8 days. The tetracyclines in the manure are extracted using a mixture of methanol and a sodium-EDTA buffer. The tetracyclines were trapped on polymeric weak cation cartridges and analyzed using Liquid Chromatography-Mass Spectrometry (LC-MS). The anaerobic degradation rates of the tetracyclines used in this study will be presented.

Drug abuse is considered a national epidemic in many countries including the USA and is associated with impaired socio-economic loss and adverse public health. Wastewater based epidemiology (WBE) is a cost-effective, comprehensive, and a non-invasive technique capable of determining semi-real-time community usage of drugs utilizing the concentration of drug residues in wastewater, wastewater inflow, and the population served by the centralized wastewater treatment plant. An analytical method capable of simultaneous analysis of 10 illicit drugs, 19 prescribed drugs, and their 7 metabolites was developed and validated. Wastewater influent, effluent, immediate receiving water (Bee Creek), and the Clarks River water were sampled for seven consecutive days in Murray, KY. Samples were extracted and being analyzed using liquid chromatography-triple quadrupole tandem mass spectrometer (LC‒MS/MS). The limit of quantitation of drug residues were 10 ng/L to 400 ng/L. Triplicate spiking recoveries of drug residues in wastewater and river water ranged from 58±8.3% (oxycodone) to 108±7.2% (venlafaxine).

Hydrogen sulfide (H2S) is a toxic gas with the odor of rotten eggs and is mainly known to
occur as a natural gas. In addition, H2S is hypothesized to be the third endogenous gaseous

transmitter after carbon monoxide (CO) and nitric oxide (NO), with many unexplored sig-
naling functions in the human body. Because of its many unknown roles in the body, the

creation of sensitive H2S probes is beneficial in order to explore H2S functions. Based on
our prior creation of a turn-off fluorescence Schiff base sensor for the sensitive detection
of cyanide (CN–

), similar probes were synthesized by reacting Disperse Orange 3 and 4-
Nitrobenzaldehyde to form one probe and 1,5-Diaminonapthalene and 4-Nitrobenzaldehyde
to form a second probe. Aqueous H2S reduces the nitro groups on both of the probes to
primary amine groups, causing turn-on fluorescence. Both probes were characterized with
1H-NMR, and the process of optimizing solvent ratio, fluorescence excitation wavelength,
and incubation time and temperature was carried out with the first probe. Likely due to
a small Stokes' shift and the interference between the excitation beam and emission, the
fluorescence output of the first probe did not directly correlate to the [H2S]. The process of
optimizing the fluorescence response in both probes is currently in progress.

Polyunsaturated fatty acids (PUFAs) belong to a group of essential nutrients that are necessary for the optimal health of human and animals. Increasing concerns over the sustainable sources of essential PUFAs led to explore the PUFA-rich rations that can enrich ω-3 acids in meat and eggs. The nutritional composition (including linoleic and α-linolenic acids) of industrial hemp products provide opportunities using it as a livestock ration. A mixture of hemp hearts and commercially available soy-based diet (1:1) was fed to an experimental group of twenty-five hens (1 hen/cage) and a commercially available soy-based diet for a control group of twenty-five hens. Egg yolks (a day/week up to six weeks) were composited, freeze dried, extracted, methylated, and being analyzed for twelve PUFAs using gas chromatography mass spectrometry (GC/MS). First eggs were pooled separately and will be analyzed. To our knowledge, this is the first study exploring hemp feed as a potential livestock ration in the USA to enrich PUFAs in chicken eggs.

In a world where electronics are a fundamental part of everyday life, bendable displays with tunable optical gaps are the future of this sector. To determine the validity that halogenated anthracenes and linear aromatic hydrocarbons in general have on the future of this industry, high level quantum mechanical computations were deployed on hundreds of structures of substituted benzene, naphthalene, and anthracene. An in-depth analysis of each halogenated structure shows that the energies of molecular orbitals, specifically the width of the optical gap, is a function of halogens' size, electronegativity, and position on each aromatic ring within a molecule. Interestingly, the lowest optical gap has been found for a structure with the largest molecular polarizability.

Very little research has been reported on the use of instrumental analytical techniques for the forensic examination of contact lenses. However, Chemical and Engineering News projects we will see an increase in the need for corrective eye wear within the next several generations. With the increasing potential of contact lenses being found at crime scenes and their need for analysis, I have devised two methods for analysis of contact lenses using a bench-top FT-IR and an Abbe Refractometer. A bench-top FT-IR identifies the functional groups present in the polymer allowing the analyst to differentiate between different brands. An Abbe Refractometer can be used to find the refractive index (RI) of the contact lens, and therefore hopefully find the prescription of the lens.

Ten contact lenses were analyzed using FT-IR and the spectra obtained distinguished between the different polymers used by each brand. RI 1.332-1.400 were observed for 8 contact lenses, with prescriptions between -3.75 and +3.50 using the Abbe Refractometer. Testing to determine the consistency of these results is ongoing and will be discussed in the presentation.

Contact lenses have the potential to become helpful pieces of evidence in the forensics field and this study seeks to give forensic analysts a procedure for instrumental analysis. FT-IR can distinguish between different polymers which make up contact lenses, and an Abbe Refractometer has the potential to differentiate between prescriptions using the RI.

In pyrophosphate, the J-coupling distribution is strongly correlated with P–O–P bond-angle distribution. By using solid state Nuclear Magnetic Resonance (NMR) J-coupling patterns can be viewed and help determine structural information. The hypothesis that was proposed was that the mixed glasses would have two regions of J-coupling patterns. Rich regions of Pb2+ and another region with both Na+ and Pb2+ patterns. The pyrophosphate samples were synthesized by mixing solid compounds of varying Pb2+ and Na+ ions. The mixed solids was then placed in a muffle furnace and heated to 1100°C. The molten glass was poured onto a copper plate and quenched. The samples were ground to a fine powder and packed into rotors. The samples were analyzed using solid state NMR experiments. PASS (Phase Adjusted Spinning Sidebands) and TE-PIETA (Total-Echo Phase Incremented Echo Train Acquisition). The raw data was then placed into a specialized program to view J-coupling of the PASS and PIETA. Many of the mixed samples displayed as two types of J-coupling. When compared to a pure lead sample the J-coupling was close to 23 Hz; however, the mixed samples seemed to lower the J-coupling to about 14 Hz. We also looked at the chemical shift anisotropy (CSA) of the samples and appears that there is not much variability within them.

Sedimentation Fractionation

8 years' worth of carbon and nitrogen data were acquired and analyzed from sediment traps pulled from the south Elkhorn Creek inlet and outlet stems in Lexington, Kentucky to set a baseline, indirectly representing atmospheric carbon and nitrogen. Because gases in the air are sequestered terrestrially through biological and terrestrial processes, these values can be used as a baseline for comparison to nitrogen/carbon studies in the future. Also, the fractionation process was analyzed over the 8-year duration to support earlier, shorter-term studies on the effects that organic processes have in leaving isotopic residuals. Fractionation occurs because lighter particles are preferred in biological processes since they are easier to digest and incorporate. This leads to an accumulation of heavier isotopic residuals in inorganic sediment.

The 2006-2013 average % value of total carbon is 3.39g / 100 grams of sediment. This value can be used as a starting point comparison as average temperature fluctuates over time. The comparison of the winter months November -February vs. summer growing season March-October shows that the average % carbon 13 ratio was less during winter. This is hypothesized as a result that as less carbon 12 is used for plant growth during winter, increased carbon 12 amounts remaining in the stream would lower the carbon 13 percent ratio.



Reference:

Journal of Hydrology,

Volume 527, August 2015, Pages 908-922

William I. Ford, James F. Fox, Erik Pollock, Harold Rowe, Suvankar Chakraborty

Nanoparticles in dry powder state form agglomerates thus reducing surface energy and accessibility of diamond core impacting technological advancement. In this work, we investigated a facile, cost-effective and contaminant-free salt-assisted ultrasonic de-agglomeration method for detonation nanodiamond, NDs. Utilizing ultrasound energy to break apart two different sourced and thermally treated nanodiamond mesoscale aggregates in sodium chloride and sodium acetate salts, this technique produced aqueous slurry of isolated or single-digit (< 10 nm) stable colloidal dispersions by virtue of ionic interactions and electrostatic destabilization. Moreover, the technique is well-suited for materials engineering (composites, lubricants) and biomedical (bio-labelling, biosensing) applications. We characterized microscopic structure and performed advanced electrochemistry by immobilizing processed NDs on boron-doped diamond to study surface redox chemistry, determine surface potential (or Fermi level), carrier density and to image electrocatalytic activity by scanning electrochemical microscopy and the results are compared to those untreated aggregated nanodiamond particles. The findings are discussed in terms of surface functionality and defect sites that give rise to surface states within bandgap. These surface states may serve as electron donors (or acceptors) depending upon bonding (or antibonding) character suitable for various electrocatalytic redox processes.

Zebra mussels (Dreissena polymorpha) are an exotic and invasive mollusk that spread extensively over various rivers and lakes in the United States. However, these mussels had been unable to reproduce and establish populations in the Kentucky Lake during the last several decades. Recently, zebra mussel colonies were frequently encountered in Kentucky Lake. This emergence of mussels in the waters generated interest in examining physical and chemical parameters that are vital to the reproduction and development of zebra mussels. The specific aim of this study was to examine various physical (DO, pH, light penetration) and chemical parameters (trace metals) in water samples collected from selected long-term monitoring stations in the Kentucky Lake. Surface and bottom water samples were collected during the Kentucky Lake sampling Cruise #572. Pre-cleaned high density polyethylene bottles were used to collect the water samples and filtered using 0.45 micron syringe filters. The filtrate was acidified to 2% nitric acid with trace metal grade nitric acid. Standard analytical procedures were followed including sample preparation, quality assurance protocols and analysis using Inductively Coupled Plasma (ICP) Optical Emission Spectrometer. Arsenic (As), barium (Ba), beryllium (Be), calcium (Ca), cadmium (Cd), chromium (Cr), copper (Cu), nickel (Ni), lead (Pb), selenium (Se) and thallium (Tl) were measured in the samples. Elemental concentrations ranged from µg L-1 (ppb) to mg L-1 (ppm) depending on analyte type and sampling locations. Concentrations of some critical metals were compared with historical (pre-emergence period of zebra mussels) data from Kentucky Lake waters as well as concentrations of these metals in zebra mussel infested Ohio River water samples.

Imidazoles play a vital role in the synthesis of biologically active molecules. These compounds are known to possess diverse biological applications. Benzimidazole derivatives have drawn considerable attention due to their widespread biological applications such as antitumor, antimicrobial, anti-inflammatory, and antihelminthic activities. Various catalysts under different reaction conditions have been employed for the synthesis of these molecules. Most of the reported methods suffer from one or more serious drawbacks such as high reaction temperature, long duration, use of toxic and expensive chemicals as starting materials, use of moisture-sensitive catalysts, and formation of byproducts. In recent years, ZrO2 nanoparticles have gained much attention in catalysis due to their specific amphoteric properties, excellent mechanical strength and stiffness, high thermal stability and dielectric properties. In the present study, we have described the synthesis of imidazoles and 1,2-disubstituted benzimidazoles from readily available benzyl, 1,2-phenylenediamine, aldehydes and ammonium acetate under solvent free conditions using ZrO2-β-CD as an environment friendly and reusable heterogeneous catalyst. The products have been characterized by spectral methods. It is note-worthy to mention that, ZrO2-supported β-cyclodextrin nanoparticles have never been used in the field of synthetic organic chemistry. The nanoparticles (ZrO2-β-CD) prepared by a simple one-pot-coprecipitation method, were characterized by PXRD, SEM, and TEM techniques. The nano (ZrO2-β-CD) particles were found to be an effective heterogeneous reusable catalysts for the synthesis of imidazoles and benzimidazoles under solvent free conditions. Further, the synthesized molecules were evaluated for their antibacterial activity against six bacterial strains. Since the biological results seem promising, further experiments have been designed to understand their mechanism of action.

Clean, available drinking water is an obvious necessity throughout the world. Unfortunately however, many remote and/or impoverished areas are without a reliable potable water resource. Purification systems can be expensive, complicated or both. The need and availability of cheap, reliable methods for purifying water in less developed, small rural areas is unequivocal.
Adsorption studies of organic residues have been reported using magnetic biochar.1 The referenced procedure allows for the preparation of magnetic biochar in the conventional approach of reacting Fe2+/Fe3+ with a NaOH solution to precipitate magnetite onto the biochar surface. Subsequent absorption of organic residues appear to be very effective.
This research seeks to enhance the surface area of magnetic biochar by performing homogeneous precipitation using urea as the source of hydroxide. Preliminary results will be reported on the preparation of biochar and analytical effectiveness in comparison to reported values.
Reference:
1. J.Chem.Educ. 2016, 93, 1935-1938

Junjie Xiang, Yan Cao
Institute for Combustion Science and Environmental Technology, Department of Chemistry, Western Kentucky University, Bowling Green, KY 42101, USA. E-mail: yan.cao@wku.edu

DRIFT (diffuse reflectance infrared Fourier transform) is a powerful tool to provide in-situ IR information of chemical species. Measurement using TG (thermal gravimetric) can provide mass information during the temperature variation under a general chemical reaction condition. Each method spread one critical parameter or property of a chemical reaction along the time scale, enriching reaction information to be extracted. However, it's a great challenge to combine both techniques since both methods required high precision and there is conflict in the basic structure. The main part of TG is a microbalance in a sealed furnace and the main for FTIR is a go through light path. This study explored integration of these two well-know analytical methods for the first time as far as we know. In our design a micro balance is located at the top of the DRIFT chamber. A metal string hangs down hooking the sample pan from the top through the middle of the two parabolic mirrors, one of which focus the IR on the sample while the other collect the IR reflectance for the detector. Underneath the sample tray there is a vertical tube furnace to control the sample temperature. The micro balance have a weight resolution of 0.1 μg , and the furnace can provide sample temperatures ranging from 700 to 980 ℃. This new device can be used to carry out kinetic analysis with more mechanism details.

Heavy metals are used immensely in industrial and consumer products but are unfortunately, easily spread into the environment. By measuring the level of contamination in several samples including: specimens of Aporrectodea complex and the surrounding soil; from a potentially contaminated site such as a junkyard; a risk potential based on ratio of heavy metal in worm to heavy metal in soil was made. The samples were analyzed via ICP-OES to determine heavy metals present. It was hypothesized that these worms would have significantly more heavy metal than the surrounding soil. Because of the accumulation, this chief food source for many animals, could be considered a high risk for biomagnification of heavy metals into the higher trophic levels of the local food chain in the Frankfort area.

CHEMISTRY
Application of metal-organic framework materials in dye-sensitized solar cells. RYAN BOGGESS, BANGBO YAN, Department of Chemistry, Western Kentucky University, Bowling Green, KY 42101.
Dye-sensitized solar cells play an important role in the renewable energy research due to their relatively low production cost, more environmental-friendly manufacture, and mechanical robustness. Recently, our lab has synthesized a series of new metal-organic frameworks (([Ru(H2bpc)M(bpc)(Hbpc)2(H2O)], H2bpc=2,2'-bipyridine-4,4'-dicarboxylic acid, M = Cu, Ni, Fe). Their structures contains zigzag chains of [Ru(bpc)3]n- complex ions linked by transition metal complexes, and shows strong visible light absorption. These framework materials are excellent candidates as dyes in dye-sensitized solar cells because of their visible light absorbing group [Ru(bpc)3]n- and carboxylate groups in the structures. Properties of solar cells made of titanium oxide photoanode and metal-organic dyes will be presented.

P450s are a large family of heme bound monoxygenases that are capable of performing a variety of reactions such as hydroxylation, epoxidation, and carbon-hydrogen activation. These enzymes are well known for their role in human health, specifically steroid synthesis and xenobiotic metabolism. Despite the fact that P450s belong to a large and diverse family of enzymes, they all share a similar fold and utilize the same catalytic cycle. P450 BM3, a bacterial protein that hydroxylates medium to long chain fatty acids, can be used as a model system in order to study the stability of distinct phases of the catalytic cycle. Utilizing site directed mutagenesis we have engineered a BM3 pentuple mutant to act as model system for the more promiscuous human P450s. Biophysical techniques, such as pulse proteolysis and urea titrations monitored by absorbance and circular dichroism, were used. These techniques probe various aspects of the denaturation process, which is supported by results showing their differences in sensitivity to urea. Additionally, using these techniques, we are able to compare selectivity, promiscuity, and determine biophysical differences that arise due to the oxidation state and identity of the substrate or product. Initial results show substantial changes in P450 stability due to differences of oxidation state and substrates/products bound to the protein.

There is an increasing trend in the exploration and discovery of functional luminescent metal-organic frameworks (MOF) because both inorganic and organic can produce luminescence. Moreover, metal–ligand charge transfer transitions within MOFs can induce interesting luminescent functionalities. This presentation focuses on the fluorescence properties of a new metal-organic frameworks (([Ru(H2bpc)Cu(bpc)(Hbpc)2(H2O)], H2bpc=2,2'-bipyridine-4,4'-dicarboxylic acid,). Its fluorescence properties are compared to that of [Ru(H2bpc)3]Cl2. The effects of different solvents on its fluorescence properties are also studied.

Mechanosynthesis, where reactants are ground together to induce reaction, is emerging as an alternative to solution-based, thermally induced methods for producing a wide variety of products and materials. Often accomplished through ball milling, these reactions use little to no solvent and give more sustainable compound preparation through (1) reduced solvent use, (2) reduced reaction time, and (3) reduced waste. The kinetics of the mechanosyntheses of a variety of zinc and palladium dialkyl-2,2′-bipyridyl-4,4′-dicarboxylate complexes will be presented. By fitting the reaction data to a variety of different models and determining how kinetic parameters vary with differences in ligand functionalization and metal salt, guidelines for more efficient preparations of transition metal complexes can be determined. Conclusions on maximizing sustainability in these preparations will be shared.

High-valent metal-oxo intermediates play central roles as active oxidants in enzymatic and synthetic catalytic oxidation. In this work, chemical and photochemical methods are utilized to produce and study high-valent metal-oxo species supported by a 5,10,15,20-tetra(2,6-dichlorophenyl)porphyrin (TDCPP). With iodobenzene diacetate [PhI(OAc)2] as the oxygen source , Iron (III) or manganese (III） porphyrin complexes can convert to the corresponding iron(IV)-oxo and manganese(IV)-oxo species as oxygen atom transfer agents. As an alternative to chemical oxidation, Iron(IV)-oxo porphyrin derivatives were produced by visible light irradiation of the corresponding iron (III) bromate complexes. The photochemical approach produces metal-oxo species essentially instantly, and permits direct detection of metal-oxo species and kinetic studies of their oxidation with a variety of organic substrates.

Cancer research is being conducted due to the alarming death rate of patients. The development of chemotherapy by Rosenberg, has proven to have satisfactory outcomes. However, treatment causes uncontrollable side effects such as nausea, vomiting and hair loss. One strategy to reduce side effects and selectively target tumors is photodynamic therapy (PDT). The Glazer laboratory uses ruthenium compounds as PDT agents because they are less likely to photobleach, exhibit reliable solubility and have tunable properties. One of the tunable properties is the addition of various ligands such as peptides. The goal of this project is to synthesize a peptide ruthenium hybrid that possess anti-cancer activity and can selectively target various membrane proteins. The three essential elements of this project is to synthesize a ruthenium peptide hybrid, characterize the compound and test on cells. The results are ruthenium hybrids that were tested for cytotoxicity in cancer cells and verified cell localization.

N-heterocyclic carbenes have become increasingly popular ligands for organometallic catalysis in recent years. They are found to be a good alternative for more traditional phosphine based ligands. They are fully tunable sterically and to some extent electronically. This research explores a novel synthetic pathway to normal and abnormal carbene complexes. It involves imidazolium based ligands, transmetallation using silver and C-H bond activation with nickel and iridium.

Transesterification of lipids into biodiesel is a promising pathway for the production of renewable fuels. It is synthesized by extracting, purifying, and upgrading the lipids found in various plant and animal feedstocks. Microalgae are a promising source of renewable lipids as these organisms can be grown using carbon dioxide emissions generated by coal-fired power plants. When the lipids are extracted from the microalgae, the proteins and carbohydrates are left behind. These remnants can be used to create bioplastics and bioethanol, respectively. However, the relative amounts of lipids, proteins, and carbohydrates obtained from extraction must be controlled in order for the process to be economically viable on an industrial scale. Therefore, it is necessary to optimize the process for each of the components in order to determine the influence of process parameters on product yields. The experiments conducted here were carried out as part of a design of experiments procedure to determine how each of the process parameters, in this case temperature and solvent volume, would affect the yield of each component.

Photocatalytic reduction of carbon dioxide by metal-organic framework materials. HANNAH BROOKS, BANGBO YAN, Department of Chemistry, Western Kentucky University, Bowling Green, KY 42101.
The design and application of new chemical materials for solar energy harvesting and renewable fuel producing is a very promising and demanding technology because of the diminishing fossil fuel deposits. Our research is to make new photocatalytic metal-organic framework materials consisting of ruthenium polypyridyl complexes and 3d transition metal ions. In this presentation, we will report our study on the reduction of carbon dioxide to formic acid using the new metal-organic framework ([Ru(H2bpc)Cu(bpc)(Hbpc)2(H2O)], H2bpc=2,2'-bipyridine-4,4'-dicarboxylic acid) synthesized in our lab as the photocatalyst. The structures of the catalysts contains zigzag chains of [Ru(bpc)3]n- complex ions linked by copper complex ions, and shows strong visible light absorption. Its photocatalytic reduction of carbon dioxide to formatted and its stability of the material under visible light radiation are studied.

Promotional Effect of Platinum on Supported Nickel Catalysts in the Conversion of Model Lipids to Diesel Range Hydrocarbons:

Given that the high oxygen content of the fatty acid esters that constitute biodiesel causes several problems – including poor cold flow properties, storage instability and engine compatibility issues – that render biodiesel a less than optimal fuel, attention has shifted to the catalytic deoxygenation of the lipids in animal fats and vegetable oils to afford hydrocarbon biofuels. Supported Ni catalysts are active in this reaction, Ni-Cu alloy formulations showing superior performance relative to Ni-only catalysts since Cu improves the reducibility of Ni creating additional Ni0 active sites. In this project, the ability of Pt to promote the performance of Ni-based catalysts in the deoxygenation of a model triglyceride to diesel-like hydrocarbons was investigated. Temperature programmed reduction (TPR) measurements showed that small amounts (0.1-0.5 wt.%) of Pt do facilitate the reduction of Ni at lower temperatures, higher amounts of Ni0 being obtained with increased Pt loading. However, albeit a marked promotion effect was indeed observed in the deoxygenation of tristearin (a model lipid compound) to diesel-like hydrocarbons, the amount of Ni0 obtained did not show a direct correlation with diesel yield, which could be due to decreased interaction between the Pt and Ni atoms at high Pt loadings.

1,3-amino alcohols are a crucial biological and chemical motif and can be used both pharmacologically and in industrial ways. This important functional group can be accessed via the reduction of a Mannich base, a β-amino carbonyl compound, which is formed from readily available starting materials. Traditional methods of reduction include harsh conditions such as high temperatures, the use of strong reductants, or expensive catalysts. This project investigated the reduction of Mannich bases using iron-Knӧlker-type catalysts. This method works for a variety of β-amino carbonyl compounds and utilizes mild conditions: isopropanol as the reductant, and a low reaction temperature (50⁰C).

Boron reagents have been used increasingly, particularly in organic and synthetic chemistry, in recent decades. Tris(pentafluorophenyl)borane has become an important catalyst due to its role as a strong Lewis acid. This catalyst has led to alternative mechanistic pathways in the reduction of a wide variety of functional groups, including the reduction of C-O bonds. As C-O bonds are ubiquitous in nature this has led the desire to develop biofuels and fine chemicals from the cleavage of these bonds. Herein we describe the reduction and cyclization of carbohydrate derivatives, isosorbide and isomannide, using stoichiometric amounts of catecholborane and catalytic amount of B(C6F5)3. Mechanistically, it is hypothesized that the furan rings are reduced selectively at the primary position in the carbohydrate derivatives, generating a 1, 6-deoxytetraol. The tetraol can then form a new furan through intramolecular attack from the C2 to the C5 position. This reduction of isosorbide produces a product with inversion symmetry and three contiguous stereocenters.

Abstract: Antibiotics are some of the most widely known and widely used medications in the world. In fact, in 2009 people in the United States spent some $10.7 billion on antibiotics. Since their discovery, antibiotics have transformed the way bacterial infections are treated. However, due to overuse, the effectiveness of these life-saving drugs is being reduced and antibiotic-resistant infections are becoming a more pressing problem. Oxamazins are a class of heteroatom-activated β-lactam antibiotics that show good activity against a spread of Gram-positive and Gram-negative bacteria. The goal of our research is to progress through a traditional synthesis of a simple oxamazin starting with readily available amino acids before progressing to solid-phase synthesis.

An alternative to storing the energy emitted by the Sun is in covalent bonds of highly strained “chemical fuels” such as bis-anthracene (BA). When BA is exposed to light, highly energetic covalent bonds are formed. We report that simple mechanical grinding with a mortar and a pestle of that mechanophore releases stored chemical energy as heat. We also report a moderate improvement in the synthetic yield of BA. For our future studies on the molecular dynamics and kinetics of the back reaction triggered by pressure or a shear deformation, we are undertaking the synthesis of a selectively deuterated BA and other heteroatom-based analogs in the laboratory.

Metabolic oxidations relating to the protein class known as the cytochrome P450 enzymes hold key knowledge about biological chemistry and the efficient processes undergone to maintain life. These enzymes are present in most metabolic reactions and can function out of body in crucial industrial processes; however, there exist gaping unknowns with respect to them due to their immense nature. In this work, synthesis and characterization of iron and manganese complexes supported by a 5, 10,15, 20-tetrakis-(2,6 dichlorophenyl)porphyrin ligand, abbreviated as MIII(TDCPP)Cl (M = Fe, Mn) was conducted. Oxidation reactions catalyzed by these synthetic catalysts were investigated. As determined by GC-MS spectrometer, remarkable efficiency and high selectivity were observed for a variety of organic substrates. Mechanistic studies were performed to probe for the nature of oxidizing intermediaries relevant to the catalytic reaction. Specifically, the porphyrin-iron(IV)-oxo Compound II models were generated under a known chemical and a novel photochemical method.

CHEMISTRY
Synthesis and properties of a ruthenium polypyridyl complex. HANNAH COPE, BANGBO YAN, Department of Chemistry, Western Kentucky University, Bowling Green, KY 42101.
Ruthenium(II) polypyridyl complexes have wide applications in several research fields such as conversion of solar energy, fabrication of molecular devices, DNA intercalation, and protein binding because their peculiar electrochemical and photophysical properties. In this presentation, we report the synthesis, structure and properties of a ruthenium(II) polypyridyl [Ru(bpy)2(en)]Cl2. Its crystal structure is analyzed using X-ray diffraction. Crystal data: Cc, a=12.7626Å, b=12.9911 Å, c=15.9889 Å, =102.815°, V=2584.93 Å3, R1 = 0.0692.

The design of a hydrophilic heterobifunctional linker with the functionality to conjugate to a monoclonal antibody, mAb, was our research goal. When drug toxins are bound to the linker that is conjugated to a mAb, these molecules are known as antibody drug conjugates, ADC's. Linking drug toxins to mAbs is a way to deliver drug molecules specifically to cancer cells without targeting the healthy cells within the body. This minimizes the toxic effects of chemotherapy treatments. A heterobifunctional linker is functionalized on two separate ends of the small organic molecule. One end of the linker facilitates covalent bonding to the mAb while the other end facilitates covalent bonding to a toxin drug molecule. The mAbs that we are interested in bind specifically to the extracellular matrix (outside cell surface) of cancer cells; in particular antibody CD-55. Our desired linker consisted of a uridine molecule covalently bonded to malemide through an acetal moiety. The importance of the acetal is that this functional group is cleavable at a pH of 5.5 and therefore, once endocytosed (brought inside the cell membrane) within the cancer cell, should cleave to release the drug toxin. Outside the cell membrane is a pH of 7.2. Acetal functions are stable at pH of 7.2 and therefore would remain intact outside the cancer cell. The simulated cancer drug is uridine, however, 5-fluorouridine is a known cytotoxin that can be attached to the heterobifunctional linker in place of uridine once the chemistry has been established. We successfully synthesized the final intermediate precursor to the desired product containing the uridine group. Ideally, once the final product is synthesized, further research would be conducted to demonstrate the cleavage efficiency of the desired compound and then the synthesis of the 5-fluorouridine ADC for evaluation as a cytotoxin specific towards CD-55.

Breast cancer effects one in eight women in their lifetime. According to the American Cancer Society, the overall survival rate for stage IV breast cancer is around 22%. Despite recent advancements, many patients eventually relapse. Cisplatin is a chemotherapeutic drug used for treatment of numerous human cancers including bladder, head and neck, lung, ovarian, and testicular cancers. LHRH is a decapeptide that triggers the production of gonadotropins, which stimulates the growth of eggs in the ovaries and sperm in the testes. The purpose of this research was to synthesize a target-specific anti-cancer agent using cisplatin coupled to LHRH via a malonate linkage that targets the LHRH receptor, which is overexpressed in breast cancer. The results of the synthesis will be discussed in the poster presentation.

Block Copolymers composed of chemical constituents that are incompatible can microphase separate. Similar to the way oil and water are immiscible so they phase separate. Since the blocks are covalently bonded to each other they cannot dissociate in a macroscopic way like oil and water instead they form nano-structures which are governed by the size of each block. Polymer scientists use thermodynamics to describe how the different blocks interact. The product of the degree of polymerization, n, and the Flory-Huggins interaction parameter, chi, gives an indication of how incompatible the two blocks are and whether or not they will microphase separate. For example, a diblock copolymer of symmetric composition will microphase separate if the product chi * N is greater than 10.5. If chi * N is less than 10.5, the blocks will mix and microphase separation is not observed. The incompatibility between the blocks also affects the solution behavior of these copolymers and their adsorption behavior on various surfaces. The structures formed are called morphologies and the most common are lamella cylinder gyroid and body centered cubic. L0 is the distance which when minimized created the most fine nanostructure, Sub 10 nanometer L0 are optimal. Block copolymer self-assembly is generally limited to forming simple patterns such as arrays of dots or lines, determined by the volume fraction of the blocks and the processing conditions. It is desirable to be able to modify both the period of the patterns and their morphology to form more complex structures useful in devices. The period of well-ordered thin films of poly (styreneblock-dimethylsiloxane) (PS-b-PDMS) or poly(styrene-blockmethyl methacrylate) (PS-b-PMMA) can be varied by up to ∼10% by use of an incommensurate template to strain the microdomain array or by changing the annealing conditions.

There is a need for cancer treatment to be exclusively targeted towards cancerous tissue and thus innocuoustowards regular tissue; antibody drug conjugates (ADCs) are a method that utilize a monoclonal antibody (mAb) for targetingcancerous tissues, a drug for its cytotoxic effects, and a small molecule linker to connect the two and specifically release the cytotoxic payload at its intended target. While the theoretical benefits behind them are strong, ADCs lack diverse cleavage methods to deliver their carried drugs. One mechanistic approach for drug delivery would be cleavage of an ADC based on the acidic environment found in cancerous cells (pH = 5.5 in cancerous cells, compared to 7.2 in the systemic circulation). Therefore, theacetal functionality was explored as a novel cleavage mechanism because of its hydrolytic propensity in acidic environments. Two small organic molecule linkers were designed and partially synthesized with the purpose of conjugating chromophores (2,4-dinitrophenyl) to cancer specific mAb to explore the acetal moiety in cellular cancer models, and a non-cleavable small molecule linker was synthesized to develop biological assays for characterizing the designed ADCs. Future studies will evaluate the acetal chemistry for linking cytotoxins in the place of chromophores as cancer therapies.

WeBWorK is an open-source online homework system developed and maintained by the Mathematical Association of America (MAA). This project sought to determine whether WeBWorK would be a useful addition to the mathematics program at Kentucky Wesleyan College and to determine the effectiveness of undergraduate problem authors.

Homework sets for a 100-level mathematics course were created using the Open Problem Library (OPL) as well as through authoring original problems. Editing problems from the OPL and authoring original problems required developing knowledge of Perl, LaTeX, and MathObjects. Further, the author has worked with this course as a supplemental instructor to further evaluate the effectiveness of WeBWorK in the classroom.

During the pilot run in a MATH100 course, which previously did not have any online homework, the students responded well to the addition of WeBWorK. Students enjoyed the immediate feedback on their work and the absence of any license costs to use the software. Further, this project indicated that undergraduate students can be effective problem authors for lower level mathematics courses. Because of the success of this project, the mathematics program is implementing WeBWorK into additional courses. Currently, there are three math classes that are using WeBWorK at Kentucky Wesleyan College with additional courses planned for the coming year.

Today's cutting-edge large scientific research projects are both CPU and data-intensive that require enormous computing power and large-scale data storage capabilities. Grid computing is the new IT infrastructure for the 21st century large-scale Big Data research in science. Grid computing provides the resources that allow researchers to share computer processing power and data across boundaries. At Bellarmine University, we have recently upgraded our Tier3 grid site that is linked to the Open Science Grid (OSG) cyberinfrastructure to a Tier2 grid site dedicated for the LSST (Large Synoptic Survey Telescope) Project. It is currently the only OSG site in the state of Kentucky and the only Tier2 grid site at a predominantly undergraduate institution in the US. The 51-node Tier2 Grid Supercomputer is equipped with 408 cores, 1300GB of RAM and 375TB of disk storage space. OSG Tier2 grid sites have the capability of processing the raw data for the entire collaboration which can also be accessed by others to submit grid jobs via OSG, whereas Tier3 sites typically process a subset of the processed data suitable for analysis, locally. Tier2 grid sites provide a broad range of services to the grid users. We are part of LSST's Dark Energy Science Collaboration (DESC). The Tier2 supercomputer is extensively used to run the Photon Simulator (PhoSim) Monte-Carlo simulation software. We are using High-Throughput Computing (HTC) environment using HTCondor to queue, submit and run jobs to all the nodes using DAGMan that has been configured to run on a Tier2 grid site. We are using Globus to manage, share and transfer large amounts of data with our collaborators via GridFTP. We have deployed Ganglia to monitor the various parameters/metrics such as CPU loads and network utilization of all nodes of the Tier2 Grid Supercomputer, including the inbound and outbound data transfer bandwidth.

Historically, the American Chestnut (Castanea dentata) was an important ecological component of eastern US deciduous forests. Chestnut blight (Cryphonectria parasitica) swept throughout C. dentata's range beginning in the early 1900's and extirpated the tree from much of these areas or made it functionally extinct by the 1950's. Chestnut restoration is a crucial step in reestablishing the natural landscapes and carrying capacities throughout their previous range. Genetically enhanced, blight-resistant chestnuts are being developed, but restoration projects must utilize a spatial aspect to be efficient with initial reintroduction resources. To address this spatial need, we created habitat suitability models for the American Chestnut within Daniel Boone National Forest (DBNF) using both raster and vector overlays in ArcMap. We determined which map layers to include based on their importance to C. dentata as determined by literature reviews on chestnut silviculture and pre-and post-extinction occupancy. The maps will indicate areas that would be most suitable for C. dentata reintroduction and restoration within DBNF. These maps will allow wildlife biologists and conservationists in and around DBNF to make informed decisions when functional American Chestnuts are ready to be reintroduced.

The purpose of this experiment was to quantify and compare detectable elements, including heavy metals, in soil between three different land use practices associated with the Kentucky State University certified organic High Tunnel complex. This was done using the Olympus Delta Premium DP6000-CC XRF with Dual Beam Mining Plus Calibration and Soil Environmental Analysis Package (Waltham, MA), having the capability to measure quantity in parts per million (ppm) of 32 elements, including heavy metals such as aluminum, along with soil nutrients like potassium and phosphorous. On two sampling dates and using ex situ sampling protocol, soil was collected from inside each of the four certified organic high tunnels, from a grassy area outside high tunnel complex and from the perimeter of adjacent parking lot. Using XRF analysis, significant differences in concentrations of aluminum, manganese, phosphorous, potassium, rubidium, strontium, and titanium were found in the organic soils of the high tunnels, though concentrations did not exceed normal ranges or present concern for human health. These results are attributed to the use of overburden non-uniform urban soils in the establishment of the high tunnel complex. In future XRF evaluations for comparisons between elemental concentrations of soils, more sampling dates using in situ analysis would be necessary to construct baseline quantities that could then be monitored for changes within and between each system over time.

Ant communities of tropical forest canopies are relatively well described, but few studies have examined ant communities in temperate forest canopies. Likewise, the effects of urbanization on arboreal ant species richness and species composition remain unexplored. We examined the structure of arboreal ant communities at four sites along an urban-rural gradient near metropolitan Louisville, Kentucky. The University of Louisville (UofL) Belknap campus functioned as a representative urban green space, Iroquois Park as an urban park, Horner Wildlife Sanctuary as a suburban forest, and Bernheim Arboretum as a more continuous forest. In total we surveyed 439 trees with at least 99 trees sampled at each site. We examined how location, tree genus, tree size, and the presence of vines influenced the ant species richness and composition. Ant species richness was higher in Iroquois compared to Horner and UofL and increased marginally with increasing tree size. Ant species composition also differed across the locations, with the more rural sites tending to have greater diversity of species. Ant composition also differed among tree genera within a site. These results contribute to our knowledge of ant diversity in Kentucky, and further our understanding of the effects of urbanization on arboreal ant communities.

Cattle presence and beaver extirpation has led to incision, erosion, decreased vegetation, and eutrophication in many streams across North America, including Kimball Creek, a third order stream in western Colorado. These conditions have led to the localized extirpation of several native aquatic species, including the Colorado Cutthroat trout (Oncorhynchus clarki pleuriticus). For the last 20 years, beavers have been allowed to return and grazing pressure has decreased in the Kimball Creek system. Due to these passive restoration techniques, water retention and riparian vegetation have increased. I propose to investigate whether these habitat improvements have restored the stream to a condition capable of supporting a population of Cutthroat trout. Specifically, I will focus on how different levels of nutrients and shade affect water chemistry, primary production, and macroinvertebrate communities in beaver ponds. I will investigate various levels of nutrients and shade by using a mesocosm experiment that mimics natural conditions recorded in beaver ponds in Kimball Creek. I will then compare my results to the requirements of Cutthroat trout survival. Since the level of degradation observed in Kimball Creek is common in many small western streams, it is important to examine if small measures of passive restoration can allow the return of native aquatic species. I hypothesize that even in its degraded state, Cutthroat trout could survive in Kimball Creek.

Soils contaminated with nickel are a major human health hazard as many food crops, including legumes, and leafy and root vegetables readily uptake nickel. Consequently, consumption of food and water high in nickel can lead to major health issues, including cancer. Current soil remediation methods include physical (relocation/replacement), chemical (leaching, fixation, electro-kinetic removal, vitrification), and biological (phytoremediation, microbial and animal) of contaminated soil, which can be expensive, hazardous, and time consuming, sometimes taking decades to be effective. In order to address these issues, Sphagnum Peat Moss (SPM) was selected as a test sorbent for metal absorption due to its extremely low cost ($27-$69 per metric ton), sustainability, and high Cation Exchange Capacity (CEC). In this study, soil samples, collected from three Texas Superfund sites, were tested for heavy metal content and leachability to determine metal mobility in soil. SPM was added to contaminated soil (10% and 20%) to determine how much metal could be absorbed. The results showed a mixture of 20% SPM to soil immobilized 11% more nickel than the soil alone, showing the addition for SPM to nickel contaminated soils is an effective nickel absorbent over a short period of time (12 hours). This study showed that application of SPM to contaminated soils limits nickel mobility and has potential field application in the environment.

Plants defend themselves against attackers with a combination of chemical and physical defenses. A common feature of many plant species in lowland neotropical forests is a strikingly different color between new and mature leaves on the same plant, with young leaves often presenting with red or purple hues whereas mature leaves are dark green. This is true for both monocots (predominantly palm species) and dicots, leading to the hypothesis that young leaves are chemically but not physically defended. To test this, we collected new and mature leaves from 30 dicot species and 15 monocot species in a lowland tropical rainforest of the Peruvian Amazon. We measured leaf toughness (a measure of physical defense) with a leaf penetrometer three times on each leaf. Consistent with our hypothesis, young dicot leaves were significantly less tough than were their paired mature leaves. However, monocot leaves were remarkably tough independent of their age, with recorded toughness values exceeding 700 kPa (100 pounds per sq. inch) in some cases. These findings support the concept that young dicot leaves may invest heavily in chemical defenses to overcome limitations in physical defenses during leaf expansion as a means to deter herbivory, whereas monocot leaves invest immediately in physical defense. These divergent strategies may help shape plant communities in tropical forests.

Plant phenology is the seasonal development of the annual cycle of a plant, driven by temperature, photoperiod, and precipitation. Observations of plant phenology over time will clarify the interactions and feedbacks between climate changes and vegetation. The objective of this research is to examine the relationship between soil temperature and forest growing season length. The start and end dates of the vegetation growing seasons were determined using soil temperature data from two sites (Michigan and Kentucky). In addition, vegetation phenology was modeled using air temperature and the results were compared to determine if the soil temperature method based phenology is more accurate than the air temperature method. The effectiveness of those methods was assessed by comparing both soil and air temperature data results to observational data of Leaf Area Index (LAI) for the Kentucky site and phenology metrics derived from eddy covariance flux tower for the Michigan site. Satellite phenology data from the Advanced Very-High Resolution Radiometers (AVHRR) were also used and compared to the results. The data analysis so far predicts that the soil temperature method is more accurate than the air temperature method in predicting the vegetation growing season.

The experiment was conducted to demonstrate the behavior of fire propagation in wildlands using a matchsticks forest model. A model forest was designed on a flame resistant clay, on top of which matchsticks were inserted
and kept vertical to the ground by keeping space between them constant with the help of aluminum grid. The data for distance traveled by fire with time were taken at wide range of slopes from downhill of −25˚ to uphill of 45˚ on a
model forest of packing ratios 0.08 and 0.04. The minimum rate of fire spread was observed around 15˚ downhill. The data collected from this experiment follow tan^2θ and agree with the Rothermel's mathematical model of fire
propagation except at elevation above 35˚ for low packing ratio.

Population declines in amphibian species have been linked to a variety of factors, with the most significant being habitat loss and fragmentation. One potentially important contributor to habitat loss in Kentucky is strip mining. Soil and vegetation at reclaimed mining sites are typically altered versus sites that have not been mined, and soil from reclaimed mine sites is commonly acidic. The objective of this study is to look at the effects of soil acidity on microhabitat selection of salamanders post-metamorphosis. To do this, we observed the ability of spotted salamanders (Ambystoma maculatum) and Jefferson's salamanders (A. jeffersonianum) to detect, and select among a range of acidic environments (pH4-pH7) in experimental chambers. We hypothesized that salamanders will be able to select against acidic environments, suggesting that they may be able to avoid environments with high acidity during dispersal from their natal pools. Salamander behavior was recorded with time-lapse photography and data were analyzed using analysis of linear models. We found differences in the behaviors leading to microhabitat selection by each species, and the data suggest a general avoidance of acidic substrates. Among individuals within species, we observed high variability in the duration of behaviors contributing to selection of substrate microhabitat. In total, these data indicate that there is variation in the ability or willingness of individuals to discern differences in substrate acidity both at the intra- and inter-specific level. This suggests that conservation action aimed at reclaiming mine sites should manage soil acidity with the observed variation in mind.

Lichens are the result of a unique symbiotic relationship between a fungus and photosynthesizing partner. Due to their symbiotic nature, both the photobiont and mycobiont are able to provide its other species with essential needs. Lichens are commonly used as bioindicators of air quality as they obtain all their nutrients from the air due to the lack of roots. For this project, lichens were collected from a non-polluted site (Daniel Boone National Forest) and transplanted to four sites around Louisville. After five months at these sites, the samples were collected and sent to the University of Minnesota Research Analytical Laboratory to be analyzed for heavy metals. Our analysis of these results focused on eight heavy metals; Arsenic, Boron, Cadmium, Chromium, Copper, Mercury, Lead, and Zinc. We looked at each of these elements individually and combined per site and observed that Farnsley Middle School and Moore High School has the worst air quality, with 48.306 ppm and 47.314 ppm of combined heavy metals respectively. These sites are close to two major industrialized areas: Appliance Park and Rubber Town. In some cases, we observed that the control sample had a concentration higher than that of the transplanted sites. This led us to believe some elemental concentrations were observed to be lower due to the collection site having a higher amount of that element in the air than at the transplant site. The results of this study highlight the need to study lichen responses to air pollution throughout Kentucky.

Anthropogenic activities have lead to habitat degradation in streams throughout much of Western North America. These activities have resulted in a lower water table, loss of riparian vegetation, and higher water temperatures. The effects of habitat degradation on food quality and quantity for aquatic consumers could have large implications for food web stability as organisms move between different functional feeding roles. Kimball Creek, a degraded third order stream in Western Colorado, currently has one extant omnivorous native fish species, speckled dace (Rhynichtyus osculus) that primarily acts as an invertivore. Loss of riparian vegetation and increased nutrient loading by cattle has the potential to affect the functional feeding role of speckled dace by promoting a shift from invertivory to algivory. This shift in functional feeding role could lead to instability in the Kimball Creek food web and in turn affect restoration efforts. Through a mesocosm experiment that mimics the natural conditions of Kimball Creek, I propose to investigate how the current degraded conditions of Kimball Creek affect the functional feeding role of speckled dace in comparison to restorative conditions. I will focus on how nutrient additions and shade availability affect the quality and quantity of food sources for speckled dace, and the effects this has on the functional feeding role and growth rates of speckled dace in the Kimball Creek food web. By understanding the role omnivory plays in degraded aquatic systems, we can better understand the effect that efforts aimed at restoring these systems will have on their food webs.

Understanding larval fish phenology in dynamic systems provides managers with important indicators of sensitive fish life history patterns. Environmental factors are important for success of larval fish by influencing spawning and development. Water levels and temperature have been shown to influence the timing of the spawn in Kentucky Lake. Samples were taken within the southern 30 km of Kentucky Lake April-May of 2014, 2015, and 2016 to assess the larval fish community. Samples were collected using tandem larval pushnets (net=0.5m^2, mesh=1mm), fish were enumerated in the lab and identified to family. The timing of first detection occurred around the same period (April 17-April 25) in 2014, 2015, and 2016 for most taxa. Water temperature and headwater elevation were similar across these years and larval fish were detected within the same 9-day period each year. Using Kentucky Long Term Monitoring Program data from the last 30 years, we examined the patterns of water levels and temperature to detect changes in environmental variables. Changes in patterns of long term records could mean that the phenology of larval fish has changed and that resources available to these fish communities may differ over time. Phenology can be used as a harbinger of climate change and may be important for managing potential effects of invasive species.

Honey bees are the number one insect pollinator responsible for most of the biodiversity of flowering plant species. Honey bees are also crucial to agriculture as it produces honey, and pollenates a number of crops. For over ten years colony losses have been an issue in North America. Although there is no single cause of colony loss, many stressors are implicated including poor diet and exposure to chemical toxicity due to pesticides. In order to explore the effects diet and pesticides exposure have on honey bee health, we will provide three different diets of different quality (pollen, commercial bee food, and sugar), along with the mixture of two pesticides commonly used to protect crops from invasive insects. After 2weeks, from each treatment random bees will be selected to be used for RNA extraction. Quantitative PCR (qPCR) will be implemented to measure the expression of genes known to be involved in detoxification and oxidative stress caused by chemical poisoning, and nutrition responsive genes.

Sunlight provides the essential energy required for all growth, reproduction, and defense in most plants. In neotropical forests, canopy trees have leaves in close proximity to one another that are exposed to full sunlight or almost completely shaded. The purpose of this study was to assess whether damage levels to leaves differed between sun and shade leaves of neotropical canopy trees. We measured physical damage to leaves in direct sunlight compared to shaded areas of five canopy tree species (individual trees) in the Peruvian Amazon, from an existing canopy walkway. For each tree, we estimated damage on approximately 250 leaves in each of direct sunlight and shade using a previously published visual assessment technique. Overall, we found average damage levels of 11.4 +/- 6.3% and 18.0 +/- 18.8% on sun and shade leaves, respectively. One of the five tree species having 50% damage in shade leaves, had a contrast of over 1000% difference between sun and shade leaves, even after subsequent consideration of leaf age. Our results suggest that canopy trees in a neotropical lowland forest have variable effects of sunlight impacting their susceptibility to herbivory, which implies that natural history of each tree species is an important determinant shaping species-specific plant-herbivore interactions.

Expansive road systems have a negative effect on wildlife populations through increased mortality by animal-vehicle collisions. In the United States, an estimated one million vertebrates are killed each day on public roadways. It is unclear what percentage of the mortality, especially for small animals, is due to intentional collisions. We tested three hypotheses: 1) drivers who intentionally hit wildlife possess species bias, 2) drivers of trucks are more likely to intentionally hit wildlife than drivers of cars and SUVs/vans, and 3) intentional wildlife-vehicle collisions are positively correlated with the distance from Murray city center. A realistic model of a snake and a turtle, and a red cup were used for our study. We selected 45 mph roadways located varying distances from the city center. Models and roadways were randomly selected and data on 50 vehicles were collected per trial. We compared model type, vehicle type, and distance from city center to the number of intentional vehicle hits to determine the presence of driver bias. Snakes were hit by vehicles significantly more often than turtles or cups. The variance between vehicles hitting turtles and cups was not significant. Vehicle type had no relationship with number of intentional hits. Information gained from our study spotlights biases in intentional wildlife-vehicle collisions. Public educational efforts and wildlife crossings structures need to emphasize snakes because the snake model was targeted significantly more often than the turtle model.

Phenology is nature's calendar; it describes the seasonal timing of organisms that synchronize their lives to cyclical climatic changes throughout the year and across years. Phenology varies with latitude and other drivers of temperature and climate. Hardiness zones were established by the USDA as a basis for comparing phenology across North America. Hardiness zones are defined by the average annual minimum winter temperatures for any given location. We set out to evaluate similarities and differences in the first bloom date of common lilac (Syringa vulgaris L.) and red maple (Acer rubrum L.) across various locations within the same plant hardiness zones. We used the US National Phenology Network (US-NPN), a citizen-scientist observational database, which dates back to 1951 and the Pan European Phenology Database (PEP) which dates back to 1868. With the help of ArcMap, we were able to separate each data point according to its location within its respective hardiness zone. Results showed there is a significant difference between the zones in the long-term US-NPN data for common lilac. Evaluating the first bloom dates of common lilac and red maple will help us understand if hardiness zones are an accurate representation of phenological changes and the degree to which they can help us understand future plant phenology work.

Urban karst lakes are understudied with respect to the role they play in urban karst hydrology and the water quality implications from their existence in the landscape. It is well documented that stormwater runoff influences various parameters (heavy metals, oil and grease, salts, etc.) and that other urban impacts can negatively impact groundwater systems. Additionally, these lakes can serve as accumulators of sediment and waste that is directed into the system. Here, we present data from Limestone Lake, a karst lake in the urbanized area of Bowling Green, Kentucky, regarding the extent of impacts to its water quality and role in the heavily modified urban karst system. Ten-minute resolution data were collected for water depth, pH, conductivity, turbidity, and temperature over a four-month time span. Weekly water samples were collected for oil and grease, biological oxygen demand, alkalinity, chemical oxygen demand, anions, cations, total organic carbon, total fecal coliform, and E. coli. The results indicate the lake is responsive to stormwater pollution with a lag time. Changes in the water geochemistry values, water level, and parameters, such as nitrates and E.coli, resulted from storm events. High-resolution monitoring displayed daily and seasonal fluctuations. Seasonal and daily data indicate that the lake is not as contaminated as nearby groundwater sample sites and may function more as a surface water body in the karst system. The importance of karst lakes and their role in urban karst hydrology merits further research and this case study proves that additional site-specific knowledge is still needed.

Is aboveground pathogen load influenced by belowground pathogen presence, plant community diversity, or precipitation amount?

Interactions between plants and their aboveground and belowground pathogens are often highly intertwined and difficult to tease apart. Belowground microbes, like arbuscular mycorrhizal fungi and ectomycorrhizal fungi, can form mutualistic relationships where both the fungi and plants benefit. However, a plant may have natural enemies both above- and belowground. We tested whether plant diversity (species richness, phylogenetic diversity), the presence or absence of soil biota, and water availability interactively affect plant susceptibility to damage by aboveground enemies such as foliar pathogens and herbivory. We examined experimental tallgrass prairie communities at Tyson Research Center composed of 17 Missouri native species. These communities experienced drought or well-watered conditions and were planted in sterile soil or soil inoculated with microbiota from a tallgrass prairie at Shaw Nature Reserve in 2014. The communities consist of either one or six species, and the six-species mixes are either closely or distantly phylogenetically related. During the growing season, we recorded the type and severity of damage present on aboveground parts of plants. Our study shows that soil microbes influence the degree of aboveground damage that plants experience, however results varied by species. We conclude that above- and belowground plant-pathogen interactions are diversified between species, and that drought affects some species more drastically than others. Due to global warming, some ecosystems, including tallgrass prairies, may experience more frequent or intense droughts in the future. By discovering the conditions under which certain species experience the lowest pathogen load, we can better replicate favorable conditions in restoration and conservation projects.

Insect pollinator populations, which are crucial for maintaining ecosystems, have been steadily declining across the globe in recent decades, a trend that is intensified in cities. Urban native plant gardens could potentially offset these losses. However, the impact of these urban gardens on the conservation of native pollinators and other insects is not yet fully understood. This research attempts to determine the composition of insect communities in native plant gardens using an urban gradient approach. Across the city of Louisville, KY, seventeen garden sites and two reference meadows were selected. In each of these, bowls of three colors (blue, white, yellow) set at two heights (15 cm and 100 cm) were used to capture insects for identification to taxonomic order. The impact of bowl color, bowl height, garden characteristics, and degree of “urbanness” (measured by % Impervious Surface) on abundance and diversity of insects captured were assessed. We found that yellow bowls captured the most insects, while height played a slightly less significant role, with the high bowls capturing more. At the order level of taxonomic resolution, we found no statistically significant effect of garden characteristics or surrounding impervious surfaces on insect abundance or diversity. However, native plant gardens captured as many or more insects as the meadow reference sites, suggesting that the creation of more native plant gardens may well be a viable conservation strategy for supporting insect species, including pollinators, in urban environments.

“SENSE” is a new research program funded by NSF-EPSCoR (2016-2020) that builds on previous cyberinfrastructure to address water quality issues shared between Kentucky and West Virginia. Our focus is on the toxic cyanobacteria (cyano-HABs) that impact drinking water, human health, wildlife and livestock, aquatic life, irrigation sources, and energy production.

Amphibians worldwide are experiencing declines, including the crawfish frog (Lithobates areolatus), an IUCN red list Near Threatened species and Species of Greatest Conservation Need in the state of Kentucky. The crawfish frog is primarily fossorial and relies solely on crawfish burrows for summer retreats and winter hibernation sites. Because of its secretive nature, calling surveys of male vocalization during the breeding season are the ideal method to monitor crawfish frog populations. However, the peak-breeding season often only lasts a few nights, sometimes even a single night, making the timing of these surveys critical to accurately estimate populations of these frogs. We deployed frogloggers at 7 wetlands from February 27th to April 23rd, 2016 in the West Kentucky Wildlife Management Area, McCracken Co. Kentucky. Loggers recorded the first 5 minutes of every hour of every day during the sampling period. The first minute of every recording was analyzed for species calling and abundance rank of each species using the North American Amphibian Monitoring Program's Protocol. Preliminary results show that crawfish frogs called from February 27th to March 17th. Mean calling frequency was highest between the hours of 12am and 6am and 6pm to 11pm. These preliminary results suggest that the best time to conduct calling surveys for crawfish frogs is from late February to mid-March between the hours of 12am and 6am and 6pm and 11pm.

Trophic cascades are important in structuring diverse ecosystems, from terrestrial to aquatic systems. However, trophic cascades vary in strength both within and among ecosystem types. Recent theoretical and modeling research suggests that intraspecific variation could be a factor in varying strengths of trophic cascades. In particular, body size variation within a species has implications for species' prey choice, behavior, and niche breadth.

In this project, we manipulated the body sizes in populations of mole salamanders (Ambystoma talpoideum) to investigate the role of intraspecific variation in a top predator on trophic cascades. A. talpoideum are aggressive predators in fishless ponds and exhibit a wide range of body sizes due to their unique paedomorphic life history. We hypothesized that populations of similarly sized A. talpoideum would exhibit strongest trophic cascades due to overlap in niches and predation on similar prey items. As body size variation in a population increased, we predicted reduced top-down predation pressure due to smaller overlaps in niches and cannibalism. To test these questions, we constructed 54 1000-L mesocosms at Hancock Biological Station in Murray, Kentucky, which were populated with one of three salamander size-structure treatments. We monitored changes in the ecosystems through a suite of abiotic and biotic parameters over a period of six months.

Results show that presence of salamanders significantly increased chlorophyll-a concentrations, suggestive of a trophic cascade. Preliminary results suggest that the cascade is not mediated by a traditional pathway through zooplankton and macroinvertebrates, but instead by tadpoles which act as strong herbivores and limit concentrations of primary producers. These tadpoles were opportunistically deposited to the mesocosms by ovipositing female tree frogs, which have been shown to selectively oviposit in ponds that do not contain A. talpoideum. This suggests that A. talpoideum could mediate a trophic cascade via indirect fear-based interactions.

Large flood storage reservoirs are often the target of habitat enhancement projects because they lack aquatic structure due to dramatic shifts in hydrology. State agencies and anglers often add artificial structures to these reservoirs to attract fish and providecover for reproduction, spawning, and refuge from predation. However, some structures may be short-lived, costly, and unaesthetically pleasing in shallow water. Bald cypress (Taxodium distichum) seedlings have been successfully grown in areas that suffer from habitat loss in Kentucky Lake.The growth of these trees is tolerant of flooding. This study compared fish use of bald cypress and stake beds, which are a common type of artificial structure in the lake. Boat electroshocking was used to compare the fish assemblages among structure types. Open water areas were used as a control. Sites were sampled five times from April-July, 2016. Samples included 186 fish representing 24 species. Sunfish (Centrarchidae) comprised approximately 50% and 54% of fish collected at stake beds and bald cypress trees, respectively. Clupeids and Cyprinids comprised approximately 70% of fish caught in open water. Repeated measures ANOVA revealed no significant differences in catch-per-unit-effort (CPUE) or species richness over time among stake beds, bald cypress, and open water. A comparison of overall CPUE and species richness revealed a higher species richness at stake beds, but no difference in CPUE between the structure types. Results show that bald cypress can concentrate fish as effectively as stake beds in shallow areas of embayments, providing a cost-effective and sustainable way of enhancing aquatic habitat.

Plants are able to chemically respond to volatile cues from nearby plants experiencing a stress such as herbivory or pathogen invasion. For example, the specific odors of freshly mown grass, called “Green Leaf Volatiles” (GLV), are known to trigger defense responses in undamaged plants. However, little is know about how the dose of the volatile affects the severity of the response. We hypothesized that the degree of defense would be directly proportional to the concentration of the volatile cue. To test this, we used the GLV cis-3-hexenyl acetate (z3HAC) and the model plant Arabidopsis thaliana. We grew Arabidopsis in a growth chamber and performed a factorial experiment with seven different concentrations of z3HAC, where half of each concentration group was given oral secretions of Spodoptera exigua, the beet army worm. The oral secretions were intended to generate an induced resistance response. Then, we measured the relative growth rate of a naive cohort of S. exigua caterpillars. Volatile dose had an inconsistent effect on caterpillar growth rates that did not reflect our predicted dose-dependent resistance profiles. However, oral secretions did generate resistance in the form of reduced growth rates overall. Our results suggest that dose of a cue is important, but the relationship between dose and response deserves further study.

Environmental DNA (eDNA) provides an effective, non-invasive method to determine organism presence or absence in an efficient manner. The majority of salamanders native to central Kentucky have an aquatic phase to their life cycle. Many Ambystomid species persist as aquatic larvae for just a few months before progressing to their terrestrial phase. We developed species specific eDNA primers for streamside (Ambystoma barbouri) salamanders that effectively amplify salamander DNA filtered from stream water. We collected 1 liter water samples biweekly from February to July 2015 in four small streams in Jessamine and Madison County, Kentucky to examine season fluctuation in eDNA levels of different salamander species. Initial data reveal a complete absence of A. barbouri eDNA in early spring samples but high levels later in the spring corresponding with breeding and larval presence. RT PCR analysis is in progress in order to determine quantitative levels of DNA at each collection. These data add to the growing pool of knowledge concerning eDNA monitoring of species and should provide useful reference data for future monitoring or range delineation studies.

Urban spaces have been considered wildlife deserts, but recent studies have suggested that urban areas can serve as wildlife habitat if suitable spaces are designed and included in cities. Urban gardens may be particularly valuable for pollinators, which are often capable of flying long distances among food patches and can have relatively small space requirements. We investigated whether urban native plant gardens and parks can help support populations of native butterflies. Specifically we used clay models of large and small caterpillar to assess caterpillar survival rates in 20 sites in Jefferson County, Kentucky. Our sites included park meadows and public and private gardens that varied in age, area, and the amount of impervious surface surrounding them, which is a measure of 'urbanness.' We identified predator taxa (arthropods, birds, mammals) by marks they left in the clay caterpillars. After one week, on average 78.1% of large caterpillars and 63.1% of small caterpillars had been attacked. Large caterpillars were significantly more likely to be 'predated' primarily because they experienced higher predation by arthropods. Interestingly, garden area, garden age, and impervious surface around sites did not affect caterpillar survival, and the predation rates we observed were similar to those from studies in natural areas. These results suggest that even small, relatively new urban gardens have the potential to serve as effective pollinator habitat. As plantable residential lands often comprise a large proportion of urbanized spaces (30% in Jefferson County), encouragement of native plant gardens could have a large positive impact on pollinator populations.

Zebra mussels (Dreissena polymorpha) are an invasive bivalve that have become established across much of the United States. These filter feeders compete with native aquatic organisms for food and space by directly consuming detritus and phytoplankton and by attaching directly to solid surfaces via byssal threads. Zebra mussels were reported in the Ohio, Cumberland, and Tennessee Rivers in the early 1990's, but densities in Kentucky Lake were low and few observations of juveniles suggested limited reproduction. However, in early 2017, several observations of high densities of zebra mussels (including juveniles) were reported in Kentucky Lake. We sampled zebra mussels along transects that included shoreline (n=39) and offshore samples (n=18) from the lower 40 km of Kentucky Lake during August – October, 2017. Densities ranged as high as 7284/m2 and averaged 1022 ± 907/m2 and 1210 ± 1252/m2 for shoreline and offshore samples, respectively. Zebra mussels generally require dissolved inorganic calcium levels greater than 20ppm. Past research has shown dissolved calcium concentrations in Kentucky Lake in the range of 12-15ppm and too low for zebra mussel establishment. However analysis of long term data show increases in alkalinity (calcium carbonate), chloride ions, and pH which indirectly suggest that conditions in Kentucky Lake may have reached tolerable thresholds for zebra mussels. Anthropogenic additions of road de-icing brine (calcium chloride) may be increasing elemental concentrations in Kentucky Lake. Establishment of significant zebra mussel populations in Kentucky Lake could have drastic ecological and economic effects.

Environmental DNA (eDNA) provides an effective, non-invasive method to detect the presence of rare organisms in aquatic systems, provided sufficient molecular tools are available. Percopsis omiscomaycus is a small fish with a limited, disjunct distribution in central and eastern Kentucky. We amplified and sequenced a 769 BP region of Percopsis omiscomaycus cytochrome b and used this sequence to design eDNA primers that selectively amplify P. omiscomaycus DNA from filtered water samples. One liter water samples were collected from 17 locations in northeastern Kentucky, filtered, and DNA was extracted in a manner consistent with established methods. Additionally, each location was intensively sampled for P. omiscomaycus by seining. eDNA levels quantified using a primer/probe assay and real time PCR only partially correspond to field collection data, perhaps providing additional insight into P. omiscomaycus distribution. These data add to the body of knowledge concerning P. omiscomaycus distribution and provide a useful tool for detecting cryptic populations for this and other species.

Keegan Thompson, Alyssa Y. Stark, Stephen P. Yanoviak

Department of Biology, University of Louisville, Louisville, KY 40292

Tree-dwelling (arboreal) ants exhibit a remarkable ability to cling to a broad range of substrates in their environment. Presumably, the asperity (roughness) of substrates like leaves and tree bark affects the adhesive performance of ant tarsi. We explored this relationship by measuring the adhesive performance of ants on oak leaves differing in trichome density. We measured normal (orthogonal to the surface) and shear (across the surface) adhesion loads for 72 Aphaenogaster rudis workers on leaves of Quercus bicolor (875 trichome per cm2) and Q. rubra (9 trichomes per cm2). Normal adhesion was zero for all ants on both species of leaves, whereas shear adhesion loads ranged from 0.05-0.20g on Q. bicolor and 0.00-0.10g on Q. rubra. There was a tendency for adhesive load to increase with ant mass, but this pattern did not confound the main effect of trichome density. These results suggest that ants can cling to hirsute leaves even when subjected to large lateral forces (i.e., a leaf fluttering in the breeze) but are likely to fall from leaves (regardless of trichome density) when exposed to vertical forces. Ultimately, such patterns potentially explain the effects of natural substrates on the foraging patterns and local distributions of common arboreal ants.

Leafcutter ants are conspicuous components of neotropical forests. Workers of various species cut leaves and bring fragments back to their colony. Small worker ants (hitchhikers) will periodically appear on the leaf fragments carried by larger workers (carriers), however many aspects of this behavior remain unknown. The objective of this study was to determine if hitchhiker size correlates to carrier size. I measured hitchhiker and carrier size of 30 leafcutter ants found along a trail in the Peruvian Amazon. Although there was no difference in size of carriers with and without hitchhikers, hitchhiker size showed a tendency to increase with carrier size (R2=0.15). An appropriate next step would be to explore the relationship between carrier and leaf size.

Since 2006, over six million bats have been killed by White-nose Syndrome (WNS). With nine species in rapid decline, there is reason to believe that niche partitioning will make it more difficult for these species to reemerge because of competition for food. Our goal was to find if the change in bat communities post-WNS influenced the diet of evening (Nycticeius humeralis (Rafinesque)) and tri-colored (Perimyotis subflavus (F. Cuvier)) bats, and to what extent. The evening bat is not susceptible to WNS and the tri-colored bat is WNS-susceptible. We obtained pre-WNS data from a bat study completed in 1996 at Land Between the Lakes (LBL) and compared it with current data on bat diets. Sticky traps were used to determine insect availability, while guano was dissected to determine the insect prey eaten by each bat species. The guano was spread across a petri dish and insect order occurrence recorded by percent volume. Before WNS, evening bats consumed 31% Coleoptera, 14% Diptera, 23% Lepidoptera, and 17% Homoptera. Tri-colored bats ate 19% Coleoptera, 26% Diptera, and 52% Lepidoptera before WNS. After WNS, the occurrence of Coleoptera in the guano of both bat species was more than double pre-WNS levels while the occurrence of Lepidoptera was markedly lower. The change in diet of evening and tri-colored bats may be explained by competitive release following high mortality of WNS-susceptible species. Increasing overlap in diet at the ordinal level between the evening and tri-colored bat may indicate increased competition between evening and tri-colored bats.

The Clean Water Act (CWA) is United States (U.S.) federal law protecting U.S. surface waters from pollution. The Environmental Protection Agency (EPA) is the enforcer of CWA tenets and shows latitude in its jurisdiction helping states if necessary to meet CWA goals. Yet, the CWA authorizes states to design and execute programs to meet its goals. Benthic macroinvertebrates are sensitive to water pollution; they are real time living indicators of water quality. Two stream reaches of the Lee Branch on the Midway University Campus are surrounded by natural terrestrial habitat. In accordance with the CWA, we wanted to determine the health of the Lee Branch on our campus by performing biological stream assessments and stream habitat assessments of these reaches during the summer, 2016. We identified aquatic habitats as riffles, runs, pools, and glides and generally randomly sampled each. Microhabitats within habitats were randomly sampled when appropriate. We sampled for benthic macroinvertebrates by kicking substrates into a D-frame net, with contents emptied into buckets and typically sieved in the lab. Kept specimens were preserved and identified; crayfish were released. We formulated integrity ratings from biotic indices and habitat ratings from habitat scores allowing categorization of stream reach conditions. Reach 1 rated fair for both integrity and habitat; when contiguous upstream and downstream habitats were included, habitat rating increased to good. Reach 2 rated fair for both integrity and habitat. Water chemistry at benthic macroinvertebrate collection sites with two exceptions showed both reaches within Kentucky's acceptable ranges for aquatic life.

Estimating fish abundance holds great importance for freshwater ecology and fisheries management, but current techniques can be expensive, time consuming, and potentially harmful. Environmental DNA (eDNA) has proven an effective and efficient technique for presence/absence detection of freshwater vertebrates. Additionally, recent studies report correlations between total fish biomass and eDNA levels although few studies have examined this relationship in small, lotic systems. The present study examines the relationship between biomass and eDNA concentrations of smallmouth bass (Micropterus dolomieu) and greenside darters (Etheostoma blennioides) in central Kentucky streams. Population estimates were completed in the lower Kentucky River watershed in streams of varying size through the utilization of electrofishing and three pass depletions. eDNA samples were collected concomitantly, and DNA levels were quantified using primer/probe assays and real-time PCR. This data should be useful in determining the effectiveness of eDNA in lotic systems.

Effects of Roundup on Feeding Behavior of Larval Blue Dashers, Pachydiplax longipennis. KAYLEEN K. PARKER and CLAIRE FULLER, Biology Department, Murray State University, Murray, KY 42071.

Herbicides are widely used and prior research has shown that many herbicides are harmful to organisms and the environment. Herbicides enter aquatic environments through runoff or aerial dispersion from fields. The objective of this study was to determine if Roundup (active ingredient Glyphosate) causes negative effects on feeding behavior of larval Pachydiplax longipennis. Larvae were captured from rainwater-filled mesocosms at Hancock Biological Station in Murray, KY. Larvae were exposed to one of four concentrations of Roundup (0mg/L, 2.5mg/L, 5mg/L, or 10mg/L). Daphnia consumption trials were conducted at 7 and 14 days post-exposure. There were no significant differences among treatments for whether or not larvae ate offered Daphnia for Day 7 (2 =0.597, df =3) or Day 14 (2 =0.178, df =3). For the trials on Day 7, Roundup concentration did not have a significant effect on the time it took the larvae to consume 1, 2, or 4 Daphnia; however, exposure significantly increased the time it took the larvae to consume three Daphnia (P=0.036). For the trials on Day 14, concentration did not have a significant effect on the time it took to consume 1 or 2 Daphnia; however, Roundup significantly increased the time it took to consume 3 (P=0.010) or 4 Daphnia (P=0.029). Thus, Roundup slowed prey consumption, suggesting that it could have a negative impact on larval dragonfly predation and, consequently, their overall quality of life. More time to capture and consume food suggests higher energy usage during predation.

Dams have long been know to influence biota, including freshwater mussels, and their ecology downstream primarily due to the significant alteration of flow and temperature regimes. Key ecological attributes that may be affected include growth, reproduction and recruitment and nutrition. The Green River is one of the most biologically significant river systems in the United States. Historically the Green River boasted over 70 mussel species, including several federally listed as endangered. Mussel diversity and abundance is known to be significantly higher with distance from the Green River dam. Mussels have been regularly used as an indicator of stream health and it has also been assumed that individual mussel health and condition improves downstream. Water samples were collected and analyzed for concentrations of certain elements to show a possible correlation between mussel condition and distance from the dam. Our investigation examined whether health and condition of two common mussel species, the mucket (Actinonaias ligamentina) and three ridge (Amblema plicata), varied between upstream and downstream populations based on a mussel condition index and relative erosion of the shell periostracum.

Symbiotic interspecies relationships are common in the tropics, where high levels of biodiversity have enabled many opportunities for species to coevolve. In some instances, however, the ecological parameters of these relationships remain unclear. For example, some plants in the family Melastomataceae house ants that emerge from hollow domatia at the petioles when the plant is disturbed. Assuming that these domatia-inhabiting ants help protect the melastome in exchange for shelter, we predicted that melastomes with domatia suffer less leaf damage than melastomes without domatia. To test this, we located over one-thousand leaves from sixty melastomes at the ACTS research station near Iquitos, Peru, and assigned each leaf to one of seven categories based on the percent leaf damage. We then compared damage percentages between melastome leaves with and without domatia. In contrast to our hypothesis, our analysis discovered no significant damage difference between the two types of leaf, suggesting that the presence of domatia has no effect on melastome herbivory. These results expose the need for further research to determine whether domatia-inhabiting ants provide any service to their melastome hosts, and if not, why some melastome species invest in domatia while others do not.

Amur honeysuckle (Lonicera maackii) is the most important invasive species in the Ohio River Valley. Previous work has shown extensive dieback of honeysuckle in the region, coupled with the appearance of the native fungal pathogen, honeysuckle leaf blight (Insolibasidium deformans). Our goal was to find if the blight causes growth decline or mortality. Seedlings were grown under greenhouse conditions. Blighted leaves were collected from the field, and treated seedlings were sprayed with a spore solution. Treated and control plants were placed into a growth chamber with conditions set for optimum spore growth (~100% relative humidity and 16°C) and then returned to the greenhouse after leaf blight began to develop. Growth and dark-adapted chlorophyll fluorescence were measured periodically, and blighted leaves had greatly reduced leaf area, chlorophyll fluorescence, and relative growth rates. A repeated-measures analysis of aboveground growth indicated that larger, more quickly growing plants were more likely to be infected, but their growth rates were subsequently reduced much more than uninfected plants. No infected plants died, but this experiment supports our hypothesis that leaf blight causes a significant growth decline in Amur honeysuckle.

Green roofs, or living roofs, are a sustainable building practice that are implemented mainly in cities across the world to reduce the urban heat islands, manage storm water run-off, and increase the efficiency of building by absorbing heat. These roofs provide synergistic benefits to energy and environmental building design as a single entity. In addition, green roofs increase the efficiency of the systems inside the building, improve air quality, and better stabilize ecosystems of native and adaptive species by providing an elevated habitat. Green roofs are able to replace the vegetated land that was removed to construct the building. Consisting of insulation, a waterproof membrane, a drainage layer, a soil mix, and a choice of native plant species, green roofs allow a building to stay insulated and absorb water while still maintaining the well-being of the building.
Since green roofs are seen as a sustainable building practice, they are recognized in LEED®. LEED stands for Leadership in Energy and Environmental Design and was developed by the United States Green Building Council. As a result of green roofs providing synergistic benefits, they capture many varying degrees within the LEED rating system while allowing a building to be resource-efficient throughout its life. One can maximize their LEED certification using green roofs by reaching LEED categories including Sustainable Sites, Water Efficiency, Energy and Atmosphere, and Materials and Resources.

Abstract
At the University of Kentucky Center for Applied Energy Research, recent efforts have been made to optimize and reduce the cost of a unit designed to control an automated photobioreactor system in which microalgae is grown to reduce CO2 emissions. In contrast with a previously used system based on a compactRIO controller and NI LabVIEW software, a new optimized version utilizes an inexpensive Beagle Bone panel as a microcontroller that can be programmed through the use of Python software. Notably, the cost of the new control system was 1/10th of the price of the previous unit.

The purpose of this project was the research, design, and fabrication of a biomass production and feeding system for the application of alternative heating methods for small structures on campus. The biomass material to be utilized consisted of the waste sawdust produced by college industry. The research group reviewed existing literature and current manufacturing practices. Samples of production grade materials were obtained for comparison testing with regard to energy potential. The current project concluded with the design and manufacture of a functioning prototype for initial pellet production for testing. Future research will continue to revise the prototype while producing viable comparison material for further data collection and analysis prior to implementation.

Carbon nanotubes (CNTs) have received great attention due to their exceptional multi-functional properties (thermal, electrical, and mechanical properties). However, CNTs exhibit superior properties only along the axial direction due to the presence of strong covalent bonding in the carbon plane; they exhibit poor properties in the transverse direction since there are only weak van der Waals interactions among individual nanotubes. Therefore, the CNTs are essentially one-dimensional (1D) nanomaterials and their applications have been somewhat limited. A novel carbon nanomaterial has been proposed recently: the three-dimensional carbon nanotube/graphene hybrid (3D CNT/Graphene). The 3D CNT/Graphene is consisted of vertically aligned carbon nanotube pillars interconnected by horizontal graphene layers. This unique hybrid architecture would possess desirable transport and mechanical properties in the transverse directions (along the graphene planes) while maintaining the excellent properties in the axial directions (along the nanotubes). Unlike conventional materials whose microstructures are relatively “fixed”, these new 3D nanomaterials are highly “tunable” from the structure standpoint. Through this project, an Integrated Computational Material Engineering (ICME) approach was used to design and optimize the 3D CNT/Graphene hybrids. Various representative models of the 3D CNT/Graphene have been constructed and analyzed by using the finite element method. Results show that the axial modulus of the 3D CNT/Graphene increases exponentially with respect to the graphene thickness while the transverse modulus remains constant. The optimal design of 3D CNT/Graphene requires a minimum normalized thickness of g/t>5, where g and t are the thickness of horizontal graphene and vertical nanotube, respectively.

This project focused on the development of MATLAB code which was designed to process images of aluminum castings for micro-pores that occurred during solidification. Initially, the code was created to identify the pores in the images by selecting areas that were darker than their surroundings; pore identification was signified in MATLAB with a ring around the fault. This program was then refined so that the code was able to identify faults while ignoring dark bands at the edges of the images. Additional refinements allowed for the grouping of faults that were located within a certain distance of each other. Clusters of pores were then identified with rings of the same color around their constituent pores so that each cluster could be differentiated from the others. Finally, the code was further refined to prevent large, clearly-individual pores from being added to clusters, and to limit the chaining of pores to a specified radius from the central-most pore so they couldn't aggregate into one large cluster across the whole image. Ultimately, the code was able to characterize pore clusters in terms of their sizes, numbers of pores and average distances, in order to identify those clusters at which fatigue cracks were nucleated in the cast aluminum alloys, as a critical step towards accurate life prediction of the alloy in engineering applications. While micro-pore clusters have been recognized as the preferred fatigue crack nucleation sites, the quantitative relationship between these pore clusters and crack nucleation had not been established previously.

In this study, Landsat-8 imagery of west Lancaster, Pennsylvania is examined using different remote sensing techniques to detect agriculture conservation practices. Farmers in this study area practice conservation practices known as contour farming and strip-cropping, where they grow alternate crop strips down a slope on the contour to reduce soil erosion and runoff. It is valuable to map and monitoring the cropland where the conservation techniques are applied. Remote sensing provides the advantage of covering a large area with regular revisit frequency. Two different methods are experimented in this study for their effectiveness of detecting conservation practice using Landsat-8 OLI image. The Image was collected in August 8, 2015. Normalized Difference Vegetation Index (NDVI) is derived from the original dataset to enhance the difference in crop strips. Results based on an unsupervised classification of the NDVI image and high pass filters are compared for detecting the use of contour farming methods.

Unmarked Burial Identification at the Frankfort, KY Cemetery using Ground Penetrating Radar and LIDAR data through ArcGIS
Jarod Jones, Jordan Wilson, Jeremy Sandifer, Ken Bates
In many cemeteries of the Commonwealth, older sections delineated for those of the socio-economical challenged may contain unmarked graves. This occurs for a number of reasons, including lack of accurate historical records, changes in cemetery boundaries over time, and headstone displacement. Many of these places are of significant historical and cultural value, especially from a genealogical perspective. The most common methods of probing and exploratory excavation are invasive, labor intensive, and time consuming and highlights the need for alternative methods for identifying unmarked graves. In this current project, the use of ground-penetrating radar (GPR) and light detection and ranging (LiDAR) is used successfully for identifying unmarked graves with a high rate of consistency. GPR and LIDAR results are overlaid using geographic information system (GIS) in order to assess the spatial correlations in signal variations in each dataset. The GPR unit used did not contain a spatial component, however a handheld GPS device was used in order to verify the exact placement. Preliminary results indicate that while the GPR is very effective at identifying potential unmarked burial locations, it is expensive and requires an onsite expert to definitively interpret the results. The effectiveness of the LiDAR derived elevation measurements are less conclusive.

Kentucky, a major agriculture state, has rapid population growth during 1990 to 2010 by 17.74%. Five major watersheds of Central Kentucky viz. Pond Creek, Bear grass, North Elkhorn, South Elkhorn, and Hickman Creek were selected for land cover change assessment. The United States Geological Survey Classified standard 30 m resolution National Land Cover Data (NLCD) of 1992 & 2011, and hydrological unit for watershed boundaries were analyzed to find out the percentage change in the land cover classes in each watershed. The developed area was significantly increased with maximum percent increase in Bear grass watershed i.e. 16.44% and minimum in North Elkhorn by 12.21%. However, the forest and cultivated areas were significantly decreased in all watersheds with maximum percent decrease for forest by 9.9% in Pond Creek and cultivated area by 11.38% in Hickman watershed. With this result, we can conclude that urbanization is increasing rapidly resulting a loss of forest and cultivated land. The rapid increase in urbanization with increase in population, on cost of forest and agriculture area, will make us vulnerable towards hazards and others. This study suggests for further research on impacts of these land cover changes on water quality, urban land management, and ultimately on quality of life.

Landslides are a well-known but costly natural hazard in Cincinnati & Northern Kentucky, but are difficult to catalog and monitor because of the steep forested slopes that characterize the region. Using elevation change maps derived from successive LiDAR surveys from 2007 and 2012, locations of previously reported and cataloged landslides were observed, and new uncatalogued landslides were searched for. In the initial study area representing 1.6% of Kenton & Campbell counties, six out of ten previously cataloged landslides showed signs of activity, and eight previously uncatalogued landslides were identified. Both thin colluvial slides and slumps were identified using this method, and previously uncataloged landslides were either field checked or confirmed using air photos in Google Earth. The use of sequential LiDAR in this area of heavy vegetation and steep slopes appears to be a useful tool for monitoring known landslides and for delineating and cataloging new landslides, and will support further study into the character of slope movement in the region.

The storage of soil organic carbon (SOC) at depths greater than one meter in valley bottoms, and processes operating on that SOC, are not well understood. We examine the stock of SOC within alluvial landforms: floodplains, terraces, and bars, in the Clarks River National Wildlife Refuge. Preliminary data from cores along a transect from a terrace to an adjacent channel bar were collected to depths of 1 to 1.5 meters. Bulk density, particle size, and loss-on-ignition were used to estimate SOC in each landform type. Average subsoil-SOC stocks for each landform decreased towards the river channel, with the largest value of 1.8 kg/m^2 ± 0.3 SD found in the terrace and the lowest value of 0.9 kg/m^2 ± 0.18 SD found in the near-channel bar. The floodplain had an average value of 1.104 kg/m^2 ± 0.4 SD. This variability is likely due to differences in soil texture, where terraces have greater soil development due to infrequent flooding which allows for clay accumulation, a predictor for SOC storage at depth. Although floodplains and near channel features observe greater flooding and potentially greater deposition and sequestration of SOC, this may be stored temporarily and oxidized with increasing residence time. A buried soil observed within one profile had a higher total C stock than the overlying surface soil. Additional transects and measurements of SOC via combustion and gas chromatography will be used to test these observations.

Sediment stored in floodplains and low alluvial terraces along the middle Tennessee River reflects flood frequency and magnitude during the past ca. 1500 years. This study uses the stratigraphy, sedimentology and geochronology of three alluvial terraces to infer paleoflood deposits. Buried soils at the three locations are older than 630 CE and suggest a multi-century period of landscape stability. Multiple flood deposits are separated by weakly developed soils, indicating an increased flood frequency until ca. 1910 CE. Optically-stimulated luminescence dates of flood deposits yield ages of 580+/-110, 835+/-80, 1460 +/-30, 1465+/-35, 1660+/- 30, 1830+/-15, 1875+/-10 and 1910+/-10 CE. Age-depth modeling shows increased sediment accumulation rates following ca. 1800 CE. Particle-size analysis of historic floods demonstrates a relative increase in fine sand (RFS) content with increasing flood magnitude, which is consistent with previous work upstream. The highest RFS values are found within flood deposits dated to ca. 1830 CE, 1460+/-30 and 1875 +/-10 CE, the latter of which coincides with the 1867 CE historic flood along the Tennessee River. The high magnitude flood of 1830+/- 15 CE is consistent with USGS paleoflood analysis upstream that documents a paleoflood occurring ca. 1600-1800 CE that was higher in elevation than the historic flood of record. The earliest observed flood deposits appear to occur during transition into the Medieval Climate Anomaly between 800 and 1300 CE with increased flood magnitude through the Little Ice Age (1400-1800 CE), and with peak magnitude occurring 1830+/-15 CE.

The dispersal of Homo sapiens across and out of Africa during the Late Pleistocene (130 –10 ka) is a major event in the history of our species. These important events are recorded within sedimentary deposits of East African basins. The Late Pleistocene site of Farre is located within the Chalbi basin of northern Kenya, and is an open-air site containing thousands of artifacts and fossils. This study attempts to reconstruct the depositional and weathering environments of Farre through comparison with modern analogues in the Chalbi basin that include proximal, medial and distal alluvial fan settings, dunes and playa.
Today, the Farre site is exposed on an interdune setting along a proximal alluvial fan. Optically-stimulated luminescence (OSL) ages from Farre site sediments show that the site is likely older than 56 ka, which exceeds the maximum age for OSL dates on quartz in the basin. The the overlying sediment and soil are ~22 ka. The nearby Menengai Tuff (36 ka) is at a similar elevation with respect to Farre and is found in comparable stratigraphy and provides support that the site has a Late Pleistocene component. The ratio TiO2/ZrO2, an indicator of parent material uniformity and provenance, was compared between Farre and the modern-analogue sites. Farre has a mean and standard deviation TiO2/ZrO2 of 12.11±1.39, which is most similar to the modern quartz-rich distal fan analogue, 11.79% ± 2.03. The Chemical Proxy of Alteration (CPA) and Weathering Index (WI), were also calculated for Farre and each modern-analogue. The mean CPA and WI values of the artifact-bearing layers at Farre are 77.72% and 9.28% respectively, similar to modern medial fan soils, having values of 74.25% and 12.57%. Although Farre sediment is similar to that found on a quartz-rich distal fan, the CPA and WI data suggest that the weathering pathways differ.

INVESTIGATION OF OMNIVOROUS TROPHIC POSITION IN THE DRILLING GASTROPOD, UROSALPINX CINEREA, USING STABLE ISOTOPE ANALYSIS
Drill-holes found in the fossil record are an important tool to study ecological patterns of the past. It is therefore important to gain a better understanding of the role of extant drilling snails in modern ecosystems. Although traditionally considered a predator, trophic position of 3.0, specimens of the muricid Urosalpinx cinerea from Long Island Sound revealed trophic positions between 2.3 and 2.5, suggestive of an omnivorous diet. This study addresses the generality of this result by examining a U. cinerea population from Wilmington, North Carolina. Preliminary whole body, soft tissue stable isotope analysis of nitrogen and carbon was conducted on five U. cinerea specimens. Isotopic baseline for the study area was calculated using proxy taxa, including Geukensia demissa for the pelagic baseline and Littoraria irrorata for the littoral baseline. Trophic position for these U. cinerea specimens ranged from 2.4 to 2.9. Working hypotheses to explain a trophic position lower than 3.0 in U. cinerea include: trophic omnivory driven by plant consumption, or a lower-than-average nitrogen discrimination factor. Although no studies on the nitrogen fractionation factors of muricids currently exist, the naticid Neverita duplicata from Long Island Sound has recently been demonstrated to have a normal nitrogen fractionation factor and omnivorous isotopic signatures. The difference between the trophic ranges of the two locations may indicate that U. cinerea have a more predatory diet in North Carolina than in Long Island Sound. However, further work is needed to confirm that these values reflect dietary differences, not a below average nitrogen fractionation factor.

In the event of a spinal cord injury (SCI), individuals are often subject to skeletal muscle deterioration and adipose tissue gain in paralyzed muscles. These negative impacts can limit motor functions and lead to secondary complications. A novel framework was proposed to automatically segment the MRI volumes of human thigh into fat and muscle volumes. Additionally, within the muscle volume, three main muscle compartments: quadriceps femoris, hamstrings and adductors were segmented. The proposed method for fat segmentation has been shown previously to have a high accuracy (95.51±1.61). Further, muscle group segmentation has too been shown to have high accuracy (94.76±1.70). The accuracy measures were based on dice similarity percentage calculated on 10 spinal cord injured (SCI) and 10 non-disabled (ND) individuals.

Demographic, occupational, tumor characteristics and extent of metastasis of metastatic melanoma patients diagnosed in Florida, 1996-2010.


Recent decades have witnessed an increase in melanoma more than any other cancer, resulting in a 2010-2014 age-adjusted incidence rate (IR) of 22.3 and 2.7 per 100,000, respectively. Florida's IR is the 2nd highest in the nation and mortality rate has doubled since 1975. Although metastatic melanoma (MM) is less frequent among minorities, it has been increasing steadily over the years. AIM: To describe the demographic, occupational, tumor characteristics and extent of metastasis of MM patients. A dataset of 80,349 Whites, African Americans (AA) and Hispanic stage II and IV MM patients at presentation was obtained from Florida Cancer Registry. Demographic information, occupational status and measures related to age at diagnosis, primary site and laterality, histology, and grade and stage of the cancers are reported. Data were analyzed using SAS. Means ± SD, frequencies and percentages, and chi-square tests were employed at P <0.05. Fifteen counties out of 67 accounted for 72% of all MM cases; 61% of the patients were married at the time of diagnosis. Majority reported having government sponsored coverage and sustained by outdoor occupations, while 60% had never smoked. Sixty-nine percent of patients were diagnosed with tumors of the trunk, shoulder and hip; laterality was evenly distributed between left 39% and right 37%. More African Americans and Hispanics had tumors that were either moderately or poorly differentiated. This study confirms the well-established race, gender and age disparity in MM diagnosis – majority white and male, and majority of the cases between 56 and 71 years of age.

In this preliminary study, we hypothesized that ingestion of phytochemicals would significantly alter salivary and urinary pH. Salivary and urinary pH reflect dietary systemic net acid load in the body. Numerous metabolic and degenerative diseases have been implicated in the list of health compromises that accompany acidic load dietary intake. There is considerable debate in the lay press, as well as the scientific literature, about the health benefits of an alkaline diet. This preliminary study investigates whether pH changes can be reliably measured acutely (within 30 minutes) upon ingestion of a relatively moderate alkaline phytochemical. The aim of this study is to provide a basis from which to ask more pointed physiological questions about the effects of acid load on other organ systems of the body.

Bighead and silver carp, commonly called Asian carp, are non-native fish that have negatively impacted North American waters including Kentucky Rivers and lakes. Harvesting Asian carp for human consumption has been proposed as one of the tools to reduce or eliminate Asian carp from Kentucky waters. Intramuscular bones in Asian carp has to be removed in order to attract American consumers. However, the process of mechanical deboning Asian carp destroys the structure of the fish muscle, limiting the options of products forms that can be made from Asian carp meat. Salt can solubilize muscle proteins and increase binding. The objective of this project was to determine optimum of salt concentration to increase binding in fish meat. Asian carp captured from Mississippi River were deboned and ground through 5mm screen by a commercial fish processor. Samples of Asian carp mince were mixed with salt at the concentration of 1%, 2%, 4%, 6%, respectively, and then blended for 30 minutes with a Blender (Kitchen Aid KSM75WH) with the speed setting at 4. Meatballs were formed manually, and heated in hot water at 80°C for 8 minutes. The hardness and stickiness of fish meatball were measured with Texture Analyzer (TX2 Plus). The hardness of meatball was increased from 370g to 630g by salt of 1% to 6%, meanwhile the stickiness also increased from 15g to 23g. At salt of 4%, the hardness is already 576g. The results suggest that 4% of salt may be needed to generate the necessary binding needed to form Asian carp meat meatball.

Pancreatic cancer is the 4th leading cause of death from cancer and its incidence has risen steadily. The survival rate of pancreatic cancer patients remains the lowest among all cancer types, and is projected to increase further in the future.
Septins family proteins bind to GTP of G protein, which allow to form a stable complex by interacting between a choice of members of the family. Structurally, Septins are related to RAS oncogene, and has been shown as an oncogene by its association with the mixed-lineage leukemia gene. Septin expressions are altered in many cancers, and their mutations were reported in cancers of large intestine, skin, endometrium and stomach. However, the expression and function of Septins in pancreatic cancer has not been examined.
Our preliminary data on PCR demonstrates increased level of transcripts of Septins in pancreatic adenocarcinomas compared to normal pancreatic cells. Our data on Western blot and imunocytochemistry indicates that protein level of various Septins are increased in pancreatic adenocarcinomas compared to normal pancreatic cells. Immunohistochemical analyses also shows higher Septin protein expression in cancer cells or cancer tissues each compared to normal cells or tissues.
We investigated the function of Septins by generating stable clones of Septin knockout using CRISPR approach in pancreatic cancer cells derived from malignant adenocarcinomas. Knockout efficiency was confirmed by GFP expression and Western blot analysis. Compared to control knockout, Septin knocktout induced a significant growth inhibition in cancer cells.
In summary, our preliminary data indicates that Septins promote the development of pancreatic cancer by increasing their transcripts and proteins. Targeting Septin members may be a potential approach for the development of new therapeutics of pancreatic cancer.

The purpose of this pilot study was twofold: 1) to collect trail user type, frequency, and duration on two separate multi use paths in a rural Appalachian city, and 2) to estimate the health impact of the trails. In addition, this study served to establish baseline measures prior to the completion of an addition shared use path which will connect the segments. Observational data, intercept user surveys, along with infrared sensors were used to estimate the number and type of users. Data were collected for 12 hours (6:00 am-6:00 pm) on two weekdays in July of 2017. Data from surveys were analyzed in tandem with information from the counts to develop an estimate of annual visits. Weather data (precipitation and temperature) from previous research was used to create relative ratios of use for each day of the year. The health impact model used the relationship between exposures to trail use, verses no use, to estimate all-cause mortality using the model established by Götschi & Hadden Loh (2017). The results indicated that the approximate number of annual users (primarily walkers) for both trails combined was 7,784. Out of this number of individuals, the number who would be expected to die if they were not walking regularly would be 71. In addition, the number of deaths per year that may be prevented by this level of walking was 13. Future data collection will provide annual usage, frequency and duration as the multi-use path is connected and therefore lengthened.

Hepatitis C virus (HCV) is one of the most prevalent and deadly blood-borne pathogens. No vaccines are currently available to prevent or treat HCV infection. Using recombinant DNA techniques, we developed several novel vaccines for HCV using a modified herpes simplex virus backbone. Mice were immunized with vaccine DNA alone or vaccine DNA packaged in herpesvirus particles. Antibody responses were measured by ELISA following vaccination. Most vaccine groups developed greater than 10 ug/ml of HCV-specific antibody, demonstrating that all of the vaccines were immunogenic and stimulated high levels of serum immunoglobulin G (IgG). We are currently measuring T cell responses in the various vaccine groups. These results demonstrate the immunogenicity of novel HCV vaccines in a mouse model. These vaccines may someday be used to prevent HCV infection in man.

Measurement of Influenza Severity in Tennessee Hospitalized Patients, 2016-2017. Allison Harper1,2; Tiffanie Markus, PhD, CCAP1; Gail Hughett, RN, BSN, CCRP1; Danielle Ndi, MPH1; Karen Leib, RN, CCRP1; H. Keipp Talbot, MD, MPH1. Vanderbilt University Medical Center, Departments of Medicine and Health Policy1; Berea College2.

Vanderbilt University Medical Center's (VUMC) Emerging Infections Program (EIP), as well as the Tennessee Department of Health, collaborates to report yearly seasonal Flu cases to the Centers for Disease Control and Prevention (CDC). Previous studies that have been conducted among the EIP Influenza sites have recognized the positive correlation between clinician testing and rates of hospitalization observed by sites and the fact that testing varies from site to site. Therefore, testing for influenza is often underutilized due to poor reliability of rapid test results and/or greater reliance on clinical diagnosis for influenza (CDC 2016). We hypothesis this experiment will collect data on the chosen variables of the hospitalized patient that may be used to generate an influenza severity score. The influenza severity score would be able to aid in the community and hospital's process that will enable the admission of the most severely ill patients efficiently antecedently the other patients. The experimental design is abstracting data from the electronic data records, scribing the selected variables on Redcap for each patient, and analyzes the results using statistical software (SAS). Subsequently, the measurements of the processes for collecting the feasibility of the obtained patient's variables, and the time efficiency for the chart reviewer to complete each case were evaluated for this experiment. The experimental pilot that only takes into account a subset of the 2016-2017 flu population, therefore no true correlations can be created.

Objective: The purpose of this study was to investigate the positive and negative psychological adaptations due to of secondary traumatic stress and the role of resilience among paramedics and Emergency Medical Technicians.

Methods: Pre-hospital Emergency Medical Service (EMS) providers from multiple ambulance dispatch centers at a Regional Health System anonymously completed four validated questionnaires: Secondary Traumatic Stress, Posttraumatic Growth, Resilience, and Changes in Outlook. Relationships between these constructs, perceived level of symptoms, and demographics were explored.

Results: Data from 53 EMS providers were analyzed. Overall, a moderate degree of resilience, secondary traumatic stress, and negative change in outlook were observed with a higher than average positive change in outlook. Resilience was significantly (p <0.001) inversely related to secondary traumatic stress and negative change in outlook. EMS working part time demonstrated a significantly higher (p=0.005) level of resilience compared to those working full time. Likewise, post-traumatic growth was significantly (p=0.03) higher in EMT's while resilience was significantly (p=0.02) higher in paramedics. Additionally, resilience and growth scores among those who expressed interest on receiving coping training was significantly higher (p <0.02) than respondents who were not interested to receive similar training. No significant differences (p>0.05) were detected between years of experience for any attributes analyzed.

Conclusions: The findings demonstrated significant correlations between secondary traumatic stress, resilience, growth, and changes in outlook in EMT's and paramedics.

Background: Physical activity during pregnancy has benefits for pregnant women and their offspring. Studies suggest that exercise during pregnancy may also elicit improvements in the brain of the newborn. To our knowledge, influence of physical activity on infant motor development has not been studied.

Purpose: Therefore, the goal of this project is to determine the relationship between maternal physical activity during late pregnancy and infant motor development.

Methods: Physical activity during late pregnancy (32-39 weeks) was assessed via Actigraph Link accelerometers. Infant motor development will be assessed using the Alberta Infant Motor Scale (AIMS) in four month old infants. The AIMS is a validated test to assess motor control in infants from birth to 18 months of age, and all testing will be performed by a physical therapist who is board certified in pediatrics. Maternal physical activity and infant AIMS scores will be correlated using SPSS in order to determine the relationship between exercise during pregnancy and newborn motor control/development. In addition, potential confounders (e.g. amount of time infant spends in different positions, number of siblings, daycare environment, etc.) will be assessed via surveys our team has developed.

Expected Results and Significance: We believe the maternal physical activity levels during pregnancy and the AIMS scores in infants will be positively correlated. These findings may suggest maternal exercise as an effective and resourceful approach to improving maternal and infant health, as motor performance early in life is linked to improvements in cognitive function in childhood.

Chronic diseases such as diabetes, cancer, atherosclerosis, stroke, and cardiovascular diseases are often a result of oxidative stress. Oxidative stress occurs due to an imbalance between the rates of generation of Reactive Oxygen Species (ROS) and clearance by the antioxidant mechanisms in the body, sometimes leading to interaction with biomolecules such as proteins and DNA. One of the critical metabolite of oxidative stress is 8-hydroxy-2'-deoxo-guanosine (8-OHdG), which is a DNA adduct. Higher levels of which can escape the DNA repair and result in point mutations. This study attempted to evaluate use of 8-OHdG levels in spot urine samples in young adults with and without Metabolic Syndrome (MetS) as a non-invasive method. First year college students (n=376), average age 19.8 years), attending Kentucky State University, Frankfort, with no prior diagnosis of illness participated in the cross sectional study. Anthropometric screenings included measurement of height, weight, waist circumference, and body mass index. The clinical screenings included measurement of blood pressure and determination of fasting lipid and glucose concentrations. The National Cholesterol Education Program's Adult Treatment Panel III (NCEP ATP III) and International Diabetes Federation (IDF) definitions for MetS were applied. 8-OHdG concentration levels were measured by Enzyme Linked Immunosorbent Assay (ELISA). Analysis of variance (ANOVA) scores on the Means were used to examine differences between genders for all parameters. Correlation of Means (p ≤ 0.05) was used to determine significant correlation. Prevalence rate of MetS based on NCEP ATP and IDF definitions were found to be 12% and 9% respectively. Of the various Mets criteria, only high blood pressure correlated with higher urinary levels of 8-OHdG. The study indicates the possibility of using 8-OHdG as a marker of oxidative stress, however, analysis using a 24-hour urine sample is desirable.

Caleb Sedlak (a) ,Minsub Chung(b) , Moon-Soo Kim(a)
(a) Department of Chemistry, Western Kentucky University,1906 College Heights Blvd, Bowling Green, KY 42101 USA
(b) Department of Chemical Engineering, Hongik University Seoul, Korea
Zinc Finger Proteins (ZFPs) are one of the most common DNA-binding domains. ZFPs can be engineered to bind to specific genes on a double-stranded DNA. Developing a rapid and reliable method for detecting specific pathogens would be greatly beneficial to modern biomedicine as well as more resource-limiting areas. A pair of ZFPs was used in a two-step process to first capture the target DNA and then apply the second detection probe ZFP labeled with a fluorescent molecule. A stx2 gene was chosen as a target DNA, which encodes for Shiga toxin a foodborne pathogen E.coli O157. The ZFP array takes the capture ZFP probe and immobilizes it on an acrylamide gel surface. After target DNA was added, the detection probe a ZFP labeled with a fluoresces molecule was applied to the bound complex of the capture probe and target DNA. At the final step, fluorescence intensity was measured to compare the signals between target DNA and non-target DNA.
Part of this research was carried out through the collaboration between WKU and Hongik University in South Korea through the National Science Foundation IRES Program. This collaboration helps to gain advanced research experience, and as allowed an opportunity to learn and experience Korean culture.

Approximately 1 out of every 68 children in the US are diagnosed with Autism Spectrum Disorder (ASD). ASD is a heterogeneous neurodevelopmental disorder that has widespread activation of brain immune cells that produce inflammation. Data suggest diet can be used to control inflammatory effects in other conditions, such as diabetes, which is regulated by the anti-inflammatory characteristics of zinc. Our aim is to assess whether manipulation of dietary zinc and fat have interacting behavioral and physiological effects on mice. In this study, an inbred polygenic mouse model of autism (BTBR) was used. Mice (54 M, 61 F) were separated into 6 groups, based on dietary zinc (low, normal, high), and fat (normal, high) content. Behavior was analyzed using open field, 3-chamber, and an elevated +-maze tests, recorded from the side and/or above, and analyzed using in-house machine vision software. Only open-field data are presented here. Data were collated in excel and analyzed via multi-level ANOVA in R.

This study addressed the inactivation of fungal spores from three different genera (Fusarium, Penicillium, and Mucor) with temperatures from 50°C to 80°C or UVC doses of 50 to 500 millijoules (mJ) per cm^2. Fungal spores were prepared by washing growing fungal colonies with a saline solution containing 0.05% Tween20. Spore suspensions of approximately 1,000,000 (10^6) spores per milliliter were exposed to heat or UVC and then diluted and plated on sabouraud dextrose agar (SDA). Plates were examined for germination and growth for seven days. The results demonstrated clear evidence that heat was an effective way of inactivating fungal spores from these three genera. Fusarium and Penicillium spores were killed by thirty minute incubation at temperatures >50°C, while Mucor spores died after thirty minutes at temperatures >65°C. Exposure to low doses of UVC (100 mJ/cm^2) reduced spore viability of all fungi by >99% (>2log10), and 200 mJ/cm^2 reduced spore viability below the limits of detection (>4log10). In general, Mucor spores were the most resistant to heat, but appeared to be most sensitive to UVC. The procedures and knowledge learned from this project will benefit future research pertaining to medically and environmentally relevant fungi such as those linked to White Nose Syndrome, a fungal disease that is killing bat populations in North America.

Mycobacteriophages are viruses that infect bacterium in the genus Mycobacterium and are readily found in environmental samples. The Science Education Alliance-Phage Hunters Advancing Genomics and Evolutionary Science (SEA-PHAGES) is a mycobacteriophage research project that has two distinct portions: phage isolation and phage genome annotation. In the spring of 2016, two mycobacteriophages were discovered from various soil samples in North Central Kentucky using Mycobacterium smegmatis as host. These two mycobacteriophages, Spock and Inky, were difficult to amplify and titer but were sufficiently amplified for DNA extraction at Western Kentucky University. Their genomes were sequenced at North Carolina State University and both Spock and Inky were determined to belong to subcluster K1. These genomes are being annotated this fall using the Phage Evidence Collection and Annotation Network (PECAAN). Phages in subcluster K1 typically have a 11 bp 3' overhang. Preliminary results suggest that the Spock and Inky genomes encodes 96 and 95 gene respectively and no encoded tRNA. The K1 subcluster is of particular note because most of the mycobacteriophages in this cluster can infect Mycobacterium tuberculosis . This research will add to the growing knowledge of mycobacteriophages, with the ultimate goal of discovering new lab techniques and treatments for the pathogen M. tuberculosis.

Endophytes, microorganisms that grow within plant tissues without causing disease, often produce chemical compounds which benfit their host by fostering growth or killing pathogenic microbes. These compounds may be clinically useful in treating infections in humans. In this study, fungal endophytes were collected from plants in the Northern Kentucy area and were tested for antimicrobial activity using co-culture assays. The eleven isolates that showed activity were identified through DNA sequencing and BLAST analysis. The compounds responsible for inhibiton were extracted from liquid cultures for further testing and verification of anti-fungal activity. Overall, the diversity and abundance of antimicrobial endophytes offers promising new direction in the search for sources of antibiotics.

Several mycoplasmas have been previously found to possess a series of pathways used for production of hydrogen peroxide (H2O2) which may be used as a virulence factor to damage host tissues. Two potential pathways for H2O2 production are present in the genome of Mycoplasma mycoides subspecies capri (Mmc) and were further investigated in this study. Strains of Mmc lacking one or more proteins involved in H2O2 production were analyzed using H2O2-production assays with a constant concentration of bacterial cells. Assays were incubated at room temperature in the presence of glycerol, glycerophosphocoline (GPC), or no sugar, at which time H2O2 production was measured with colorimetric test strips. The survival of Caenorhabditis elegans nematodes co-incubated with Mmc was also measured to determine the effect of H2O2 production on virulence. The addition of either glycerol or GPC increased H2O2 production and decreased C. elegans survival for most strains tested as compared to the no sugar control. Strains of Mmc lacking multiple proteins typically displayed the lowest levels of H2O2 production and highest survival rates of C. elegans. The removal of the protein GlpO from Mmc had the greatest singular elimination effect on the production of H2O2 and the percent survival of C. elegans. Combining the removal of GlpO with the elimination of other proteins from the production pathway has the greatest overall effect. Therefore, it appears that GlpO is the most influential protein in the H2O2 production pathway.

Antibiotic resistant bacteria have been identified by the World Health Organization and the Centers for Disease Control and Prevention as a global threat. Because they are exposed to frequent inputs and constant flux of various components, aquatic environments make effective reservoirs for a variety of microbes. Such sites are often in contact with humans and may provide sources of infection if pathogenic microbes are present. Contamination of waters with antibiotics may select for the maintenance of antibiotic resistant bacteria in the communities present there.
To identify the risk associated with antibiotic resistant bacteria in the waters around Louisville, KY, microbial communities in soil and water samples associated with wastewaters, natural waters, agricultural waters, and public use waters (i.e. drains and faucets in public places), were tested to determine whether they contained bacteria able to grow in the presence of colistin and vancomycin, two antibiotics of last resort
Individual antibiotic-resistant bacterial strains were isolated and identified through Sanger sequencing of the 16S rRNA gene. The variety of genera and species identified supports our hypothesis that these environments are rich in resistant bacteria. This data, though subject to the limitations of culture-based diversity approaches, allows us to compare types of aquatic environments. The isolated strains will be further characterized for their potential to cause disease, and eight strains have been chosen based on their clinical significance and/or rarity for full genomic sequencing.

Bacteriophages are viruses that infect bacteria and are found in many different environments. A growing body of data are being acquired to understand these viruses. As part of the Kentucky Biomedical Infrastructure Network Small Genomes Discovery Program at Somerset Community College, the mycobacteriophage Aquire49 was isolated and purified from soil samples collected in London, KY. The general plaque morphology showed clear plaques that were 6.54 mm2 in diameter indicating that the phage was likely a lytic phage. Based on Transmission Electron Micrographs the phage had an average capsid length of 56.53 nm and an average tail length of 249.06 nm. The genome was sequenced using DNA extracted from the purified phage lysate of Acquire49. Annotation of the 73,639 bp Aquire49 genome was performed using PECAAN (Phage Evidence Collection and Annotation Network, which resulted in 121 putatively annotated genes. Blastn analysis matched Acquire49 with members of the L1 subcluster in the Actinobacteriophage database.

New antimicrobial drugs are needed to treat the growing number of drug-resistant bacterial infections and invasive fungal infections. The goals of this project are to isolate and identify antimicrobial-producing microbes found in the environment and to extract and identify the active compounds they produce. Environmental samples were collected from the Northern Kentucky/Cincinnati area. Isolates were tested for activity against bacterial (Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Klebsiella pneumoniae) and fungal organisms pathogens (Candida albicans, Candida glabrata, Candida krusei, and Candida tropicalis) and active isolates were identified using 16s rRNA gene sequencing. Liquid cultures of active isolates were extracted with Amberlite XAD16N and the resulting extracts were tested for activity using well- or disk-diffusion assays. These compounds still produced some antifungal or antibacterial activity. Efforts are currently underway to purify and elucidate the structure of active compounds for assessment as potential leads in new drug development.

Infection due to antibiotic resistant bacteria is a significant problem for clinicians and patients. Due in part to improper use of antibacterial agents, resistance in microbial populations in the environment, a potential source of such infections, is increasing.
To assess the diversity of microbes found in aquatic environments around Louisville, KY, whole communities of microbes as well as the subset resistant to colistin and vancomycin, two last line antibiotics, were isolated from 48 water sources classified in four categories: agricultural waters, natural waterways, wastewaters, and sink drains in commercial establishments. 16S rRNA libraries were generated from each sample, sequenced using high throughput sequencing, and OTUs, alpha diversity, and beta diversity were assessed.
The most common genus of bacteria, Ochrobactrum – a genus containing several opportunistic pathogens, was found in 78 out of 87 samples with an average abundance of 31% across all samples but appeared to be enriched in antibiotic-treated samples (average 42% vs 19% in control samples). Other common isolates belonged to the genera Microbacterium, Leucobacter, and Serratia. In general, agricultural samples showed a greater diversity than drains, natural water, or wastewater samples, a finding that we hypothesize may be due to increased carbon inputs from plants and animals or carbon-rich drainage. Additionally, there were significant differences in alpha and beta diversity between sites or isolation conditions. Understanding the natural reservoir of antibiotic resistance will lay the groundwork for understanding emerging antibiotic resistant pathogens and possibly inform new policies dictating the usage of antibiotics for non-therapeutic uses.

Natural transformation is a widespread mechanism of horizontal gene transfer in bacteria and a major driver of evolution which promotes the increasing prevalence of antibiotic resistance. The obligate human pathogen, Neisseria gonorrhoeae, undergoes natural transformation frequently which has led to antibiotic resistant strains which fail clinical treatment. Several related non-pathogenic Neisseria species reside in the human nasopharynx, contribute to the normal human microbiome, and are likely genetically linked to N. gonorrhoeae. Although much research has focused on the pathogenic species, little is known regarding the genetics of the commensal (non-pathogenic) Neisseria species. Here, we conduct Minimum Inhibitory Concentration (MIC) assays, isolate two resistant mutants, and demonstrate transformation in Neisseria sicca ATCC 29256, a sequenced commensal strain previously thought to be refractory to transformation. Importantly, our work has established protocols and reagents needed for further study of N. sicca and supports the notion that genetic exchange between the pathogenic and non-pathogenic Neisseria occurs in the nasopharynx. This inter-species genetic transfer likely contributes to the evolution of virulence determinants and resistance to antibiotics.

The Toxic Effects of Chemically Synthesized and Biologically Fabricated Gold Nanoparticles on Caenorhabditis elegans Fitness

The ever-increasing application of metal nanoparticles in consumer products has raised concerns about the environmental safety of nanoscale materials. In order to obtain a more complete understanding of the toxic effects nanomaterials present, this study was conducted to examine the possible life cycle and behavioral changes these materials induce in the model organism Caenorhabditis elegans. The effects of gold nanoparticles (AuNPs) of two origins (biologically fabricated in plants and chemically synthesized commercial products) were examined at concentrations of 30 and 50 ppm over a three-day period. Exposure experiments indicated reductions in the fitness of adult populations as evidenced by lower levels of progeny. This study compared the overall effects of biologically fabricated AuNPs with those of chemically synthesized AuNPs and found that the two materials have similar toxic effects on C. elegans fitness. The data collected provides a more complete knowledge of the toxicity associated with AuNPs on a living system. Genetic analyses of certain target genes are currently underway in hopes of determining any genomic alterations that could be inducing reductions in C. elegans fitness.

Amber Earlywine1,2, Tyler Kinder3, Dr. Rebecca Dutch1,3 1NSF REU program, Lexington, KY; 2Berea College Department of Chemistry, Berea, KY; 3Department of Cellular and Molecular Biochemistry, University of Kentucky Lexington, KY; The fusion protein (F protein) has shown to be the only surface protein necessary for viral transmission of the negative sense, ssRNA virus, Human Metapneumovirus (HMPV). The F protein is activated through cleavage by proteolytic activity. The cleavage event causes a conformational change in the F protein where Heptad Repeat A (HRA) moves from the globular head, attaches to the membrane of the target cell, and moves to the stalk domain, fusing the two membranes. Based on previous knowledge of how serine proteases cleave Hemagglutinin (HA) in Respiratory Syncytial Virus (RSV), it is believed that these proteases also cleave the Fusion protein in HMPV. The goal of this project was to test if different proteases such as HAT (TMPRSS11D), TMPRSS2, TMPRSS4, Matriptase, KLK5, or KLK12 cleave the F protein so that we can use a protease inhibitor and stop viral spread. However, there was a focus on TMPRSS2 and TMPRSS4. First, expression of TMPRSS4 was tested using Western Blot Analysis, using HA as a positive control. When no expression was seen, TMPRSS4 was sub-cloned into pcAGGS from pcDNA and cleavage was not seen. TMPRSS2 expression was also tested through Western Blot Analysis with trypsin as a positive control and cleavage of the F protein was observed, suggesting that the TMPRSS2 protease can be inhibited.

RR Lyrae (RRL) stars are well known standard candles that can be used to map out the structure of the Milky Way galaxy. In order to use these stars, however, one needs to be able to identify them effectively. One major issue in doing this is determining the pulsational period of an RRL star. In this poster, we will analyze different period finding techniques on RRL from the Kilodegree Extremely Little Telescope (KELT). In particular, we will focus on trying to separate RRL stars from binary stars, which can be easily mistaken for RRL stars if the wrong period is found.

How electromagnetic waves are affected by the geometry around a black hole can tell us important information about the black hole itself. By taking different variables that affect the geometry of the space around it, and how light is affected by said geometry or the potential of the black hole, we get the frequency and damping of the wave after interacting with the black hole, shown as ωr and ωi. We can calculate these frequencies for different values of mass and charge. Also, we can see how temperature is related to the frequencies of the electromagnetic waves. The properties of the black hole determine how it will affect the electromagnetic waves, so by understanding how each property affects them, we can see how changes in the electromagnetic waves indicate the specific properties of the black hole.

In this poster, we will use the color, period, and amplitude of stars in the Kilodegree Extremely Little Telescope (KELT) survey to identify RR Lyrae (RRL) stars. RRL stars are important because they can be used as tracers of galactic structure. The KELT survey is a single filter survey with between 4,000 and 9,000 epochs. Although this large number of epochs is useful for defining the light curve shape, it is difficult to separate some types of RRL stars from eclipsing binary stars by shape alone. To supplement these light curves, we will use catalog magnitudes to find color relations that can separate the RRL from the eclipsing binaries. We will then use the combination of these relations with light curve shape in order to pick out RRL stars

Graphene-mediated surface-enhanced Raman scattering (G-SERS) is a recent phenomenon that produces clean and reproducible Raman signals of chemical molecules with significantly enhanced intensity. Since G-SERS relies on a chemical mechanism and therefore it shows molecular sensitivity and selectivity. We developed graphene-family nanomaterials, GFNs, decorated with coinage silver and gold nanoparticles for detection of methylene blue (MB) and rhodamine 6G (Rh6G) probes in view of optical and biological importance. The results illustrate that silver and gold nanoparticles immobilized on GFNs enhanced the Raman signal, in general, and as cascaded amplification of on multilayer architecture, larger than those on the metal nanoparticles without graphene. Additionally, the sensitivity can be tuned by controlling the size of nanoparticles. Moreover, highly-sensitive graphene-nanoparticle sensors are capable of molecular detection over 10 pM to 100 microM concentration. The G-SERS enhancement is discussed in terms of graphene-metal nanoparticle interactions leading to local interfacial hybridization and polarization, 2. molecular conformation of analyte on nanoparticle-graphene functionalities, and 3. charge transfer and exchange or sharing of charges between analyte and nanoparticles decorated graphene supports, experiencing chemical enhancement. Optimized metal nanoparticle-graphene electronic properties are determined from density functional theory (DFT) calculations.

In this work, we developed large-area nanofiltration membranes using 1) shear aligned discotic nematic phase of graphene oxide and 2) holey graphenes with narrow hole size distribution via controlled catalytic oxidation. We also prepared interconnected network of mesoporous graphene-based electrodes to achieve optimal desalination during capacitive deionization (CDI) of brackish water, attributed to higher specific surface area, electrical conductivity, good wettability of water, environmentally safe, efficient pathways for ion and electron transportation, as potential successor of current filtration membranes. The pressure driven transport data on highly ordered, continuous, thin films of multi-layered graphene oxide and holey graphene is expected to demonstrate faster transport for salt water, higher retention for charged and uncharged organic probe molecules with hydrated radii above 5Å as well as modest retention of mono- and di-valent salts for ~150 nm thick membranes. The highly ordered graphene nanosheets and nanoscaled porous graphene in the plane of the membrane make organized, molecule-hugging cylindrical and spherical channels, respectively, thus enhance the permeability and hydrodynamic conductivity. The results illustrate that both the macro and nanoscale pores are favorable for enhancing CDI performance by buffering ions to reduce the diffusion distance from external electrolyte to the interior surfaces and enlarging surface area.

Nanoparticles-grafted functionalized graphene coated with nanostructured polyaniline layered nanocomposites as high-performance biosensors.
The challenge remains to develop (chemical, electrochemical and biological) sensors from nanocomposites with broader electrical conductivity, molecular sensitivity and specificity. We report the design and synthesis of scalable, metal nanoparticles-grafted functionalized graphene overcoat with nanostructured polyaniline nanocomposites and elucidate their high-performance as advanced biosensors. The versatility of the nanocomposite performance was corroborated by altering the size, areal density and morphology of electrodeposited gold (AuNPs) and silver (AgNPs) nanoparticles on the nitrogenated functionalized graphene (NFG) as well as the density of electropolymerized polyaniline (PANi) onto NFG. Gold and silver NPs are selected due to their higher electrical conductivity, facile synthesis, easier processability and scalability. The critical modification of architectures (NFG/AgNP or AuNP/PANi) on FTO electrodes increased the conductivity of the electrodes significantly and reduced the charge transfer resistance dramatically while investigating electrochemical properties. The high-performance biosensing application is demonstrated for the detection of ascorbic acid (AA) over electroactive components interfering species commonly found in blood serum samples, with enhanced sensitivity over a range of detection thereby determining limit of detection. These nanocomposites are applicable for electrocatalysis, energy systems as well as enriching biofuel cell development.

Surface Functionality Of Graphene-family Nanomaterials On Electronic And Phononic Properties For Applied Electrochemistry.
We present optical and lattice vibrational properties of a range of functionalized graphene-family nanomaterials using UV-visible absorption, photoluminescence excitation (PLE) and micro-Raman spectroscopy techniques. Various functionalized graphene nanomaterials include few layer graphene oxide, reduced graphene oxide, graphene quantum dots and three-dimensional scaffolds graphene aerogel and their nitrogenated functionalized counterparts. Raman spectroscopy (RS) provides lattice dynamical structural characterization at nanoscale revealing collective atomic/molecular motions and localized vibrations. The role of oxygen epoxy (C-O-C, carbonyl, C=O) and nitrogen (pryridinic and graphitic/pyrrolic) functionalities and corresponding bonding configurations with quantum size effects are emphasized in view of understanding physico-chemical properties for biosensing and water desalination. While first- and second-order phonon modes are analyzed in terms of Raman intensity, band position (intrinsic mechanical strain) and intensity ratio (structural disorder, number defect density), distinct localized π electronic states were found in PLE spectra at the carbon atoms around oxygenated and nitrogenated species. The origin of these states is discussed based on experimental findings and density functional theory exemplifying structural evolution.

We have incorporated commercially produced carbon nanotubes (CNTs) into GexSe1-x glass system with different x values (21.5%, 22.5% and 23.5%) and investigated the effect of incorporation of CNTs on glass transition temperature. Our GexSe1-x glass system was successfully synthesized with x = 0.225 and we have observed it undergo a transition to a new stress-free phase between 225o C and 240o C with advanced material properties where x was between 0.215 and 0.235. Modulated Differential Scanning Calorimetry study indicates that the glass system with the incorporation of CNTs undergoes the transition at a different temperature compared to the glass system without CNTs. In order to make sure that the change in glass transition temperature is due to the exotic properties of CNTs, we have also studied the effect of incorporation of pure carbon into GexSe1-x glass system with different x values (21.5%, 22.5% and 23.5%). This study is important because it incorporates the exotic properties of CNTs and the stress-free phase of the GexSe1-x glass, which may lead to a system that could potentially have advanced material properties.

The study of bending light paths around black holes
Abby Campbell and Dr. Sharmanthie Fernando
During this project I studied how the geometry around black holes affects the objects, specifically photons, that orbits around them. I first gained a basic grasp of knowledge about Einstein and his theory of general relativity. I was also given literature describing what a black hole is and how it relates to space-time. I studied the equations which governs the behavior of photons around a black hole. I calculated the path for photons for three energy levels. These energy levels were determined based on the effective potential corresponding to the photon motion. Mathematica software was used extensively to plot the graphs and the path of the photons.

A commonly used point-of-care (POC) method to detect hemoglobin (Hb) uses Vanzetti's azide-methemoglobin method (HemoCue analyzer). Since azide is explosive and toxic, this research was aimed to develop a greener/safer assay that utilizes the indicator 3, 3', 5, 5'-tetramethylbenzidine (TMB), which donates electrons to oxidized iron within hemoglobin in the presence of peroxide. The amount of oxidized TMB (measured at 652 and 450 nm) is proportional to the concentration of Hb. Compared to a previously reported TMB method for measuring Hb, this procedure required: less reagents, shortened incubation time (5 minutes versus 20 min), showed a strong linear correlation (R2~0.99) over a 50-fold Hb concentration range, and was ~100X more sensitive (5 g Hb detection level). However, the assay still had a high inter-assay coefficient of variation (CV), well above the desired <5% value. Furthermore, sample absorbance increased over time until about 30 minutes post-incubation (average 4.68% change in absorbance units), which likely contributed to the high inter-assay CV. The high inter-assay CV and upward drift in absorbance could be caused by either free heme (containing Fe3+) released from Hb, or side reactions with Fe3+ from borosilicate glass tubes. Theoretically, EDTA would improve these values by chelating these iron species. The addition of EDTA decreased average absorbance change over 30 minutes by 92% and decreased average inter-assay CV from 8.06% to 4.03% (n=3). Therefore, this greener/safer TMB-based Hb assay is rapid, sensitive, stable, and exhibits low CV, making it a viable alternative to the toxic Vanzetti's azide-methemoglobin method.

Kathrine Gaikoa, Minsub Chungb , and Moon-Soo Kima
a Department of Chemistry, Western Kentucky University, 1906 College Heights Blvd. Bowling Green, KY 42101
b Department of Chemical Engineering, Hongik University 94 Wausan-ro, Seogang-dong, Mapo-gu, Seoul, South Korea


Efficient detection of specific double-stranded DNA sequences possesses a wide realm of possibilities for point of care (POC) devices. Zinc finger proteins (ZFPs) are one of the most common DNA binding proteins being used to perform base specific double-stranded DNA binding and are extensively studied in biochemistry and biomedicine. However, a new domain known as transcription activator-like effector proteins (TALEs) has recently emerged as an alternative platform DNA-binding.
The TALEs are currently thought to be superior methods for DNA specific binding due to their simplistic structure that increases specificity and has more manipulability which would be advantageous for biomedical applications. They are most often 34 amino acids in length with residues 3-11 and 15-33 comprising of 2 helixes while the 12th residue stabilizes the RVD loop and the 13th specifically recognizes DNA bases. The TALEs were engineered to consist of a capture and detection probe that binds to the stx2 gene that encodes for Shiga toxin in E. coli O157. The detection probe of TALE was fluorescently labeled using Alexa Fluor labeling which allowed the TALE to give an emission when binding of the target DNA occurred. The capture probe was immobilized in a gel solution (AA, ZBA, and VA solution mix) that allowed the capture of the target DNA which was confirmed by the fluorescence of the detection probe. This use of TALEs to detect DNA in a wide range of host organisms has exponential possibilities for detection of pathogens in the presence of large amounts of non-target DNA.

MEAGAN MEDLEY1,2, NICOLE DZUBUK1,3, COLE MALLOY1, EASHWAR SOMASUNDARAM1 and ROBIN L. COOPER1.
1 Department of Biology, University Of Kentucky, Lexington KY
2 Biomedical Sciences, Eastern Kentucky University, Richmond KY
3 Biochemistry, Western Kentucky University, Bowling Green, KY
Acetylcholine (ACh) is an abundant neurotransmitter found in many species across various taxa. In mammals, it is integral in modulating neural circuits underlying processes such as learning, memory, and reward processing. In Drosophila melanogaster, ACh exhibits comparable importance and is known to influence a number of behaviors, including locomotion and feeding; however, the receptor subtypes through which ACh imparts its modulatory influence in this model is poorly understood. The receptors that facilitate synaptic transmission at cholinergic synapses and beyond are divided into two broad subtypes: the ionotropic nicotinic acetylcholine receptors (nAChRs) and the metabotropic muscarinic acetylcholine receptors (mAChRs). Here, we have utilized combined pharmacological and genetic approaches to assess the role of muscarinic receptors (mAChRs), specifically, in regulating two essential behaviors: locomotion and feeding in larval Drosophila. We have revealed that acute and chronic exposure to a mAChR agonist, muscarine, and receptor antagonist scopolamine, significantly alters both behaviors. Moreover, tissue-specific mAChR knockdown also significantly modifies both locomotion and feeding. The results suggest these receptors play an integral role in mediating ACh regulation of these fundamental behaviors. Additionally, we are currently adding to this knowledge by testing the impact on developmental rates with reduced mAChR expression and developmental exposure to receptor agonists and antagonists.
Funded by Kentucky Science and Engineering Foundation (KSEF-3712-RDE-019) at the Kentucky Science and Technology Corporation (RLC); Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under grant number P20GM103436 (MM & ND).

O-linked N-Acetylglucosamine (O-GlcNAc) is a post-translational modification of nuclear and cytoplasmic proteins that is highly conserved in many multicellular eukaryotes. It can modify protein function and it has been shown to be a significant factor in diseases such as heart disease, Alzheimer's disease, as well as play a role in diabetes. O-GlcNAc is regulated by two enzymes; O-GlcNAc-transferase (OGT) which adds the modifications to serine and threonine residues and O-GlcNAcase (OGA) which removes them. The majority of research to date focuses on O-GlcNAc as a cellular stress response and to be a key regulator of early embryonic development. This study focused on using the model organism Drosophilia melanogaster in hopes of finding a scoreable phenotype that could then be used for genetic screens in the future. We hypothesized that O-GlcNAc signaling would play a vital role in the lifespan of Drosophila. We used the UAS-GAL4 system to generate cardiac-specific as well as whole-body OGT/OGA deficient fly lines which were then utilized to determine how altered O-GlcNAc signaling would affect lifespan. After selection against phenotypic markers to ensure accurate crosses, life span was recorded every other day. It was found that removal of OGT from the entire animal (thus reducing O-GlcNAc signaling) resulted in no adult progeny. OGA knockout (thus increasing O-GlcNAc signaling) increased the life span of both males and females.

Blocking muscarinic receptors disrupts memory, via inhibition of adenylate cyclase activity and decreased cAMP concentration in the brain, whereas inhibition of phosphodiesterase (PDE4) increases cAMP concentration. We examined interaction of PDE4-muscarinic receptors on memory during a go/no-go task in rats, using rolipram, a PDE4 inhibitor, and scopolamine, a muscarinic antagonist. Rats were trained on a go/no-go task requiring a lever-press (go) when a cue was presented on the right, but the withholding of a lever-press (no-go) during cue-presentation on the left. Drug effects on correct responses were measured under four conditions: saline, scopolamine, rolipram, and rolipram+scopolamine. Compared to saline controls, scopolamine or rolipram alone increased incorrect responses on both go and no-go trials. Co-administration of rolipram and scopolamine markedly increased incorrect responses, indicating that rolipram failed to reverse scopolamine-induced deficits. Importantly, lever-presses during no-go trials increased following rolipram, scopolamine, or both. Our findings suggest that behavioural deficits reflected impaired memory, not a decrease in activity, and that there was no direct interaction between PDE4 inhibition and muscarinic receptor activation. Given that a go/no-go task depends on the prefrontal cortex, PDE4 inhibition may not directly mediate working memory, but may involve activation of a second messenger system.

Clove oil, used by humans as an essential oil, contains eugenol as an active ingredient. Eugenol acts as a topical anesthetic to remedy pain but the exact mechanisms are still not fully understood. We examined the resulting activity of eugenol on neuronal activity in sensory and motor neurons in crustacean models. Since crayfish, crab and shrimp hearts are neurogenic, we also hypothesized that the heart rate would decrease quickly in these animals. Surprisingly, this study found no change in heart rate despite administering eugenol into the hemolymph to reach 400ppm in crabs or crayfish but some shrimp preparations decreased. The activity of the primary proprioceptive neurons was reduced at 200ppm and ceased at 400ppm for both crayfish and crab preparations when saline containing eugenol was directly applied to exposed sensory organs. Flushing out eugenol resulted in recovery in the majority of the preparations. Administering eugenol to crayfish and crabs both systemically and through exposure in their aquatic environment resulted in the animals becoming lethargic. Direct injection into the hemolymph was quicker to decrease reflexes and sensory perception but heart rate was still maintained. Eugenol at a circulating level of 400ppm decreased electromyogram activity in the claw muscle of crabs. Excessive use of clove oil can result in overdose and cellular toxicity in humans. Overuse of topical eugenol may block proprioception in humans by the same mechanisms as in our model animals. Our next focus is to determine the mechanism of action by intracellular recordings from neurons.

Cisplatin is an anticancer drug which can cause the release of reactive oxygen species (ROS) that kill cancer cells. Curcumin is a compound that can improve the activity of cisplatin against cancer but exhibits poor bioavailability. We decided to investigate whether two synthetic curcumin analogs (curcuminoids), (3E,5E)-3,5-bis[(2-fluorophenyl)methylene]-4-piperidinone (EF-24) and 4-[3,5-Bis[(2-chlorophenyl)methylene]-4-oxo-1-piperidinyl]-4-oxo-2-butenoic acid (CLEFMA), with improved bioavailability, increase the effect of cisplatin against cancer. A cell migration assay was used to test if the curcuminoids enhanced the activity of cisplatin against cancer cell motility using the lung cancer cell line, A549. Cancer cell monolayers were scratched and then incubated in media with control or experimental treatments. The area of the scratch (wound) was measured at two different time points (0 and 24 hours). We also performed an ROS assay to determine if combining curcuminoid and cisplatin treatment affected ROS release. A549 cancer cell samples were treated with controls or experimental compounds and after 24 hours, cell homogenates were collected, treated with a fluorescent ROS indicator and then aliquoted in a 96 well plate for spectrophotometric analysis. Our results suggest that combining either EF24 or CLEFMA with cisplatin reduces cancer cell migration more than cisplatin alone. Also, we found that cisplatin treatment increased ROS release, but the curcuminoids reduced ROS and counteract cisplatin ROS release. Thus, the curcuminoids may promote cisplatin's effect against cancer cell migration but not by modulating an increase in ROS release.

Excessive stimulation of glutamate receptors has been implicated in neural damage and cell death, and it is one of the causes of dementia. Although such over-excitation can be prevented by direct glutamate antagonists, such as memantine, their therapeutic use in moderate-to-severe Alzheimer's disease remains controversial. This study further examined the therapeutic effects of NMDA antagonists on memory, using a simple task. Rats were trained on a fixed ratio 20(FR20), which required 20 lever-presses for each food-pellet reward. MK-801, a NMDA antagonist was tested in conjunction with scopolamine, a muscarinic receptor that is commonlt used in animal models of Alzheimer's disease (AD). Drugs' effects on response latency and task completion were measured under four conditions: saline+saline, saline+MK801, saline+scopolamine, and MK801+scopolamine. Compared to saline-controls, scopolamine impaired behavior by increasing response latency and decreasing lever-presses that earned rewards. MK-801 did not affect performance, but it reversed scopolamine-induced deficits. Our result provide further evidence that in some cases of AD, blocking NMDA-receptors may avert memory impairment by preventing or decreasing cell death due to over-stimulation of NMDA receptors in the brain.

Acetylcholine (ACh) is an abundant neurotransmitter and neuromodulator in many species. In Drosophila melanogaster ACh is the neurotransmitter used in peripheral sensory neurons and is a primary excitatory neurotransmitter and neuromodulator within the central nervous system (CNS). The receptors that facilitate synaptic transmission at cholinergic synapses and beyond are divided into two broad subtypes: the ionotropic nicotinic acetylcholine receptors (nAChRs) and the metabotropic muscarinic acetylcholine receptors (mAChRs). This receptor classification is shared in both mammals and insects; however, both the pharmacological and functional characterization of these receptors within the Drosophila nervous system has lagged behind its mammalian model counterparts. In order to identify the impact of ACh receptor subtypes in regulating the performance of select neural circuits within the larval CNS, we have used a behavioral and electrophysiological approach to assess cholinergic modulation of locomotion, feeding, and sensory-CNS-motor (sensorimotor) circuit activity. We have exposed intact 3rd instar larvae to ACh agonists and antagonists to observe modulation of these behaviors and also expose an intact nervous system directly to solutions containing these compounds to address their influence on sensorimotor circuit efficacy. We reveal that chronic ACh exposure enhances locomotion but reduces feeding behavior and acute application excites a sensorimotor circuit. Nicotine exposure reduces activity through suspected rapid receptor desensitization. Moreover, chronic muscarine exposure reduces locomotion and feeding, but acute exposure enhances sensorimotor circuit activity. These results suggest a role for both nAChRs and mAChRs in modulating these select circuits and illuminates important pharmacological properties of cholinergic receptor subtypes in vivo. Funding Dept. of Biology, UK. Ribble funds (ES), Kentucky Science and
Engineering Foundation (KSEF-3712-RDE-019) at the Kentucky Science and
Technology Corporation (RLC) & personal funds (RLC).

The ketogenic diet is commonly used to control epilepsy, especially in cases when medications cannot. The diet typically consists of high fat, low carb, and adequate proteinwhich producesthe metabolite acetoacetate. Glutamate excitotoxicity is largely implicated in seizure disorders, and therefore is a point of research for control of these disorders. Acetoacetate is heavily implicated as the primary molecule responsible for decreasing glutamate in the synapse; it is believed that the acetoacetate interferes with the transport of glutamate into the synaptic vesicles. The effects of synaptic transmission at glutamatergic synapses was studied in relation to the ketogenic diet in Drosophila larvae for this project. Survival rates, developmental curves, behavioral assays and measure of synaptic transmission were conducted. Higher fat in the diet decreased survival and reduced behavioral responses. We are currently measuring the size of the miniature postsynaptic responses as it relates to a change in quantal response and evoked synaptic responses. We are also addressing the effects on the kinetics of synaptic transmission. We are also assessing the effects of direct application of acetoacetate on synaptic transmission and larval behaviors. This research is significant in addressing the mechanism of action of acetoacetate on synaptic transmission and the developmental effects on neural systems. Funding: Sustaining Excellence-2014 Howard Hughes Medical Institute (Grant #52008116) awarded to the Univ of KY (VM Cassone, PI). The authors confirm that the before mentioned funders had no influence over the study design, content of the poster, or selection of this meeting. Personal funds were also used (RLC).

Brianna K. Ward*, Elizabeth A. Collins, Wesley White, and Ilsun M. White. Neuroscience Program, Department of Psychology, Morehead State University, Morehead, KY 40351.

Nicotine, a psychostimulant, is a direct agonist of nicotinic receptors. Scopolamine, a direct antagonist of muscarinic receptors, impairs memory and is commonly used in animal models of Alzheimer's disease (AD). This study examined the interaction between two cholinergic receptor subtypes in simple memory, focusing on the effects of nicotine on scopolamine-induced behavioral deficits. Rats were trained on a fixed ratio 20 (FR20), which required 20 lever-presses for each food-pellet reward. Once performance reached a behavioral criterion of 60-rewards for 2 consecutive sessions, the drug phase began. Response latency and task completion were measured under four different conditions: saline+saline, saline+nicotine, saline+scopolamine, and nicotine+scopolamine. Compared to saline-controls, scopolamine-alone impaired performance by increasing response latency and decreasing responses that earned rewards, whereas nicotine-alone did not impair performance. Moreover, nicotine partially reversed scopolamine-induced deficits. Our results suggest that nicotine may play a role in improving memory deficits. Given the prevalence of smoking among patients being treated for AD, examining the interaction between the two cholinergic receptors, muscarinic and nicotinic, in memory function is warranted.

Like many other species, fish have been found to represent numerical values in visual discrimination tasks. Little is known, however, about whether fish use number even when other stimulus properties are available. Some researchers propose that animals represent number only as a last resort, while others have found that monkeys will base their decision on numerical value if the numerical ratio is easy to discriminate. In this study, betta fish (betta splendens) were trained to associate food with one of two visual stimuli that differed in both numerical value and cumulative surface area. Non-differentially reinforced probe trials indicated that betta fish were only able to discriminate between stimuli that contained both kinds of information. They performed at chance in discriminating between stimuli that differed only on numerical value, and between stimuli that differed only on cumulative surface area. These results indicate that fish may attend to a combination of these factors during discrimination training, leaving them unable to solve the discrimination with only one of the jointly-learned cues.

Psychology
What influences recognition of emotion among college students?  Shannon Mapes*, Suzanne Savard, Kinetta Crisp, and Ilsun M. White.  Neuroscience Program, Department of Psychology, Morehead State University, Morehead, KY 40351

In social situations, behaving appropriately requires the ability to discriminate emotional expressions accurately.  This study examined factors that influence the discrimination of emotions among college students, using the Diagnostic Analysis of Nonverbal Behavior (DANVA2), which consisted of four subsets of emotion-related stimuli. Each subset included visual stimuli (faces of adults or children) or auditory stimuli (voices of adults or children) conveying four emotions: happy, sad, angry, or fearful.  College students could more readily discriminate adult and child facial expressions conveying positive emotions rather than negative ones. This pattern depended on the type of emotion and the age of the stimulus subject, in that students could discriminate 'sad' faces of children with a high accuracy. No overall sex difference was obtained in discrimination of emotion. African-American students made a greater number of errors in discriminating 'angry' expressions, compared to Caucasians. Given that most stimuli were Caucasians, using emotion of face or voice of out-group members may have influenced discrimination accuracy. Taken together, our findings suggest that among college students the ability to discriminate emotions accurately varies with the nature of the emotional stimuli--stimulus type (adult or child), sensory modality (face or voice), emotional category (positive or negative), and ethnic background. 

Morris1, M., Jones1, V., Prince1, S., Kelling2, N., & Corso1, G.M.
1 Department of Psychology, College of Science, Morehead State University, Morehead, KY
2 Department of Psychology, University of Houston – Clear Lake, Clear Lake, TX
The purpose for this research was to investigate the use video games within the dual-task methodology, and eventually, to extend the use of the games as a diagnostic tool for Attention Deficit Disorder (ADD). The video games used were the worm game and a variant of the Whack-A-Mole (WAM) game. Participants play each of the games separately and the two games simultaneously. For this investigation, the dependent measures are the percent hits and the percent misses for the WAM game when it is a single game and when it is combined with the worm game. Participants (N= 22) participated in two 1-hr sessions. During the first session, participants completed the TOVA or the CPT and then the game. During the second session participants completed the TOVA or the CPT whichever was not competed during the first session and the game. As expected, the number of hits for the WAM single game and the WAM combined game increased from session-1 to session-2 and the number of misses decreased across both sessions. Additionally, the number of hits for the combined game was less than the number of hits for the single game and the number of misses was greater for the combined game relative to the number of misses for the single game. Presently, these results suggest that the video game appears to result in the same type of dual-task performance as traditional laboratory tasks, such as a tracking task combined with a reaction time task.

Research shows that about 75% of us have a bias toward males pursuing careers and females remaining in the home (Project Implicit, 2015). Most people are unaware of this “implicit bias”. However, biases such as these may even influence life and death decisions (Green, et al. 2007). This study aimed to find what effect watching certain gender-related commercials and the experience of working on a simple task with an assigned partner of the same or opposite gender would have on participant's scores on the Harvard Implicit Associations Test. This study involved 34 Berea College students randomly assigned to various conditions. This study found that Berea College students are have implicit biases similar to the general population. This study did not provide support for either of the influences or their interaction on participants' implicit associations. Future research should use fewer independent variables as well as consider ways to increase the strength of the manipulations of the independent variables.

Given that individuals with mental health disorders face barriers in accessing treatment (Mental Health America, 2017), the use of technology to deliver services is increasing. Mental health providers may question the efficacy of this model, given the assumed importance of the therapeutic relationship for comfort in client disclosure (Stiles, 1987). Previous research on clinical populations and the use of technology, such as videoconferencing, found that videoconferencing was just as effective for overall improvement in treating mental health issues and reducing symptoms as traditional face-to-face therapy (De las Cuevas, Arredondo, Cabrera, Sulzenbacher, & Meise, 2006). Outside of clinical populations, results about comfort in self-disclosure in online forums is mixed (Joinson, 2001; Taddei, Contena, & Grana, 2010). This study examined the perceived comfort in self-disclosure in a non-clinical population of 41 college students. Students were placed in either face-to-face (f2f) or video chat conditions where they had to use one of two types of disclosure. Specifically, open frame (impersonal information) or hidden frame (personal information), were elicited in a short conversation with the researcher. Consistent with the hypotheses, two independent main effects were found with increased comfort for both the open frame disclosure and video chat conditions. Although the highest scores were recorded for open frame disclosures with video chat (i.e., an additive effect), the results also indicate that individuals would be more comfortable in disclosing sensitive information via video chat than in the f2f condition. Future research should consider how technology could be used to influence help-seeking and positive therapeutic outcomes in non-clinical, limited-access populations.

Research has demonstrated that mother's ability to think insightfully about themselves and their babies are associated with a wide range of child psychosocial outcomes. The Working Model of the Child Interview (WMCI: Zeanah et al., 1994), used in the current study, has been associated with infant attachment, dyadic interactional quality, and maternal mental health. The WMCI explores the “meaning” a baby has to his or her parent by asking about perceptions of the relationship, the child, and parenting experiences. In this pilot study, the WMCI was administered to three expectant/new mothers' to assess mental representations of their babies. A 7-point reflective functioning scale was used to code the interview and determine a score representing how involved the mother was in thinking about her baby's mental states (i.e. cognitions and emotions). These mothers had followed through on a referral by their nurse midwife for early intervention services. The quality of dyadic synchronous interaction, the mothers' sensitivity to her baby, and the baby's cooperativeness during play was evaluated using the CARE-Index (Crittenden, 1981). Babies ranged between 6 weeks and 4 months. The hypothesis is that mothers with detailed, insightful representations will be more sensitive and will have more harmonious interactions with their infants. These associations will be explored via qualitative analyses. This work was supported by MSU Undergraduate Research Fellowships and an MSU AHRC grant.

Childhood trauma can have a major deleterious impact on individuals, and data is accumulating that suggests this likely creates risk for future offspring. Specifically, parents with higher exposure to childhood trauma have been found to have increased likelihood of frightened, frightening, and dissociated behavior in interactions with their children, as well greater child protection involvement. Increased sensitivity to infant emotional cues, in contrast, may decrease the likelihood of transmitting intergenerational risk. In this pilot study, three expectant/new mothers were given a series of questionnaires, two being specific to trauma: the PTSD Checklist and Adverse Childhood Experiences (ACEs), as well as completed the Adult-Attachment Interview (George et al., unpublished). The AAI is a roughly one-hour assessment of early relationships and experiences with caregivers, as well as their perceived impact on adult functioning and was expanded from the original version with respect to types of exposure and responses to danger (Crittenden and Landini, 2011). These mothers had followed through on a referral by their nurse midwife for early intervention services. The mothers' sensitivity to her baby was evaluated using the CARE-Index (Crittenden, 1981), a three-minute play task between the parent and baby. Babies ranged between 6 weeks and 4 months. The hypothesis is that mothers reporting greater childhood trauma and traumatic symptoms on the questionnaires, and unresolved trauma/loss in the AAI will have lower sensitivity to their infant's emotional cues. These associations will be explored via qualitative analyses. This work was supported by MSU Undergraduate Research Fellowships and an MSU AHRC grant.

This study looked at the effects of top-down and bottom-up processing on memory, as well as the influences of gender and time of day. The thirty-seven participants (22 female & 15 male) were from two Psychology 100 classes, one in the morning and one in the afternoon. Each class was presented with a series of 28 two-digit numbers on powerpoint slides. Each number was followed immediately by an instruction to remember or not remember the number. Half of each set of numbers was presented in silence; the other half of the set with white noise. A significant main effect was found for instruction (F(1, 33)=38.5, p<.001, ɳp 2 = 0.52), which reflects intention's strong influence on memory. A significant main effect for the time of day the class participated in the study (F(1, 33)= 5.44, p<.05, ɳp 2 = 0.13) was also found with students in the morning session performing better than those in the afternoon session. Also a significant three-way interaction between instruction, group, and noise (F(1, 33)= 4.33, p<.05, ɳp2 = 0.11), suggests that the memory processes involved in this seemingly simple task are quite complex.

Most of us hope negative information will not affect how people view us. Affect perseverance theory suggests that negative information has a reduced effect on positive first impressions. On the other hand, affect momentum asserts that people will see us more positively when negative information is presented first. This experiment tests these two theories. If affect perseverance theory is correct, then positive information followed by negative information will result in more positive ratings than in cases where the negative information is presented first. On the other hand, if affect momentum is correct, just the opposite will be found. Participants (N=38) from two General Psychology classes at Berea College were given a packet of four different cases. The participants rated the likelihood of wanting to be friends with each character described in each case on a one to seven Likert scale. The results provided clear evidence for affect momentum by female participants. When given the option to give positive or negative information first, the strategy of giving negative information first could lead to more positive affect when presenting information to females. This strategy could be applicable within job interviews and relationships in general.

The POGIL Approach to Teaching. LUCAS COLLETT* and MEGAN HOFFMAN. Department of Biology, Berea College, Berea, KY 40404.
POGIL, or Process Oriented Guided Inquiry Learning, is a learner-centered method of teaching. POGIL activities strive to imbue students with useful process-based kills, including effective teamwork, communication, and problem-solving. The guided inquiry aspect of POGIL uses a learning cycle that scaffolds knowledge and encourages the students to build their own understanding. In addition to process skills and personally-constructed understanding, another focus of POGIL is to encourage students to use metacognition, i.e. the awareness and understanding of one's thought processes. The POGIL approach is achieved by designing activities based on information-rich models that are explored through carefully-crafted questions. The main goal of the summer project was to create new POGIL activities, while revising others. A series of POGIL activities were developed that focus on aerobic cellular respiration, a pivotal concept in introductory biology. The process of developing and writing the activities will be presented, along with accompanying challenges and future plans.

The POGIL approach, or Process Oriented Guided Inquiry Learning, is a form of active learning that is learner-focused. This approach allows the student to become a more critical thinker and focus on problem-solving as a chance to practice application of what was learned instead of merely obtaining the right answer for a grade. The POGIL approach, which has been used in a variety of STEM disciplines, was implemented in music, with great success. An activity entitled “Biology in Music” was developed to engage students in the anatomy and physiology associated with the voice. The activity follows the POGIL approach by the use of the learning cycle and questions designed to engage students in focused team work and exploration. Exam-based assessments indicated that the students retained the information from the activity and were able to apply it in new situations.

EKU has pioneered the use of content faculty to prepare pre-service K-8 teachers. These courses are dedicated to students in curriculum and instruction with enrollment limited to typically 24-30 students. While courses in biology, physics, and earth sciences are included, this poster describes the chemistry component. We offer two such courses, one for elementary preparation (CHE 104) and one for middle school (CHE 100). We describe here course work consistent with both the three dimensions of NGSS and KY core content for assessment in science, in which a guided-inquiry approach is used, is writing-intensive, and is attentive to both the developmental needs of children and the complex but inadequate science preparation of our students. The following are key benchmarks we have laid in developing groundwork for later work in science: Observation, measurement, use of evidence, conservation of mass, particulate nature of matter, differentiating between mixtures and compounds using simple observation, using inference to understand interactions between materials (electrical and magnetic), clearer ideas about temperature and temperature changes, the kinetic molecular model of matter, the use of simple quantitative approaches to understanding heat flow, and the communication of scientific ideas. This continues to be a work in progress, but we have also notived that even science majors have difficulties with many of these ideas that may be better clarified through these approaches.

The connection between diet and behavior has been well documented in both humans and animal models. The effects of a poor diet can include decreased cognitive ability, aggressive behavior, anxiety, and lethargy in humans. High sugar or carbohydrate diets result in excess secretion of glutamate by the brain, resulting in increased synaptic transmission at nerve terminals. We are developing the Drosophila (fruit fly) model to examine the effects of various diets on behavioral function and development and survival. Diet may also predispose the animals to be able to handle stressors differently. The long term goal is to examine which diet might allow the animal to grow well but also handle starvation and thermal stressors best. Thus, we aimed to test the effect of diet on behavioral response of Drosophila larval behavior. Drosophila are internally analogous to other vertebrates and are similar biochemically and physiologically to humans. This makes them a model system to study the effects of diet on physiological responses related to behaviors, survival and physiological responses. Funding Sustaining Excellence-2014 Howard Hughes Medical Institute (Grant #52008116) awarded to the Univ of KY (VM Cassone, PI). The authors confirm that the before mentioned funders had no influence over the study design, content of the poster, or selection of this meeting. Personal funds were also used (RLC).

Sexual reproduction carries with it many costs in addition to its obvious benefits. Costs associated with mate search, mating displays, and exposure to sexually-transmitted parasites and disease are common in the animal kingdom. Species with particularly aggressive females constitute an additional cost and selection pressure for males advertising to mate. In these animals, males are not only under selection to advertise to the female and outcompete rival males, but they must also correctly gauge female responsiveness to their advertisement. Males of the wolf spider, Tigrosa georgicola, court and slowly approach the large and aggressive females. In this study, we provide a first description of the male courtship display, and then test whether variation in the male display predict mating success by conducting mating trials. In these mating trials, we also quantify the number of female attacks, occurrence of pre-copulatory and post-copulatory cannibalism, and describe female responses to male displays. Our findings help us to understand the evolution of male courtship displays and cognition in species where males are at risk of being cannibalized before having the opportunity to pass on their genes.

The purpose of the bat monitoring program at Clay Hill Memorial Forest (CHMF) is to discriminate between bat species using multivariate analysis. Song Meter SM3 Bat from Wildlife Bioacoustics is the recorder being used within this study. It was hung on a pulley system 65 feet up high in a tree and the calls were recorded on a SD card. These were filtered through Kaleidescope 3.0.1 set at recommendation of USFW Indiana bat monitoring. Analysis was done using SyStat and SigmaPlot to visually plot species separation. The variables used for discrimination include: frequency maximum, minimum, and mean, frequency of the characteristic, frequency of the knee, slope of the characteristic, initial slope, duration of the call, time between calls, time of the characteristic, time of the knee, and the species of each identified call. Gray, Little Brown, Silver-haired, Tri-colored, Big Brown, Eastern Red, Hoary, and Evening are the species that have been positively identified as having been present at CHMF. Small-footed and Indiana had a few identified call series but not enough to show with little doubt that they were in fact present. This scientific study can hopefully assist in formulating a baseline on what bat species are present at CHMF.

A growing body of evidence indicates that increasing levels of human-produced ('anthropogenic') noise in the oceans is having a variety of negative impacts on marine mammals. Establishing the hearing ranges of marine mammals is therefore a crucial part of their monitoring and conservation, and can also help shed light on their ecology, physiology, and evolutionary relationships. Here, we use a combination of morphological data from inner ear labyrinths and existing audiogram data to establish morphological proxies for hearing ranges in globicephaline dolphins. Globicephalinae are a clade nested within Delphinidae that are identifiable by their dark coloration, rounded melon- shaped heads, short snouts, and small numbers of teeth. Audiogram data of hearing frequency ranges for many species within this group are available, making them ideal for producing robust correlations between auditory physiology and morphological measurements. We used a computed tomography (CT) scanner to scan a comprehensive sample covering all globicephaline species. SPIERS and VGStudioMax software were used to extract 3D models of the inner ear labyrinths. Standard measurements and landmarks for ordination were obtained to compare with current audiogram data. Our preliminary analysis shows variation among the inner ear labyrinths. Globicephala melas has a broader cochlear width and lower frequency sound sensitivity compared with Globicephala macrorhynchus, its sister taxon, which has a higher frequency range of hearing sensitivity. Peponocephala electra appears to have higher frequency hearing sensitivity. With further investigation, these data will help us gain a fuller understanding of hearing frequencies among extant odontocetes.

The African lion, Panthera leo, is a vulnerable species and is the only member of Felidae to live in family groups. Therefore, captive-breeding and release programs, such as the one operated by the African Lion and Environmental Research Trust (ALERT), must consider the social cohesiveness among individuals considered for release together. The purpose of this study was to determine to what extent behavioral tendencies are the phenotypic expressions of quantitative traits in P. leo. It is hypothesized that in each behavioral category the percentage of time spent performing activities during daylight hours varies significantly among individuals of the same gender and relatedness value. Behavioral observations were recorded over hour-long focal follow periods every two minutes and subsequently compiled into individual activity budgets. Phenotypes for any given behavioral category are expressed as the percentage of daylight time spent in the activity. A preliminary example of variation between activity budgets can be given by AS5 and his parents, Ashanti and Milo. While AS5 socialized 2% of the time and interacted with the environment 28% of the time, Ashanti socialized 2% and interacted with the environment 33% and Milo socialized 1% and had a 16% interaction with the environment. The relationships between social bond formation and similarity of behavioral phenotypes and relatedness are also being explored. Quantitative genetic analyses will likely be done with the SOLAR-Eclipse statistical program, and social association data will likely be analyzed with the Socprog statistical program.

The effects of many drugs on the phenotype of sleep in vertebrates have not been conclusively proven and are not completely understood. We took 16 female mice and injected (IP) half with 6-OHDA (6-hydroxydopamine) and the remaining half with saline solution as a control. 6-OHDA temporarily desensitizes the sympathetic nervous system (SNS) in the periphery, and the SNS is involved with sleep. We then monitored the effects on their sleep patterns using non-invasive piezoelectric technology. This technology emits a low electric field and senses small movements (including respiration rhythms) from the mice, which can be correlated with sleep/wake patterns. We looked at sleep bout lengths, percentage of sleep in light and dark environments, and sleep histograms to determine if the drug had any effect on sleep. The results showed that there were no outstanding differences between the two experiment groups.

Basement Membranes (BMs) are an evolutionarily conserved specialized form of extra cellular matrix that surround most organs and tissues. Among several other functions, BMs provide structural support to the tissue, and act as selective barriers that prevent tumor metastasis. Previously in our lab several putative BM degraders were identified in a genetic screen. The BM degradation function of SNUTS and CP1 was confirmed by assaying for upregulation of collagenase activity. The SNUTS protein has been suggested to play a role in development of the gonadal stem cell, while CP1 has been found to serve a regulatory function in the development of the Air Sac Primordium (ASP). To further understand the role of these proteins in development we generated antibodies against these proteins. The specificity of these antibodies was confirmed by utilizing western blots and immunohistochemistry on tissues overexpressing and downregulating these genes. Additionally, the antibodies were used to assess the localization of the respective proteins in various tissues. These antibodies represent important reagents that will aid us in our understanding of the function of SNUTS and CP1.

The transcription factor Gli-similar 3 (Glis3) has been shown to play critical roles in the development and maintenance of insulin producing β-cells. Mutations within the human GLIS3 locus are associated with several pathologies including diabetes mellitus. Currently, little is known about the roles Glis3 plays in pancreatic cell-fate specification or its involvement in the postnatal expansion of β-cell mass. Using zebrafish as a model, we have characterized glis3 expression during development using whole mount in situ hybridization and RT-PCR. By 72 hpf, glis3 expression was evident within the pancreas suggesting a possible role in secondary islet generation. These data were supported by morpholino knockdown of glis3, which resulted in decreased insulin transcription at 5 dpf. To better understand the role(s) of glis3 in β-cell maturation and expression, zebrafish were generated that lack functional glis3 expression due to ENU mutagenesis-induced nonsense mutation within exon2 of glis3, resulting in a protein truncated at Thr47 that lacks its DNA binding transactivation domains (glis3sa17645). In order to determine whether glis3 haploinsufficiency resulted in impaired β-cell mass expansion in response to nutrient excess, wildtype and glis3+/sa17645 heterozygotes were fed normal or high-fat/high-glucose diet and were subsequently analyzed for insulin production and blood glucose levels. Collectively, these studies indicate that glis3 has roles in specifying endocrine cell fates during zebrafish pancreas development and may regulate mass expansion in the mature organism. Characterization of glis3 in zebrafish may lead to the identification of therapeutic targets for the treatment of diabetes or other glis3-associated diseases.

Determining the spatio-temporal expression of transcription factor glis3 during development by generating a transgenic zebrafish line using the CRISPR/Cas9 system

Insulin is a hormone produced by the β-cells of the endocrine pancreas that plays an essential role in blood glucose homeostasis by signaling the uptake of glucose by peripheral tissues to be used for energy or stored as fat. Impairment of the insulin response underlies the development of diabetes mellitus. The transcription factor, Gli-similar 3 (Glis3) is important during the development of the endocrine pancreas and plays additional roles as a transcriptional activator of the insulin gene in the mature organ. Multiple pathologies have been linked to mutations within the GLIS3 locus in humans, including type 1 and type 2 diabetes. Although previous research suggests that Glis3 plays a significant role in the specification of β cells during pancreatic development, the spatio-temporal expression patterns of Glis3, its target genes, and its interacting partners remain enigmatic. The zebrafish (Danio rerio) has emerged as a valuable model organism to study development, largely because of its rapid external development and transparent embryos. In order to visualize the glis3 protein during development, a transgenic zebrafish line is being engineered using the CRISPR/Cas9 system to produce fish that express a chimeric protein consisting of glis3 fused in-frame to enhanced green fluorescent protein (EGFP). This line will provide a better understanding of glis3 expression throughout development and can serve as a tool for future experiments to identify glis3 target genes, protein-protein interactions, and characterize molecular mechanisms that could aid in identifying therapeutic targets for the treatment of Glis3-associated diseases.

Cellular enzymes form dozens of post-transcriptional tRNA modifications to increase tRNA stability and function. tRNA modification defects are linked to various diseases, including intellectual disability and cancer. The enzyme responsible for the 3-(3-amino-3-carboxypropyl) uridine (acp3U) tRNA modification, which is found in animals and plants, but not in yeast, is unknown. Because of its high conservation among plants and animals, acp3U is likely to be important for tRNA function. To identify the acp3U enzyme, we are silencing candidate genes in cultured Drosophila melanogaster cells by RNA interference (RNAi), and then analyzing acp3U levels on tRNA from treated cells using primer extension. Candidate genes are identified through BLAST searches of predicted human methyltransferase genes of unknown function that are also found in D. melanogaster and plants, but not in yeast. To date, we have identified 20 candidate genes, treated cells with double stranded RNA (dsRNA) to induce RNAi for over half of these genes, and then analyzed tRNA from treated cells for the presence of acp3U. Because the acp3U modification blocks Watson-Crick base-pairing, its presence causes a primer extension stop which can be detected by polyacrylamide gel electrophoresis. Thus, primer extension of tRNA from cells treated with dsRNA to the gene required for the acp3U modification would result in the absence of the primer extension block seen in wild type cells. Identification of the acp3U gene in animals, including humans, will increase our understanding of the link between tRNA modifications and disease.

Pancreatic beta cell dysfunction in response to chronic hyperglycemia is partially mediated by transcriptional downregulation of Gli-similar 3 (Glis3)

Erin Clayton* and Gary T. ZeRuth, Department of Biological Sciences, Murray State University, Murray, KY 42071


Gli-similar 3 (Glis3) is a Krüppel-like zinc finger protein that plays critical roles in development and in the maintenance of normal physiological functions in a variety of tissues. In humans, GLIS3 deficiency has been linked to a rare syndrome characterized by neonatal diabetes, hypothyroidism, and polycystic kidney disease. In addition, genome-wide association studies (GWAS) have implicated Glis3 as a risk locus for the development of diabetes. While Glis3 is known to play important roles in the specification of endocrine cell fates during pancreatic development, its role in the mature beta cell and the mechanism by which Glis3 dysfunction results in type 2 diabetes remains unclear. We have characterized a rat pancreatic hybridoma cell line (BRIN BD11) that secretes insulin in response to physiologically normal glucose levels but has greatly diminished expression of Glis3. The cell line exhibited significantly decreased levels of known Glis3 target genes, Ins2 and Ccnd2 in addition to the insulin activator, MafA. We investigated the effects of stable Glis3 overexpression on BRIN-BD11 cells and found that Ins2 and Ccnd2 levels were increased in the presence of exogenous Glis3. Interestingly, while chronic exposure of beta cells to high levels of glucose results reduced expression of Ins1/2 and MafA, we found that BRIN BD11 cells were relatively protected from the effects of glucotoxicity and exogenous Glis3 expression partially rescued the phenotype. Collectively, these data suggest that BRIN BD11 cells may be a useful model for the study of beta cell dysfunction preceding the onset of type 2 diabetes.

CELL AND MOLECULAR BIOLOGY

Hepatitis C virus entry inhibitor Aryloxazole modulates Autophagy. JAZMIN M. ESCAMILLA1,2, ZONGYI HU2 and JAKE LIANG 2 Department of Biology, Berea College, Berea, KY 404041 Liver Disease Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institute of Health, Bethesda, MD 208142
Autophagy degrades and recycles cytosolic components and plays a role in the pathogenesis of various diseases including Hepatitis C virus (HCV) infection. HCV can activate the autophagic process which is involved in the replication of HCV. Using high throughput screening, a novel small molecule with aryloxazole moiety (designated as chemotype 6, CT6) was identified as a potent HCV inhibitor. Functional studies demonstrated that CT6 targets HCV at late entry stage (trafficking) of the viral life cycle. We hypothesized that CT6 may interfere with viral trafficking and thus block HCV entry by the modulation of cellular autophagic process. HCV permissive Huh7 cells in culture were used for the study of autophagy modulation. Transfection of autophagy protein plasmids, fluorescent confocal microscopy, and western blot for detecting autophagy marker protein LC3 were used to study the autophagy modulation properties of CT6. Huh7 cells transfected with LC3-RFP and treated with green fluorescent dye bodipy-labelled CT6 showed colocalization of CT6 with LC3. CT6 treatment significantly increased the level of LC3-II in a dose dependent manner. CT6 treatment also appeared to block LC3-II degradation. CT6 appears to modulate autophagy in Huh7 cells through blocking autophagosome fusion with the lysosome. Future studies will characterize the inhibition mechanism of HCV entry by aryloxazole.

Author:
Ahmed Fadil

Coauthor:
Nilesh Sharma

Organization:
Western Kentucky University
ABSTRACT

Rapid applications of titanium dioxide nanoparticles (NPs) in food, cosmetics and medicines have caused serious environmental concerns. There is a lack of understanding of nanomaterial interactions with environment and different biological species. In this background, the investigation was designed to assess the effect of TiO2 NPs exposure on growth, photosynthetic efficiency and oxidative stress in Medicago truncatula. Plants were grown in soil treated with 250, 500, 1000, and 2000 parts per million of NPs for two weeks in a climate-controlled growth chamber. Changes in biomass, element concentrations, and gene expressions related to antioxidant and photosynthetic activities were measured. Results indicate that plant growth was significantly affected with a decrease in their dry weights at all concentrations after 250 ppm. Elemental analysis indicates Ti uptake increased with an increase in concentrations, however, shoot Ti accumulations caused a differential effect in different essential elements (Ca, Mg, K, P, Cu and Fe). While tissue levels of K increased, Ca levels decreased significantly at 250 ppm with insignificant change in Cu, Fe or Mg. Gene expression studies show a significant increase in gene expressions of some of photosynthetic genes at 250 ppm while no significant change or a decline afterwards. Similarly, antioxidative enzyme activities also increased at 250 ppm.

The pathologies associated with neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, and Huntington's disease are due in part to aberrant protein and aggregation and inclusion body formation. Protein aggregates are normally cleared through autophagic mechanisms, and disruption of the constituents of these pathways trigger neurodegeneration in a number of animal models. For example, mutation of the cytosolic puromycin-sensitive aminopeptidase (Psa) in flies results in the age dependent accumulation of protein aggregates and the degeneration of GFP-labeled neurons in vivo.
PAM-1 is the Caenorhabditis elegans orthologue of Psa. PAM-1 has known functions in mediating fertility and lifespan, but little assessment its role in preventing neurodegeneration in worms has been done. We are utilizing a neuron-specific GFP transgene to monitor the development of neurological abnormalities in the GABA neurons in wild-type and mutant pam-1 strains over the course of their lifespan. These abnormalities will be characterized in reference to age of the organisms. We hypothesize that the pam-1 mutant worms will accrue abnormalities at a faster rate than the wild-type worms; and preliminary trials have supported this hypothesis thus far. Abnormalities such as blebs, breaks, and branching are apparent. The development of simple model systems that mimic human pathology may provide useful and powerful reagents in dissecting complex diseases.

Basement membranes are networks within the extracellular matrix (ECM) made up of scaffolding proteins, predominately collagen IV, which provide stabilization and compartmentalization of tissues. Collagen IV emerged roughly 600 million years ago at the divergence of the sponge and Cnidaria most likely resulting from adaptation of the ECM to mechanical stressors, and this protein has been conserved throughout metazoan development strongly suggesting its functional relevance. Unlike many other collagen types, α112 form of collagen IV is widely expressed in many tissues and organs, therefore any disruption of its assembly might systemically perturb ECM structure, thus compromising its integrity risking biological pathologies. Nematostella vectensis is a primitive Cnidarian highly capable of regeneration. Interestingly, it has a basement membrane made of collagen IV which is homologous to the human protein. The aim of this research was to explore Nematostella collagen IV sequence homology with that of the Homo sapiens, and subsequently clone and express it as a recombinant protein in Expi293, a novel suspension cell line characterized by high level of protein expression. This would allow studying of the mechanisms of extracellular assembly of collagen IV and formation of stabilizing crosslinks between NC1 domains. Moreover, this will enable the address of a fundamental question of how the disruption of network assembly affects Nematostella regeneration and normal development.

Many people are suffering from sleep fragmentation (SF) as a secondary symptom of sleep apnea, Alzheimer's disease, Parkinson's disease, and more. Previous research has indicated that one type of glial cell, microglia, are activated in the mouse brain following sleep fragmentation (SF). Another type of glial cell, astrocytes, develop a unique morphology in response to central nervous system (CNS) trauma and increase in numbers in the affected area. To test the effects of acute SF on astrocytes, five male adult C57BL/6j mice were exposed to a SF chamber for 24 hours (60 forced arousals per hour), whereas controls (N = 4) were taken from home cages. Brain tissue was collected from mice following perfusion with ice-cold PBS and then 4% paraformaldehyde. Brain sections were stained for glial fibrillary acidic protein (GFAP) using immunocytochemistry. Astrocytes in the ventromedial hypothalamus and CA3 region of the hippocampus were counted, and processes from five astrocytes in each sample were measured. Acute sleep fragmentation led to astrocyte process elongation in both regions and an increased density of astrocytes in the hypothalamus. These results suggest that acute SF leads to astrogliosis in both the hippocampus and hypothalamus, with greater severity in the hypothalamus.

Cardiovascular diseases (CVD) are the primary cause of global mortality and are disproportionately affecting low and middle income countries (LMICs). The leading risk factor for CVD is hypertension (HTN). While the pathophysiology of HTN is highly heritable, no single genetic determinant is known, and numerous genes appear to mediate this disease. Currently, epigenetic modifications in genes of the renin- angiotensin system (RAS), the hormonal pathway that regulates water and sodium homeostasis, are being identified from a cohort of Kenyans known to have a high prevalence of HTN. Specifically, CpG methylation of AT1 and ACE gene promoters are being analyzed.

Hsp70 is an evolutionarily conserved molecular chaperone responsible for folding proteins, importing proteins into organelles, recovering proteins from aggregations, and assembling multi-protein complexes. Hsp70 overexpression promotes tumor cell proliferation and poor prognosis in cancer patients. Deciphering the regulation of Hsp70 allows us to understand the cell at a basic level and reveal novel anti-cancer therapies.
Autophagy is an important intracellular degradation system whereby cytoplasmic proteins are degraded in the lysosome. There are three independent types of autophagy: macroautophagy, microautophagy and chaperone-mediated autophagy (CMA). Tumor cells utilize autophagy as a means of survival, suggesting that the inhibition of autophagy could be used to develop anti-cancer treatments. While Hsp70 plays a major role in CMA, there has been no established function for Hsp70 in macroautophagy. Recent research from our lab has demonstrated phosphorylation of Hsp70 by the Ulk1 macroautophagy kinase in vitro. Further, by using mass spectrometry we identified ten potential Ulk1 phosphorylation sites on Hsp70. This opens the intriguing possibility that Ulk1 may regulate Hsp70 activity in CMA, connecting macroautophagy and CMA.
In an effort to validate the Ulk1-mediated phosphorylation sites on Hsp70 detected by mass spectrometry, we expressed and purified recombinant Hsp70 peptides containing putative Ulk1 phosphorylation sites fused to GST. We used these fusions as substrates in kinases assay with Ulk1 and assessed Hsp70 peptide phosphorylation by PhosTag gel analysis. We found that Ulk1 phosphorylates Hsp70 peptide with Threonine-111 and Threonine-120. Future studies will use site-directed mutagenesis to determine the role of these phosphorylation sites in vivo.

Polychlorinated biphenyls are persistent organic pollutants widely known to affect the developing brain. Our previous work revealed that allelic differences in the aryl hydrocarbon receptor and cytochrome P450 1A2 (CYP1A2) affect susceptibility to developmental PCB exposure, resulting in cognitive deficits and motor dysfunction. High-affinity AhrbCyp1a2(-/-) mice were most susceptible compared with poor-affinity AhrdCyp1a2(-/-) and wild type AhrbCyp1a2(+/+) mice. Our follow-up studies assessed biochemical, histological and gene expression changes to identify the brain regions and pathways affected. The greatest changes were seen in the cerebellum where a foliation defect was over-represented in Cyp1a2(-/-) mice. In contrast, we found no difference in tyrosine hydroxylase immuno-staining in the striatum. Tyrosine hydroxylase is the rate-limiting enzyme for dopamine production, and this pathway is disrupted in Parkinson's disease. Gene expression patterns varied across the three genotypes, but there was clear evidence of AHR activation. Together, our data suggest that the AHR pathway plays a role in developmental PCB neurotoxicity, but we found little evidence that developmental exposure is a risk factor for Parkinson's disease.

Composite Galactic supernova remnants (SNRs) are a morphological class of sources that exhibit a shell-like radio morphology coupled with a complex X-ray morphology. This X-ray morphology includes a prominent pulsar wind nebula coupled with diffuse thermal emission that is center-filled rather than shell-like, which is typical of Galactic SNRs. As a distinct class of sources, composite SNRs remain poorly studied in comparison to other classes of SNRs and many of their general properties (in particular, their evolution with time) have yet to be studied in systematic detail. We have undertaken an investigation of composite SNRs using archival X-ray observations made by such observatories as Chandra, Suzaku and XMM-Newton: in this talk I present the results of a spatially-resolved spectroscopic analysis of the
composite Galactic SNR 3C 396 (G39.2-0.3) using data from pointed observations made by Chandra and Suzaku. Our results include the detection of an ejecta-rich interior X-ray plasma that features substantial amounts of calcium, a unique discovery amongst Galactic SNRs.

The Cosmic Ray Energetics and Mass (CREAM) instrument is designed to measure the elemental charge of cosmic rays at energies from one to 100 TeV. The instrument consists of a 20 radiation length tungsten calorimeter with scintillating fiber strips (CAL), a silicon charge detector (SCD), and two instruments designed to distinguish cosmic ray electrons from hadrons. The instrument has flown to space seven times from Antarctica using high altitude helium balloons, and was recently modified to fit the form factor required to be attached to the International Space Station (ISS), and was successfully launched and installed in late August, 2017. It is now in flight mode and transmitting data to ground. An overview of the science, instrument, and data status will be given.

The ISS-CREAM instrument was installed on the International Space Station in August 2017. The Silicon Charge Detector (SCD) is a component of the instrument that measures the charge of cosmic rays. A highly detailed model of the SCD was created in the particle physics simulations package GEANT4. A small version of the SCD was created to be tested in the Heavy Ion Beam at CERN in 2016 to determine detector response for ions Helium through Lead. A model of this test SCD was also created in GEANT4 and simulations were run to mirror the conditions of the beam tests. Comparing the simulation data to CERN data provided the information necessary to implement smearing techniques on the full SCD in GEANT4. As a result, simulations for ISS-CREAM can now accurately portray the physical response of the SCD. The detailed process of comparing the data will be discussed.

The Cosmic Ray Energetics And Mass (CREAM) instrument, operating on the International Space Station (ISS) since August 2017, measures the elemental composition of cosmic rays with energies around the so-called “knee” of the spectrum. The calorimeter (CAL), which measures cosmic ray energy utilizing tungsten absorbers and scintillating fibers, was placed in electron and pion beams at the European Laboratory for Particle Physics (CERN). This data is compared to computer simulations of the CAL, first by matching it to electrons, and checking against pion data later. Constants are calculated to understand the amount of additional smearing needed to match simulations to the data, and to determine the calibration constant used to convert from digital units used in the CAL and energy. The simulations will be used to interpret data collected during CREAM's expected three years of operation on the ISS.

DANIELA OLIVERA VELARDE, Berea College,
NOPTREX COLLABORATION — Charge, Parity and Time
reversal (CPT) symmetries are an important aspect of the Standard
Model. One of the outstanding problems in cosmology is the observed
matter/antimatter asymmetry seen in the universe, which requires the
violation of time reversal symmetry (T). The primary goal of the Neutron
Optics Time Reversal Experiment (NOPTREX) is to search for T-violation
in polarized neutron transmission through a polarized nuclear
target. Preliminary measurements were taken on indium and tantalum
resonances at the NOBORU test beam at the Japan Proton Accelerator
Research Complex (J-PARC) to test the functionality of a prototype detector
for the full experiment.

Josephson junctions are natural THz oscillators with great potential for applications as voltage controlled oscillators, electromagnetic signal detectors, or fast switches. Their oscillating frequency depends on their characteristic voltage, which is defined as the product of their critical current (Ic) and normal state resistance (RN). We fabricated high temperature superconducting weak links by overlaying a strip of ferromagnetic material across a narrow bridge made of the superconducting material. We found that the characteristic voltage of the weak link can be as high as tens of millivolts making these devices suitable candidates for fabricating radiation detectors in the THz regime. A summary of our fabrication approach and results will be presented.

While the Kepler Eclipsing Binary Catalog carries over 2800 entries, several are marked as uncertain in nature, or characterized with an 'UNC' flag. We analyze those targets marked as UNC and see if and how many can be modeled successfully as pulsating variables which may be contributing to contamination within the catalog.

Point defects are atomic scale irregularities in crystal structures of solids. These point defects can significantly affect electrical, optical, and mechanical properties of materials. The concentration of a defect depends on temperature, chemical composition, and the difference in enthalpies of the material with and without the defect. Using a thermodynamic model, this study analyzed the temperature and composition dependences of four types of defects in binary intermetallic compounds. It was possible to characterize the enthalpy conditions that lead to temperature- or composition-induced changes in dominant defect concentration. For example, there are ranges of enthalpies for which one type of defect dominates at low temperature and a different type dominates at high temperature for compositions near stoichiometry. The results will allow researchers who calculate defect formation enthalpies using computer simulations to streamline their predictions of temperature-dependent changes in defect-dependent materials properties.

This work is funded in part by the NSF grant DMR 15-08189. We gratefully acknowledge the important contributions of Nicholas Sharp and Yann Couturieux.

In my talk I will present results from Swift and XMM observations of the Narrow Line Seyfert 1 (NLS1) active galactic nuclei RX J0134.2-4258. Swift monitoring was performed between 2007 and 2015 in X-rays as well as in the Ultraviolet.
This source was previously observed by the ROSAT satellite in the 1990s, where it had one of the softest spectrums observed by the satellite. It was believed that the small black hole's high accretion rate may have destroyed its corona through radiation-driven outflows, and that the corona would recover over the following years leading up to the beginning of SWIFT's observation of the source. In addition we will also present the X-ray observations performed by XMM in 2008. Possible models to explain the observations will be discussed.

The “anode heel effect” is a difference in beam intensity at the anode and cathode ends of the x-ray beam due to the geometry of an x-ray tube. In this study we measured the “anode heel effect”. We examined the beam intensity of a Philips DigitalDiagnost X-ray machine in the main radiology department at King's Daughters Medical Center in Ashland, KY. Our results show clearly that the radiation intensity of the beam is greater on the cathode side.

I will report on X-ray, UV, and optical observations of the "Changing Look" AGN IRAS 2322-3843. This galaxy was discovered by Swift in a routine observation in a very low X-ray flux state at the beginning of 2017. We therefore triggered an X-ray observation with XMM and optical spectroscopy with the SALT. The SALT spectrum revealed that IRAS 2322-3843 is one of a rare "changing look" AGN which change their spectral type. While IRAS 2322-3843 was classified as a Narrow-Line Seyfert 1 galaxy based on spectroscopy performed in the 1990s, in our most recent optical spectrum taken by the SALT all broad emission lines have disappeared making it a Seyfert 2 type galaxy. Most-likely what we are seeing in the galaxy is the evolution of a partial covering absorber that not only affects the apparent X-ray spectrum, but also the appearance in the optical.

The APO Galactic Evolution Experiment (APOGEE-1) survey took high-resolution H-Band spectroscopy of 146,000 stars (as of Data Release 12). Of these, 14,840 stars had at least 8 radial velocity (RV) epochs with baselines up to 3 years (~1000 days) making them suitable for orbit fitting. In Troup et al. (2016), 382 of the 14,840 orbits were selected as a gold sample of stars with well-characterized orbits. In this talk, we will discuss our current effort to understand the efficiency of our Keplerian orbit fitting software from the Troup et al paper. In particular, we will analyze a mock catalog of RV curves, and see how well we reproduce their orbital parameters. Knowing the distribution of parameter errors is crucial for using this data set to understand the binary and brown dwarf populations of the Milky Way galaxy. We will also address plans for improving these orbital fits.

The Arduino microprocessor platform has made possible inexpensive prototyping and experimentation. The simplicity of programming the Arduino makes it a great tool for introducing students to computer programming, experimental design, sensor calibration, and data acquisition. We will explain the Arduino platform and show how it can be used in a variety of traditional introductory physics labs as well as some less typical introductory physics experiments.

Large-scale conferences output vast quantities of carbon emissions in a short amount of time, particularly when considering all elements associated with a conference, such as attendee travel, program printing, and venue heating or cooling. The Arctic Science Summit Week (ASSW) was used as a case study to determine whether a carbon calculator and its interpreted results could be used to reduce the carbon footprints of large conferences proactively. Data related to potential carbon emissions for the ASSW were collected or predicated based on previous meetings and known venue operations then input into the Clean Air, Cool Planet Campus Carbon Calculator. Data collected revealed the use of a carbon calculator can provide the information necessary for conference organizers to take steps to reduce a conference's carbon footprint through decisions made during the conference planning process. Specifically, with areas in need of mitigation clearly identified through the use of a carbon calculator, an action plan to reduce a conference's carbon emissions can be developed. Taking measures to reduce the emission of carbon from conferences, offers conference organizers an opportunity to bring attention to the value of assessing carbon footprints, as well as bring awareness to individuals and corporations on the subject of climate change. Additionally, lowering the ASSW carbon footprint is one way planners have committed to help create a positive change for the environment and the people of the Arctic.

Many biomes act as carbon sources and/or sinks. One specific biome of interest in terms of carbon storage, is grasslands. Pasture and rangeland accounts for 27 percent of the total land area in the United States. The soil underneath these grasslands are important carbon sinks. Management of these grasslands affects the carbon balance of the ecosystem. Understanding the movement of carbon in and out of the ground is invaluable management knowledge. This ongoing investigation uses the eddy covariance method to document the carbon and energy balance of a grazed pasture in central Kentucky over a three-year period. Preliminary results have provided insight into the behavior of carbon and energy fluxes through this system as well as driving variables of flux over daily and seasonal cycles.

Despite the many functions and benefits wetlands provide, 80% of Kentucky's historic wetlands have been lost. The Kentucky Vegetation Index of Biotic Integrity (VIBI) was recently developed as a level 3, or intensive, wetland assessment. Although prior validation studies have been successful, more testing is needed before it can be recommended for use across the state. This study collected data from at 48 wetlands from across the state through an intensification of the EPA's National Wetland Condition Assessment during 2016. As part of the intensification, vegetation data were collected and compared with the Kentucky Rapid Wetland Assessment Method (KY-WRAM), which is a rapid assessment method used to measure wetland condition. Despite differences in vegetation sampling methods, results suggest that data collected using the NWCA collection protocols is comparable to data collected using typical VIBI sampling procedures. The 2016 data overall supported the use of the VIBI across most of the state. VIBI scores were significantly higher in Category 3 (or highest condition) wetlands than in Category 2 (moderate condition) wetlands. However, VIBI scores from the 26 sites in the Four Rivers region did not differ between KY-WRAM categories. These results suggest that while the VIBI is well supported in most of the state, more data are needed before the VIBI can be validated and used in the far western part of the state. Low condition sites in particular are lacking from our current dataset. Further validation studies will focus specifically on the Four Rivers region in western Kentucky during 2018.

Over the last decade, sales of organic foods and produce have increased considerably. Though organic foods cost more to produce, consumers believe organically produced foods are safer and pathogen free. Simultaneously, within the last ten years, foodborne disease outbreaks from organic and conventionally grown produce have also increased significantly. Some of the outbreaks may be attributed to use of fertilizers on produce. The objectives of this study are to compare the risk potential of pathogens in commercially available conventional, natural, and organic fertilizers while also evaluating the antibiotic resistance of the acquired pathogens. Fertilizers were sampled categorized as conventional-fertilizers (n=8), natural- fertilizers that are not USDA organically certified but follow organic regulations (n=3), and certified organic- fertilizers (n=7). All samples were analyzed for the presence and enumeration of total Enterobacteriaceae, Coliform, Escherichia coli, and Listeria using Petrifilms and various selective media. Further, the antibiotic resistance of the isolates was assessed using Kirby Bauer Disk Diffusion technique. So far, potential pathogens have been identified in all three categories and the relative risks of potential pathogens per category of fertilizers along with their antibiotic resistance profiles are being analyzed. Being an ongoing study, DNA extraction, PCR Amplification, and gene detection will be the next steps.

With greater awareness of the wide-ranging implications degraded soils have in the food chain, there is growing interest in developing technologies and management practices to improve soil quality. To date, such initiatives are at the forefront of soil science as climate change is expected to alter some of the key soil biophysical and chemical properties. In this study, a two year on-farm demonstration is sought to examine the benefit and advantage of animal manure use as an alternative to chemical fertilizers with substantial potential to supply required crop nutrient needs while improving soil organic content. The on-farm study was conducted at two farms owned by minority farmers at Logan and Madison counties, Kentucky. The broad objective of this study was to determine if using cover crops and/or applying animal manure improves physical, chemical and biological soil properties thereby providing data for economic reasons and incentives for farmers to adopt the practice. Soils at each site was sampled and analyzed for bulk density, organic carbon content, pH, Total P, K, minor elements, soil nitrogen mineralization and soil microbial presence. Comprehensive soil and crop tests and analyses were conducted to identify the key soil quality indicators for monitoring soil health. These measurements were taken prior to, during and after each cropping season. Preliminary findings will be presented.

Adaptive mechanisms to cope with water stress are highly relevant in light of current and predicted climate change. Our work is motivated by an interest in understanding the strategies by which plants endure limited and variable access to water. Vegetative dehydration tolerance (DhT, also dehydration tolerant) is a relatively rare strategy for coping with water shortage that allows photosynthetic tissues to survive considerable cellular drying. DhT provides a unique opportunity to identify key genes, physiological, and ecological traits that can inform management and biotechnological practices aimed at reducing crop and species loss due to drought. Here, we take a within-species approach, minimizing background differences and allowing us to generate a high-quality set of candidate genes that may increase DhT. Our recent studies indicate that Marchantia inflexa, a tropical liverwort, exhibits moderate and variable levels of DhT. Furthermore, M. inflexa can acclimate to different climatic moisture levels, suggesting a plastic component to DhT in this species. Finally, we recently demonstrated that males of this species are significantly less DhT than females. Based on these observations we designed and conducted both genome and transcriptome sequencing studies to characterize sex differences in the presence and coverage of DhT genes, as well as changes in gene expression during dehydration and rehydration across the sexes and tissues. Ultimately, we present a set of candidate genes that may explain underlying differences in DhT.

Policy communication and the Impact of Agricultural Communities on Karst Landscapes: An Example from Phong Nha-Kẻ Bàng National Park, Vietnam

Karst landscapes are vulnerable to agricultural development. Interconnectedness between surface activities and subsurface environments make karst landscapes susceptible to soil erosion and water contamination, both of which increase when agricultural intensification occurs. To mitigate the negative consequences of agriculture on karst landscapes, increased implementation of policy to regulate human activities and communication of these policies is needed. This study occurred in Phong Nha-Kẻ Bàng (PN-KB) National Park, Vietnam, a UNESCO World Heritage site with agricultural communities and mediocre success in protecting its karst terrain. Interviews, observations, and GPS were used to analyze the effectiveness of policy communication and karst protection in the Park. It was found that karst protection policy in PN-KB is minimally communicated and, when communicated, delivered in the wrong way to the wrong individuals. Despite the known harm agriculture causes to karst landscapes, destructive agriculture still occurs frequently and is often supported by government officials. Information on karst landscapes and policies is concentrated among park officials and rarely presented in an informal setting, leaving those in frequent contact with the karst landscape—farmers—without information on the vulnerability of karst terrain to agriculture and the subsequent impacts on human and biological health. In analyzing the situation in PN-KB, general conclusions on policy to protect karst terrain in agricultural regions can be drawn. The successful communication of karst science and implementation of policy to protect the landscape requires information to be presented formally to governing officials and local representatives and then informally communicated social networks of general citizens.

Bird communities are frequently used as bioindicators to assess environmental conditions, including in wetland habitats. We developed an avian index of biological integrity (IBI) for wetlands of Kentucky as an intensive assessment method to supplement an existing rapid assessment method used in regulatory programs. Birds are useful indicators because they are sensitive to environmental changes, abundant in various landscapes, occupy higher trophic levels, and can be sampled in a cost-effective manner. Breeding bird point count data from 140 sites were used to calculate a set of metrics, including avian community measures and guilds based on relative abundance. Metrics were tested for correlation with an independent measure of wetland condition based on landscape and site stressors. High performing, non-repetitive metrics were tested in various combinations to find avian community metrics that best predicts wetland condition. Final metrics were scaled and assembled into an IBI. We found four superior metrics to be correlated with the independent disturbance index. Insectivorous and foliage-gleaning guilds had higher relative abundance at higher condition wetlands, while omnivorous and ground-gleaning guild percentages had higher relative abundance at lower condition wetlands. Guilds represented in these metrics included more species than other metrics tested, creating a greater degree of variation within the guild to correlate to the disturbance index. Previous studies in other regions found similar results with insectivorous and foliage-gleaning guilds being intolerant to human disturbance, whereas omnivorous and ground-gleaning guilds tend to be more tolerant. This cost-effective and time-efficient IBI complements existing assessment tools for wetlands of Kentucky.

Leslie matrices provide a discrete population model that allows for age-group tracking, although it is basically an exponential model. Leslie provided a means of adjusting that model for use on populations with carrying capacities, as long as the same dominant eigenvalue (steady-state age-group distribution) is used throughout the model. The authors have generalized this result so that the initial Leslie matrix dominant eigenvalue can be changed to a different steady-state age-group distribution as the total population nears the carrying capacity. This allows for greater modeling accuracy in our trials.

In this presentation, we will describe how we generated out test data, how we found a nearly-best fit to the data using Leslie's original adjustment, and how we have demonstrated that we can get better results by allowing for a different steady-state age-group distribution than the original Leslie matrix.

The decline of cave-dwelling bats since the introduction of white-nose syndrome (WNS) to North America has led to changes in community interactions as evidenced by spatial and temporal partitioning investigations. Indirect effects, such as disease-mediated competition at the community level, can influence the ability of imperiled species to recover because of competitive exclusion. To further investigate community structure following WNS, we assessed the diet of sympatric species with differential WNS susceptibility using molecular techniques. In western Kentucky, Perimyotis subflavus (Cuvier; susceptible) populations have severely declined following WNS occurrence. Conversely, Nycticeius humeralis (Rafinesque; non-susceptible) populations have increased markedly. We collected guano from N. humeralis (n=38) and P. subflavus (n=9) captured in mist nets during summer 2016. Arthropod DNA was extracted from the guano and a 157 bp target region of insect-COI was amplified. Sequences were analyzed to the lowest taxonomic level provided by the online Barcode of Life Database. Nycticeius humeralis consumed 165 genera belonging to 12 arthropod orders, while P. subflavus ate 92 genera from 8 arthropod orders. All orders consumed by P. subflavus were also eaten by N. humeralis, while 33% percent of all orders occurred exclusively in N. humeralis. Furthermore, N. humeralis consumed 61% of the genera identified in P. subflavus. These data support the potential of increased niche overlap between the two species based on 1) the more generalist habits of N. humeralis and 2) high dietary similarity between N. humeralis and P. subflavus. An increase in niche overlap may suppress the recovery of P. subflavus populations.

Factors Affecting the Diversity of Stream Salamanders in Two Watershed Systems in Central Kentucky
HANNAH ELLIOTT*, MUNTATHAR ALSHIMARY, REBECCA KLEM, JEREMY WILDE, and ROY SCUDDER-DAVIS, Department of Biology, Berea College, Berea, KY. 40404

The Cumberland and Kentucky River watersheds come into close proximity near Berea Kentucky. Both of these watersheds have diverse salamander populations. The purpose of this investigation was to examine stream salamander diversity between the watersheds, and to investigate possible causes for any differences in salamander diversity. Stream salamander diversity did not differ between watersheds, but did differed between habitats with different levels of human disturbance. Streams in non-disturbed habitats in both watersheds had four or five salamander species. Streams in disturbed habitats had only one species.

The differences in salamander diversity between streams in non-disturbed and disturbed habitats does not seem to be related to water quality. An analysis of water quality in terms of chemical pollutants and macroinvertebrate fauna indicated that the water quality in all of the streams was very similar. Differences in water quality indicators were evident in the number and diversity of macroinvertebrate species present, dissolved oxygen content, and conductivity. It is unclear at the moment how these parameters might influence salamander diversity. Further analysis of the data is being conducted to look for patterns in the levels of these parameters in relation to salamander diversity and abundance.

Plants actively defend themselves against insect herbivores, but face a dilemma of costly defenses and unpredictable herbivores. Inducible resistance—whereby defenses are deployed only in the presence of a threat—can help overcome this dilemma. However, plants remain vulnerable during the time necessary to induce defenses. To mitigate this vulnerability, plants may utilize reliable environmental signals as indicators of a future herbivory, allowing them to “prime” themselves. That is, defense priming allows relatively quicker and more aggressive responses once herbivory occurs. While defense priming is well established, fitness costs associated with responding to an herbivore-associated signal are unknown. To address this, we exposed Capsicum annuum and Phaseolus lunatus to a continuous exposure of cis-3 hexenyl acetate (z3HAC), an herbivore-induced plant volatile compound, in a randomized field trial during the summer of 2016. We measured somatic and reproductive output for both species. Relative to their respective controls, z3HAC-exposed C.annuum were shorter, produced fewer flowers and fruits, while z3HAC-exposed P.lunatus had increased floral production, grew taller, and produced more leaves. These results indicate species-specific fitness implications of exposure to a volatile, and therefore management strategies utilizing plant volatiles must account for such species specificity.

Many plants are known to produce and emit distinct bouquets of volatile compounds from their leaves and roots following stress events such as insect herbivory. These volatiles can be detected by natural enemies of herbivores, and by nearby plants. However, little is known about the effect of plant volatiles on seeds. Seeds may lay dormant for years in the soil, and potentially be exposed to a range of plant-derived volatiles. Our objective was to investigate growth and reproductive effects of plants exposed as seeds to plant-derived volatiles. We followed the growth and reproduction in two consecutive generations following a single experimental exposure of plant volatiles to seeds of the model plant Arabidopsis thaliana. Remarkably, both first and second-generation plants were impacted by volatile exposure to the initial seeds. Seed exposure to specific volatiles inhibited primary root growth, lateral root development, yielded smaller plants with lower leaf count and lower fresh biomass of both first and second generations of plants. This work increases our understanding of the long-lasting effect of volatile exposure to seeds on future plants across generations. Moreover, to our knowledge, this is first documented evidence that volatile exposure alone can impact plant growth across generations. Future work will evaluate epigenetic mechanisms for our observed trans-generational effects of plant-derived volatiles on seeds.

Landscape genetics is a rapidly growing field of study that compares patterns of gene flow among populations with habitat heterogeneity across a landscape to infer the interaction between dispersal of individuals and their physical environment. Empirical data generated from a landscape genetics study can be implemented for conservation and management purposes, making the field increasing popular. However, concerns have arisen that the field is expanding faster than the analytic framework that supports it. Multiple methods for generating estimates of the association among habitat types and dispersal (i.e., least-cost paths and resistance surfaces) have been proposed, and there is a debate as to which statistical methods are best for examining the genetic structure on a landscape. We use an integrated empirical- and expert-opinion-based strategy to generate a landscape resistance surface for the California Tiger salamander (CTS), Ambystoma californiense, which is a species of conservation concern. We utilize several alternative analysis methods (e.g., CCA, MRDM, ResistanceGA, and partial Mantel tests) to look for agreement among methods describing the relationship of landscape features and genetic variation. Our analysis revealed variation among methods for describing genetic structure in this CTS metapopulation, but all methods indicated the presence of genetic structure, to some extent, across the landscape. This empirical data set provides both a perspective on habitat management for the CTS and on the suitability of several novel analysis strategies for landscape genetics.

Mountaintop removal mining with valley fills (MTR/VF) often leads to elevated specific conductance within Central Appalachian streams and rivers. Elevated specific conductance is one mechanism hypothesized to be responsible for the decline in stream salamanders in MTR/VF streams, where previous studies have shown reduced abundances and species richness compared to reference streams. Aquatic macro-invertebrates, an important subsidy to stream salamander diet, are also known to decline in streams affected by MTR/VF, potentially limiting the ability of certain salamanders and life stages to persist. In this study, we conducted repeat count surveys of salamanders, examined diets, and collected detail information to water chemistry at 30 sites to answer the following questions: 1) Does stream salamander abundance exhibit threshold values across a specific conductance gradient? 2) What are the effects of specific conductance on salamander diet? We estimated a 56% drop in larval abundance after a specific conductance of 253 μS and a 58% drop in adult abundance after 384 μS. In larval salamanders, the ratio of aquatic to terrestrial prey decreased from 4:1 to 1:1 after 148 μS, the total volume of prey items per stomach decreased by 77% after 113 μS, and the importance values of aquatic prey decreased by 50% after 120 μS. Our results suggest that the reduction in aquatic prey availability is a major driver influencing larval stream salamander occupancy and overall population persistence. Mitigation practices and policies that increase riparian vegetation and canopy cover could provide additional prey subsidies that could restore salamander populations in streams affected by MTR/VF.

Methylmercury is a naturally occurring and toxic form of mercury which may biomagnify through the food chain and bioaccumulate in individuals. This study examines a full body loading and mercury analysis completed for 33 feathers including tail feathers, primary feathers and breast feathers within a single, individual bald eagle (Haliaeetus leucocephalus). Mercury analysis was completed using an AMA 254 Mercury Thermal Analyzer for feather tissue and quill samples. Of all samples analyzed in this project, approximately 46% were quality assurance and quality control samples. Feathers are commonly analyzed for contaminants in avians. Quill analysis has not been reported in any of the related literature. This study is the first study to report analysis of feather quills for mercury in bald eagles or in any other avian species. An implicit assumption of the literature seems to be that the quills are of no significant interest because the mercury levels are low. This study reports elevated levels of mercury in both bald eagle feathers and quills. Feathers ranged in mercury concentration from 7.8 ± 1.6 ppm Hg to 30.4 ± 5.2 ppm Hg. Feather quills ranged from 8.0 ± 2.3 ppm Hg to 34.3 ± 10.9 ppm Hg. The average quill mercury concentration, 15.1 ppm Hg, is actually quite high and is, for individual feathers, correlated with the mercury levels in the vein portion of the feather. The discovery that feather quills contain measurable concentrations of mercury may revolutionize feather analysis and has the potential to expand opportunity for future research.

Since the late 20th century, ecotourism—an alternative to mass tourism with a focus on natural environments—has grown in popularity. Ecotourism destinations are platforms for informal education and exemplify environmental stewardship and conservation. Iceland, an island nation in the North Atlantic, is one area of the world that has seen dramatic growth in its tourism industry over the last several years. Between 2010 and 2015, inbound tourism grew from 488,600 to 1,289,140. Tourists are drawn to Iceland in numbers five times the nation's total population as a result of its unique and diverse landscape, which is characterized by volcanically active terrains, glaciers, waterfalls, coastal communities, and massive fjords. The pressures of economic development have resulted in the continued promotion of Icelandic tourism, and, subsequently, the rapid development of various tourist destinations. This study used a mixed methods approach which included location aware-mobile eye-tracking, post-assessments, and observational analysis to assess visitor experience and behavior in two popular Icelandic tourist destinations: Sólheimajökull and Þingvellir. With these methods, a greater understanding of visitor behavior in these areas was developed. These findings further demonstrate that mobile eye-tracking can be used as an effective tool for data-driven nature-based tourism site development to engage visitors while also promoting education and conservation.

In permanent, multigenerational social groups, certain individuals may have disproportionate influence on group activities and collective decisions. Ties among group members lead to complex and varying patterns of social interactions. We tested whether individuals that have more numerous and stronger relationships with others in the group also have greater influence on group decisions. Specifically, we used Social Network Analysis to explore how social interactions are associated with initiation of group movements and the number of followers in Toque macaques (Macaca sinica sinica). Scan sampling was used to collect behavioral data from a free ranging group of macaques (N= 42) from June–August 2014 in the Mihintale Sanctuary, Sri Lanka. We used USINET 6.528, SPSS 23.0, and SOCPROG 2.5 to perform statistical analyses. The total number of attempts to initiate movement was correlated with the mean number of followers. There was no significant difference in initiation attempts by adult males versus adult females, but the mean number of followers was significantly higher for males than for females. Initiation attempts were significantly associated with both aggression and affiliative behaviors, as well as with dominance scores. However, the mean number of followers was not associated with any of these three traits. Thus, having more interactions is correlated with initiating group movement but not necessarily with success in influencing movement.

Hatchery Creek is a restored stream in Jamestown, KY that drains from Wolf Creek National Fish Hatchery. The previous degraded channel of Hatchery Creek caused large sediment plumes in the Lower Cumberland River and was restored to decrease sediment loss and provide the opportunity for a self-sustaining trout population. We predicted that the increased amount of habitat would increase taxa richness and decrease abundance and biomass. Fish and macroinvertebrates were sampled throughout the year using backpack electrofishing gear for fish, and surber, multi-habitat, and kicknet samples were used to quantify macroinvertebrate density, diversity, and biotic index. Before restoration, taxa richness was 6-8 fish species, dominated by rainbow, brown, and brook trout. The macroinvertebrate community included 8-13 taxa, dominated by low scoring and very-tolerant taxa. Macroinvertebrate density and biomass were over 100,000 ind/m2 and 10 g/m2 respectively in the pre-restored channel. Macroinvertebrate taxa richness increased to 17 taxa, with appearance of new EPT taxa, while fish diversity decreased to 5 taxa. Macroinvertebrate density and biomass in the restored channel have decreased to roughly 1/3th and 1/5th pre-restoration levels respectively. Collector-gatherers remain the dominant functional feeding group in the restored channel, but collector-filterers now make up 33% of overall FFG composition compared to 18% in the unrestored stream. The patterns of fish and macroinvertebrate community recovery suggest new habitat does not result in increased biodiversity within the first year. Further, patterns show that source pools of biodiversity may influence recovery, and that monitoring recovery requires longer time periods.

Understanding fish movement in large systems is important when considering management and control of populations, especially for invasive species. Kentucky Lake is the largest reservoir (~65,00 ha) east of the Mississippi River and stretches 296 km from Kentucky into Tennessee. We used gill nets and electrofishing to collect 96 Silver Carp Hypophthalmichthys molitrix (54 females, mean TL ± SE = 868 ± 7 mm, mean W ± SE = 7,327 ± 311 g; 42 males, mean TL ± SE = 815 ± 9 mm, mean W ± SE = 5,794 ± 250 g), and then implanted ultrasonic tags. We used active, boat-mounted receivers and passive receivers to track these fish throughout the lower 90 km of Kentucky Lake. Tagged Silver Carp also moved into Lake Barkley (an adjacent reservoir) via the canal that connects the two. The maximum distance traveled in a day was 40 km, and mean swimming speed peaked around 4 km/day in late April. No obvious spawning migrations were detected in spring of 2017. All-day tracking suggested that carp activity is crepuscular, with swimming speed peaking near sunrise and sunset. Silver Carp were detected inside the lock chamber at the Kentucky Lake dam, but no fish have left the lake through this route. Fish which were tagged by other research groups have been detected entering Kentucky Lake through the lock. Initial results show that Silver Carp can move long distances over short time periods and movement patterns are variable, even for carp released simultaneously.

Global aquatic ecosystems are negatively affected by numerous mechanisms including habitat destruction, fish overharvesting, climate change, and the introduction of invasive species. Long term monitoring of an ecosystem's fish community, along with abiotic and biotic factors that influence the fish community, is crucial in establishing effective management and conservation strategies. While there are existing fish community monitoring programs across the United States, a long-term monitoring program that includes non-game and non-commercial fish community dynamics has not been established within Kentucky Lake in Western Kentucky. The purpose of this research is three-fold: Establish a power based standardization of sampling effort, to determine which boat electroshocking settings based on three key parameters (catch per unit effort, species diversity, total lengths), and to develop a standardized fish community sampling protocol using the most effective boat electroshocking settings. Four centrally located embayments within Kentucky Lake (Turkey, Anderson, Turner Hollow, and Ledbetter Bays) were sampled with a randomized block experimental design. Within each embayment, 6 separate electrofishing samples were obtained from randomly chosen 500-meter shoreline segments; each sample utilizing 1 of 6 different randomly selected electroshocking wave forms (15, 35, 55, 75, 95, and 115 Hz: 25% duty cycle). Each collected specimen was identified to species and total length of each specimen was obtained. Concurrently with Hancock Biological Station's bimonthly water quality/plankton surveys, this sampling protocol will be used for a long-term fish monitoring program that will track catch per unit effort, body condition, and species diversity of fish communities in Kentucky Lake.

Here we investigate the potential use of solid oil dry absorbent for ignitable liquid sample collection at a fire scene. Extraction of the ignitable liquid sample from a semi-porous surface such as concrete is especially difficult. For this purpose, six different oil dry absorbents were tested. Gas Chromatography coupled with Flame Ionization Detector (GC-FID) was used for identifying the ignitable liquids. Six different oil dries were spiked with different ignitable liquids (charcoal starter, gasoline, kerosene, and diesel). Out of six different oil dries, chromatograms for ignitable liquids obtained from Instazorb and Universal Organic oil dry matched very well with the chromatograms for the respective standards (neat samples). This demonstrates that these two oil dry materials do not interfere with ignitable liquid GC analysis. Currently, analysis of an ignitable liquid sample collected from concrete with the aid of oil dry (Instazorb and Organic) is under investigation.

Secondary organic aerosols (SOAs) form from the atmospheric oxidation of organic compounds that are released into the atmosphere, mainly through photooxidation and OH radical reactions. Reduced sulfur- and nitrogen-containing organic compounds are of special interest due to their abundant release into the atmosphere through agriculture and the burning of fossil fuels and due to their correlation with the formation of SOAs. In this study, kinetic modeling of the atmospheric oxidation mechanisms of dimethyl disulfide (DMDS), dimethyl sulfide (DMS) and trimethylamine (TMA) simulates aerosol chamber experiments to interpret the ionic content of detected particles. The kinetic models were built as systems of differential equations for the atmospheric photooxidation mechanisms of the parent gas compounds. Then, gas phase data from chamber experiments of the corresponding compound in the presence of H2O2 as the oxidant source was simulated to benchmark our models. Experimental data shows a large difference in the production of sulfate ion (SO42-) between DMS and DMDS, as well as a dramatic decrease in SO42- formation when different amines are present, which may indicate rapid aerosol formation or aqueous-phase reactions. Modeling the gas phase data allows accurate prediction of SO2 formation. Observations from modeled data show prompt SO2 formation from DMDS; SO2 production is much slower for DMS. Photolytic dissociation of DMDS appears greatly favored due to its S-S bond, while DMS favors OH addition pathways. Gas-phase data of TMA chamber experiments show a high production of formaldehyde, which suggests that its photooxidation follows a CH3 abstraction.

Recent studies show increasing amounts of harmful organic pollutants in wastewater. Fortunately, organic pollutants can be broken down into harmless end products via photocatalytic degradation. However, more information on the intermediates of these reactions is needed to determine their safety. Ideally, Raman spectroscopy can be used to monitor reactions in real time, 100 times faster compared to the slow process of Gas Chromatography-Mass Spectrometry (GC-MS). However, Raman spectroscopy has low sensitivity, so some molecules that are not strongly Raman active at low concentrations (such as 10-5-M) do not produce high enough Raman intensity in water to be observed. To counteract this dilemma, gold nanoparticles can be used to greatly enhance the Raman intensity of molecules in water, termed Surface Enhanced Raman Spectroscopy (SERS). Gold nanoparticles aggregate to form large clusters of nanoparticles which could precipitate and lower the overall Raman intensity. A capping agent can be used to prevent aggregation by arresting the formation of large clusters. Using the stable (“capped”) nanoparticles, photocatalytic reactions can be monitored by assessing the changes in the Raman spectrum of the molecule being monitored. In this experiment, rhodamine 6G (R6G), a common dye and organic pollutant, and paraquat, a pesticide, were used as the analytes being monitored and sodium dodecyl sulfate (SDS) was the capping agent for the gold nanoparticles. Also, at most acidic pH levels (below pH 5), the capped nanoparticles were unstable yet were stable at basic pH levels (up to pH 12), and a reaction was observed between R6G and OH-.

As the world becomes more industrial, more water is being contaminated. Most pollutants, such as biohazards, inorganic compounds, and sediments, can be eliminated from the water system using widely implemented technology, but organic compounds are complex, making them difficult to remove. Photocatalysts can break down these compounds into less harmful ones, but are not useful on large scale spills because they are dense, causing the particles to sink. Most photocatalysts have a very small particle size, which makes it very difficult to remove from water. To combat these two problems, a photocatalyst was added into an emulsion during microbubble fabrication, which creates a polymer bead as a substrate. Such beads have a large surface-area-to-volume ratio. This gives the photocatalyst maximum amount of access to the sun and the pollutant compared to a sheet. The PDMS beads have a hydrophobic nature and low-density, which makes them float on the surface of the water, allowing for easy removal. The first photocatalyst used in the beads was titanium dioxide (TiO2) because it was inexpensive and has been researched thoroughly. TiO2 beads can removed an organic pollutant from water within a few hours, but it is not very effective in visible light. Zinc oxide (ZnO) was then incorporated into the beads because it has a shown to out preform TiO2.

Photon upconversion is a photophysical process where a material absorbs multiple, low energy photons, and emits a single higher energy photon. Upconverting nanomaterials show promise for applications such as bioimaging, sensing, targeted drug delivery, photodynamic therapy, solar, and security due to their ability to convert lower energy light such as near infrared (NIR) to higher energy, visible or UV. NaYF4 codoped with optically active trivalent lanthanides is commonly used for upconverting applications. In this materiel, a Yb3+ sensitizer is excited with 980 nm photons and the photoexcited energy is transferred to an activator ion (such as Er3+, Tm3+, Ho3+, Dy3+, Nd3+) which emits in the visible or UV. Although traditional methods such as thermal decomposition, solvothermal synthesis, and coprecipitation have been used to produce upconverting nanomaterials with high upconversion quantum yields, these methods have limitations of toxic side products, high reaction temperatures, long reaction times and poor control over the phase and morphology. Liquid-phase laser ablation is a promising alternative for nanomaterial synthesis due to its fast production, use of fewer chemicals, production of fewer byproducts, and control over the product through tuning of the laser parameters. In a typical experiment, a NaYF4:Yb3+/Er3+ target is produced through coprecipitation followed by 532 nm pulsed nanosecond laser irradiation in water. The laser induces formation of plasma plumes which expand, cool, and condense in the water to form particles. The resultant particles show strong red and green emission when excited with 980 nm light. Laser ablation improves upconversion efficiency when compared to the precursor target material. The size of the particles can be controlled by the power of laser used and use of capping agents in the liquid during laser ablation. Other laser parameters such as repetition rate, wavelength, and laser ablation duration can be manipulated to control the resultant nanoparticles.

The objective of our research is to improve to oxidative stability of menhaden fish oil-in-water (O/W) emulsions by incorporating whey protein isolate (WPI) complexes containing xanthan gum (XG) and locust bean gum (LBG) into the aqueous phase. The impacts of sodium chloride (0, 5, 50 mM) and pH (3 and 7) on the stability of these emulsions were investigated on emulsions consisting of WPI, WPI-XG, WPI-LBG, and WPI-XG-LBG. The oxidative stability of these emulsions was tested by measuring lipid hydroperoxide (PV) and thiobarbituric reactive substance (TBARS) concentrations over an 8-week storage period. At 0mM, XG-LBG emulsions displayed the lowest concentrations of PV at both pH values, meaning it had the highest stability. At all salt concentrations, XG-LBG at pH 7 resulted in the highest stability by displaying the lowest TBARS values. XG-LBG did not have higher stability than XG at pH 3, which resulted in the highest TBARS stability at pH 3 at all salt concentrations. Emulsions containing WPI-LBG and just WPI did not show enhanced stability. The improved stability seen in emulsions containing the XG-LBG mixture and just XG at pH 3 can be explained by the electrostatic interaction between the anionic XG and cationic WPI at this pH. Based on the combined PV and TBARS results, it is concluded that WPI-XG-LBG emulsions display the highest oxidative stability of all emulsions, regardless of salt or pH. These conclusions will aid in the distribution of omega-3 polyunsaturated fatty acids into the public diet, allowing for healthier and longer-lasting food products.

Photolysis of nitrate ion in the natural environment produces NO, NO2, and O3, releasing these toxic gases into the atmosphere. Work done by other groups has shown ionic strength dependence of product ratio of aqueous nitrate ion. To better understand the kinetic mechanisms of nitrate photolysis, the effects that ionic strength in solution have on nitrate ion symmetry breaking are needed. Different solvation environments induce nitrate bonding motifs that may be correlated to the product ratio. Fourier-transform infrared spectra of aqueous nitrate–ion solutions were obtained at different temperatures for several total electrolyte concentrations. The electric fields (arising from water molecules and ions in solution) in aqueous potassium nitrate solution distort the trigonal planar shape of the nitrate ion, which may favor a specific initial path of the decomposition of nitrate during photolysis. Van't Hoff plots of the relative peak areas corresponding to the formally-degenerate asymmetric stretching mode reveals the relative energies of the two solvation geometries. The difference in energy between the two geometries is linearly proportional to the ionic strength of the solution. Electronic structure calculations suggest that the more symmetric geometry has an increased stability relative to the less-symmetric geometry in high ionic strength solutions. Thus, the relative amounts of the nitrate ion solvation geometries can be correlated to the amount of products produced during photolysis to help explain the ionic-strength dependence of the product yields. Nitrate geometries at the water—CCl4 interface and aqueous carbonate ion bonding motifs are being investigated to identify pure-water effects.

Pawpaw (Asimina triloba) is a tree fruit native to North America. It is emerging as an alternative high-value niche crop to tobacco among Kentucky farmers. There is literature that claims pawpaw to be a cross-pollinated crop; however, there is evidence of self-compatibility in few varieties. The objective of this study was to determine the self-compatibility and its impact on fruit and seed set in pawpaw. Two advanced selections 'Sunflower', and 'Susquehanna' were selected for this study at Kentucky State University's Research and Demonstration Farm. 250 self and cross pollinations were carried out for each variety with open flowers in April 2016. Flower buds observed after pollination were pinched in May. The fruit clusters along with fruit number were then recorded on three dates, first on 24th May, second on 22nd July and third on 22nd August. The seeds from each treatment were collected, recorded and stored at chilling temperature (4˚C) with wet peat moss for at least 110 days. The stratified seeds, 20 seeds of each cross, were then sown in the greenhouse. The leaf of parent material and newly germinated offspring were collected for DNA extraction. Parentage analysis was done using microsatellite (SSR) DNA markers. The initial fruit set recorded was highest in self-pollination of Susquehanna (65.2%). However, cross-pollination of Sunflower to Susquehanna was significantly higher than the rest in final fruit set (23.2%). The seed set was comparable between self and cross-pollination of Sunflower.

Soil biological monitoring is a potential sensitive indicator of soil ecological stress for early restoration. With increasing cost and shortage of nitrogen fertilizer, there is increased emphasis on the use of municipal sewage sludge (SS) for land farming. However, SS may contain trace-elements that potentially affect soil microbial growth and the enzymes they produce. Soil samples were collected from three Kentucky Counties (Adair, Meade, and Franklin) from areas where SS was applied as a soil amendment for commercial crop production. Hydrolysis of the fluorescein reagent revealed that the total soil enzymatic activities were significantly greater in SS amended soils compared to native soils. The activities of the enzymes hydrolyzing urea (urease; urea amidohydrolase, EC 3.5.1.5), sucrose (invertase; β-D-fructofuranosidase, and p-nitrophenyl phosphate (acid and alkaline phosphatase) were also greater in SS amended soil compared to un-amended native soil. The elevated soil urease and invertase activities (47 and 89%, respectively) as well as acid and alkaline phosphatase activities (23 and 26%, respectively) in soil amended with SS provided evidence of increased soil microbial population and the enzymes they produce.

Atlantic salmon are the most widely produced salmon globally. The Atlantic salmon feed industry still relies on fish meal (FM) as one of the primary dietary protein sources. However, FM has become an expensive and scarce ingredient and its continued utilization in aquafeeds at high levels is economically unfeasible, making the search for alternative protein sources a priority for sustainable salmon production. Although soybean meal (SBM) is the foremost protein feedstuff currently used as surrogates for FM in aquafeeds, Atlantic salmon are highly sensitive to antinutritional factors (ANF) present in conventional soybean meal (CSBM), which limits the inclusion levels of this ingredient in salmon feeds. However, with different processing technologies, including solid-state fermentation and enzymatic treatment, ANF in CSBM can be effectively reduced or completely eliminated improving its nutritional value for monogastric animals. One of these technologies resulted in EnzoMealTM (EM), a product with fewer or undetectable levels of important ANF that has been shown to outperform CSBM in animal feeding trials. Therefore, the objective of this study was to compare the effects of EM inclusion into soybean-based diets for Atlantic salmon (Salmo salar). A 10-week feeding trial was conducted at NOFIMA's land tank facilities in Sunndalsøra, Norway. Four experimental diets were designed to replace FM with three soybean products (SBP): CSBM (Control), soy protein concentrate (SPC), and EM, and one diet containing both SPC and EM. Non-vaccinated Atlantic salmon smolts were stocked into 12 tanks (4 diets, 3 replicates) at 60 fish per tank (10.89 kg/ m3), and fed twice daily to satiation. Preliminary results showed significant differences in growth and hepatosomatic indices of Atlantic salmon in response to the different dietary SBP. Processed SBM significantly increased gut health compared to non-processed SBM, which led to distal intestine soy-induced enteritis. Histology and nutrient digestibility data will also be presented.

Aquaculture is the fastest-growing agribusiness globally and currently supplies over half of all fish and shellfish consumed by humans. In recent years, the aquaculture industry has faced a rapid change in the raw materials used in feed formulations with a departure from marine-based to terrestrial plant-based nutrient sources. Therefore, accurate information on ingredient nutrient composition and availability, and the nutritional requirements of cultured species is a priority for the development of sustainable aquaculture. Most of the nutritional studies on LMB have focused on macronutrient requirements, but amino acids and fatty acids (total n-3) requirements have also been studied. However, the LMB dietary requirement for highly unsaturated fatty acids of the n-3 series (n-3 HUFA), namely eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), remains unclear. Therefore, we conducted a ten-week feeding trial aimed at complementing existing information on LMB responses to supplemental EPA and DHA rich oils. Twelve isonitrogenous (45% crude protein) and isolipidic (12% lipid) diets were formulated to contain different levels (0%, 1%, or 2%) and ratios (20:80 to 80:20%) of EPA and DHA. The diets were fed in triplicate to groups of 15 feed-trained juvenile LMB (13.8 ± 0.2 g initial weight) stocked in 110-L glass aquaria operating as a recirculating aquaculture system. At the end of the feeding trial, the production performance of LMB was unaffected by the various EPA: DHA ratios. However, contrast analyses on performance parameter data reveled that: i) LMB fed diets with either 1 or 2% EPA+DHA displayed higher weight gain and feed efficiency compared to those fed the unsupplemented diets; and ii) LMB fed diets with 1 or 2% EPA+DHA outperformed those fed an EPA/DHA-free and linolenic acid (LNA)-rich diet, suggesting LMB has a limited ability to synthesize EPA and DHA from LNA to meet physiological needs for maximal growth and feed

Applicability of waste-reared Hermetia illucens as an ingredient in the diet of Nile Tilapia, Oreochromis niloticus
Samuel Kessler, Vikas Kumar, Waldemar Rossi
This study sought to define the abilities of the Black Soldier Fly Larvae (Hermetia illucens) (BSFL) as a nutritional aquafeed replacement for Nile Tilapia (Oreochromis niloticus), and to further sustainability by utilizing bioconversion of waste material. An eight-week feeding trial with Nile Tilapia was conducted to determine the nutritional applicability of BSFL reared on dried distiller's grains with solubles (DDGS) in isonitrogenous and isocaloric aquafeed diets, consisting of partial and total replacement for fishmeal (FM), soybean meal (SBM), and oils. Three diets were formulated to contain 32% crude protein (CP) and 8% lipid, with the Control formulated to derive most of its CP and lipid from fishmeal FM, SBM, fish oil (FO) and soybean oil (SBO). The first formulated test diet (BSFLM) replaced all FM and most of the SBM in the Control with BSFL meal. The second (BSFLM+O) was formulated identically to BSFLM except for a complete replacement of FO with BSF oil (BSFLO). Diets were fed to satiation to triplicate groups of 10 juvenile Tilapia in a recirculating aquaculture system (RAS) (stock density 3.08 ± 0.2 kg/m3). Results revealed statistically significant reduction in feed efficiency, protein efficiency ratio, and hepatosomatic index for BSF-based diet groups; though specific growth rate, condition factor, and intestossomatic index of fish were unaffected. No significant dietary effects on health-related blood parameters of the fish were observed. Despite apparently lower nutritional value than FM, SBM, FO and SBO, this study demonstrated that waste reared BSFL-derived feedstuffs are suitable ingredients for the Nile Tilapia diet.

Agricultural practices such as no tillage and plowing influence soil physical and chemical characteristics. In this study, the effects of long-term conventional and no-tillage systems on selected soil properties were determined in a continuous corn system on a Maury silt loam soil. This field for the study is located on the University of Kentucky's Research Farm (Spindletop Farm).  The field was tilled in 1969 from bluegrass sod and the first year's data was in 1970.  Each plot is 20 ft. by 40 ft. and for many years each plot was split with winter cover crop planted to rye or hairy vetch.  The vetch is no long sowed but the rye is continued to be used.  The rye is plowed under every spring in the conventional plots and killed with herbicides in the No-Till plots.  This research field represents 48 years. of continuous no till agriculture under continuous corn.  Each replication contains four rates of nitrogen applied every spring at 0, 84, 168, and 336 kg/ha. In this experiment, the soil samples were collected from the no till and conventional tillage at 0 and 168 kg/ha of N at the depth of 0 to 7.5 cm and 7.5 to 15 cm on June 26, 2017. The samples were analyzed for soil organic carbon (SOC), soil aggregates, C in macroaggregates, soil pH, particulate organic matter (POM), and soil porosity. The results indicate that tillage practice and soil depth were two important factors affecting the dynamics of these soil properties, and conservation tillage practices improve both physical and chemical properties of soil.

Spotted wing Drosophila, Drosophila suzukii, larvae in blackberries bordered by native perennial plants or pasture and treated with biologically based insecticides.

Spotted Wing Drosophila (SWD), Drosophila suzukii, is a new invasive pest of small fruit and fruit tree crops in Kentucky. Female SWDs insert their eggs inside undamaged ripening and ripe blackberries. The larvae hatch, and eat the berry from inside. It has been demonstrated that planting flowering plants and grasses near crops can enhance beneficial insect populations. The objective of this research was to enumerate SWD larvae present in blackberries bordered by native plants or pasture and treated with Grandevo® or Entrust®, two products listed by the Organic Materials Review Institute that can be used in organic systems. This research was conducted at Kentucky State University's Harold R. Benson Research and Demonstration Farm in Franklin County, Kentucky. Blackberry plots were bordered by native perennial plants or pasture. Treatments included Grandevo® foliar spray, soil spray, foliar and soil spray, Entrust® foliar spray and a control using a water foliar spray. Grandevo® and Entrust® were sprayed every other week beginning 7-30 for a total of three times. Grandevo® was rotated with Entrust® while Entrust® was rotated with PyGanic®. Ten berries were collected from each treatment plot weekly and taken to the laboratory. Berries were placed in a sugar solution and the SWD larvae were then counted. Fewer larvae were found in blackberries bordered by native perennial plants than those bordered by pasture overall and later in the growing season when populations of SWD were higher. Results will be discussed relative to native perennial plant bloom periods.

Comparing the abundance of beneficial insects in blackberry plots bordered by native perennial plants and pasture and sprayed with biologically based insecticides.

Conservation biological control is a way to enhance crop production by providing habitat for beneficial insects. It has been found that non-crop plants such as flowering plants and native perennial grasses planted near crops can enhance populations of natural enemies. The spotted wing Drosophila (SWD), Drosophila suzukii, is a new pest of soft-skinned fruit in Kentucky. The female inserts eggs inside ripening and ripe fruit. The larvae hatch and eat the fruit from the inside. The objective of this research was to identify and quantify beneficial insects in blackberry plots bordered by native perennial plants or pasture and treated with Grandevo® or Entrust® which are listed by the Organic Materials Review Institute as treatments for organically grown blackberries. This research was conducted at Kentucky State University's Harold R. Benson Research and Demonstration Farm in Franklin County, Kentucky. Treatments consisted of Grandevo® foliar spray, soil spray, foliar and soil spray, Entrust® foliar spray and a water foliar spray, which was the control. Grandevo® and Entrust® were sprayed 7-30, 8-10 and 8-25. Five sticky traps were placed in each border row and in the middle of each treatment in the blackberry rows. Traps were collected and reset weekly. Traps were brought to the laboratory for identification and quantification. Beneficial insects found included lady beetles, minute pirate bugs, big-eyed bugs, syrphid flies and lacewings. The relative abundance of minute pirate bugs was greater in blackberry plots bordered by native perennial plants than in plots bordered by pasture.

Optimizing Hemp Production through Cultivar Selection and Soil Management Practices
ROSE JOHNSON1*, KIRK W. POMPER1, GEORGE ANTONIOUS1, STEVEN BORST2, JEREMIAH LOWE1, SHERI CRABTREE1, and SHAWN LUCAS1, 1College of Agriculture, Food Science, and Sustainable Systems, Environmental Education and Research Center, Kentucky State University, Frankfort, KY 40601. 2Alltech Crop Science, Catnip Hill Pike, Nicholasville, KY 40356.

Abstract

Industrial hemp is a fiber, oil and seed crop with great potential as a new crop for Kentucky's organic and conventional farmers. A recent Kentucky KRS § 260.850-.869 bill was passed to allow industrial hemp production in Kentucky. The objectives of this hemp research project were to: 1) compare yield of various hemp varieties, 2) investigate the impact of Soil-SetTM and Crop-Set™ on hemp yield, and 3) monitor enzyme activity as affected by Soil-SetTM and Crop-Set™ on field application. There were three trials conducted in 2015 and 2016. Trial 1 was 18 plots (4 by 16 foot each) using a randomized complete block design with four blocks. The treatments were: 1) variety (Finola or Futura 75); 2) control (untreated); 3) Soil-Set® applied at 16 oz per acre at seeding; 4) Grain-Set® applied at 8 oz per acre at vegetative stage 1006-1010 (3rd to 5th leaf pair/5 to 9 leaflets); 5) Soil-Set® and Grain-Set® applied as in treatments 3 and 4, and 6) Soil-Set® applied at 32 oz per acre at seeding. Soil samples were collected from the top 15 cm of each replicate before treatment (six samples total) to establish baseline data. Post-treatment soil samples were collected from every plot every other month between seeding and harvest there was a trend for hemp treated with Soil-Set® and Soil-Set® + Grain-Set® to produce more biomass. Its seems to indicate that, despite weed pressure Soil-Set® + Grain-Set® treatment have increased grain and biomass

We conducted a 10-week feeding trial to examine three soybean meal (SBM) products as surrogates for fish meal (FM) in the diet of largemouth bass (LMB), Micropterus salmoides. Following an incomplete 2 × 3 factorial design including two FM replacement levels (50 and 75%), three SBM types (SBM-A, B and C), and a 35% FM control diet (FM-35), seven isonitrogenous (42 % CP) and isolipidic (12% lipid) experimental diets were formulated. Each experimental diet was fed twice daily to triplicate groups of 15 LMB juveniles (15.3 ± 0.32 g/fish) stocked in 110-L glass aquaria operating as a recirculating aquaculture system. At the end of the feeding trial, survival was above 90% for all treatments and was unaffected (P>0.05) by diet. Feeding rate ranged from 3.4 to 4.1% BW/day and was significantly higher for LMB fed the SBM – C diets. Despite higher feeding rate, fish in the SBM-C treatments displayed significantly lower final weight, weight gain, and feed efficiency compared to those fed SBM-A and SBM-B diets. Both SBM-A and SBM-B could replace 75% of the FM in the FM-35 diet without affecting the production performance of juvenile LMB. We also found significant (P = 0.04) and marginally significant (P = 0.08) detrimental effects on final weight and weight gain of LMB, respectively, when FM replacement increased from 50 to 75%. These effects were likely caused by SBM-C diets alone since no other dietary treatment differed (P>0.05) from FM-35 as analyzed using Dunnett's test. Significant effects of dietary treatments were also observed on some of the analyzed blood parameters of LMB, which will be presented. Dietary FM can be reduced from 35 to 8.8% replaced by SBM-A or SBM-B without detrimental effects on the production performance of LMB. Additional studies are warranted to further optimized SBM-based diets for this species.

This study analyzes differences in antibiotic susceptibilities among isolated bacterial colonies of the same species from a single fish from numerous cases diagnosed at the KSU Fish Disease Diagnostic Laboratory (FDDL). For each case, four identical bacterial colonies isolated from a single fish will be tested for susceptibility to oxytetracycline, florfenicol and sulfadimethoxine + ormetoprim antibiotics. It is hypothesized that consistency in susceptibilities to all three antibiotics will be found among all four isolates from the same fish. This would give credibility to recommendations made by FDDL to use these antibiotics when treating for the infection with medicated feed. If differences occured in antibiotic susceptibilities among bacterial isolates (with the same identification) from the same fish, this would compromise the confidence in FDDL recommendations. In other words, antibiotic susceptibility results would be dependent on the bacterial isolate which happened to be chosen, and results would not have been the same if another isolate had been chosen. Results of this study will either confirm FDDL protocols (especially if consistency in antibiotic susceptibilities is found among bacterial isolates) or show that FDDL protocols may need to be modified to achieve more accurate and consistent results.

Organic farming, farming without chemicals, needs organic fertilizers. Knowledge about recycling waste and adoption of appropriate disposal to enhance and protect soil quality require timely delivery of research and educational technology. An increase of organic waste originated from human activities is a continuous concern. Waste application to soil is proposed as a solution to the disposal problem. Because of the rapid growth in the poultry industry, significant chicken manure (CM) generation will become available in increasing quantities. A field trial area was established in Fayette County for gowing tomatoes using low-cost fertilizers. Tomato (Lycopersicon esculentum var. Mountain spring) seedlings of 52 days old were planted in 30' × 144' beds of freshly tilled soil at eight inch row spacing. The entire study area contained 30 plots (3 replicates × 10 treatments). Each bed was divided into three replicates in a randomized complete block design (RCBD) with the following 10 treatments: 1) no-mulch untreated soil; 2) sewage sludge; 3) horse manure; and 4) chicken manure; and 5) yard waste compost. Each of the 5 treatments was also mixed with biochar to make a total of 10 treatments. Results revealed that yields obtained from CM and CM mixed with biochar were highest, whereas, yield obtained from horse manure was lowest compared to other soil treatments. Total weight of tomato fruits collected after the three harvests revealed CM and CM biochar treatments significantly (P< 0.05) increased tomato yield compared to other treatments indicating a positive effect on the growth and yield of tomato.

Distributing locally grown food fish live may provide aquaculture producers in Kentucky a more profitable market opportunity than selling fish as a commodity. Holding fish live in recirculating systems will be required for this pathway to be successful. In this study investigators evaluated water quality and physiology of Channel Catfish, held live at two stocking densities: 20 kg/ m3 and 40 kg/ m3.
Catfish with a mean weight of 0.76 kg were stocked in recirculating aquaculture systems with a 17 L bead filter after simulated hauling for five hours. Water quality for each holding tank was measured at stocking (Time 0), and daily thereafter (24, 48, 72, 96, 120, 144, 168, and 192 hours). Total ammonia nitrogen, nitrite, nitrate, temperature, dissolved oxygen, pH and conductivity was measured daily while alkalinity, hardness, CO2 and turbidity was measured every other day.
Total ammonia nitrogen increased over time at the higher density. In both treatments, alkalinity and pH declined over time. With this reduction, unionized ammonia peaked at 48 hours and remained below .005 mg/L thereafter. An average water temperature of 25⁰C was conducive to biofilter performance.
With respect to low and high density treatments, mean shrinkage by weight was 8% and 7%; and mortality was 4.6% and 1%. Bites and scrapes were evident on fish during the experiment.

Online shopping is increasingly becoming popular among consumers and businesses (Kotler and Armstrong, 2012 & Rigby, 2012). Regardless of this popularity, levels of awareness for direct-to-consumer market outlets are still unknown. Nor is it known whether online shoppers are willing to learn about such market outlets in their communities. This study uses data collected from a stratified randomly selected sample of 1,205 online shoppers. Results indicate the 38 percent of online shoppers are not aware of marketing strategies for local fresh produce in their area. 43 percent are aware of marketing strategies for organic produce in their area. 20 percent are not aware of any market outlets for local fresh produce in their communities. Age, female, WIC & food stamp, and interests in CSA positively explain the likelihood of learning about direct-to-consumer market outlets. Those who did not attend farmers' markets recently and those who are skeptical to shop fresh produce online are less likely to learn about the market outlets. This study is useful to fresh produce growers and agricultural marketers. It provides an understanding of levels of awareness about fresh produce markets among online shoppers. It also contains critical information that marketers could use to increase the awareness targeting consumers with specific characteristics.

The purpose of this study is to explain interests that online shoppers have in purchasing locally grown fresh produce from traditional offline direct-to-consumer market outlets like farmers' markets, roadside stands, CSA, and agritourism. We used data collected in 2016 from a stratified randomly selected sample of 1,205 online shoppers within the South region of the U.S. who made at least two online purchases six months prior to participating in this study. We found that 20 percent of online shoppers have no interests in attending the offline market outlets. Among the 80 percent who indicated interests, results show that the primary reason they attend or would attend is to purchase fresh fruits and vegetables. The next most important reason is to support local farmers. The third ranked reason is to purchase other items beside fresh produce. Social interactions are ranked the least motivating factor to attend. This study is significant to local farmers, farmers' market managers, and other agribusiness marketers within local food industry. It provides an understanding of interests that online shoppers have in the offline market outlets for locally grown fresh produce. Researchers will find this analysis study useful when furthering knowledge about this relatively new topic.

This study analyzes the effects that the use of best practices for farm management has on farm financial position and profitability. Additionally, it explains the relationships between farm profitability and other characteristics such as size, location, revenue, form (sole proprietorship, partnership, corporation), enterprise (crops, livestock, fresh produce) and owner's characteristics such as age, education, farming experience, and gender. Data for this study was collected through 850 mailed surveys to small and medium-sized farms in Kentucky, as well as electronically, using respondent information obtained through the KDA public directory accessible through their website. Results indicate the use of the best practices for farm management has a positive effect on farm financial position and farm profitability. Furthermore, farm profitability was found to have a positive correlation with farm characteristics such as size, revenue, form, enterprise types, and owner's characteristics. This study is significant to owners of small and medium-sized farms in Kentucky, as well as to Extension agents working together with policymakers.

Investigators evaluated use of salt in holding systems for marketing Largemouth Bass (Micropterus salmoides) as live food fish. There were two salt treatments per experiment: 0 & 4 ppt; and 4 & 8 ppt. Fish were held in individual recirculating aquaculture systems (RAS) without feed for a period of 8 days. Water quality was monitored daily, and blood chemistry was analyzed at 0, 48, 96, 144, and 192 hours after stocking. Blood chemistry was measured with the Abaxis Vetscan VS2 analyzer.

In experiment 1, ammonia, nitrite and nitrate increased over time and pH decreased after Time 0. There was a treatment effect with ammonia, and pH. Blood albumin, globulin, and total protein increased at Time 0, then remained elevated throughout the experiment. Phosphorus and the enzyme ALT decreased over time, while Blood Glucose increased at Time 0, then dropped and remained low for the rest of the experiment. There were significant differences between treatments for blood sodium and plasma cortisol.

In experiment 2, nitrate increased over time, pH decreased after Time 0; and there were significant treatment effects for nitrite, ammonia, and pH. Blood chloride and sodium increased over time, while calcium, ALP, ALT, and potassium decreased. Blood glucose decreased from Time 0 and remained steady. There were treatment effects with blood chloride and CO2.

There was no mortality or apparent weight loss throughout either experiment. In both experiments ALT, glucose, and pH. There were treatment effects for ammonia, pH, and blood sodium.

The number of food borne outbreaks from fresh produce have increased considerably in recent years. Fresh produce is usually consumed raw, without going through processing such as cooking, making it a vulnerable segment that demands microbial safety assessment. Some of the critical environmental factors that contribute to increased risk of contamination by potential bacterial pathogens include the source of irrigation, manure and fertilizers. The latest changes to the Food Safety Modernization Act's (FSMA) Produce Safety Rule (PSR, 2016) require that “all agricultural water must be safe and of adequate sanitary quality for its intended use” (section 112.41) with an E. coli geometric mean less than 126 cfu/100mL, to be tested at least once a year.. In order to offer GAP training and outreach service to small and limited resource minority farmers, microbial quality of agricultural water used on 20 small farms in Kentucky was evaluated. In all, 59 water samples were collected from 2014-2017. Coliform and E. coli count in 100ml of collected water was tested using IDEXX Colilert system. Municipal (n= 45), surface (n=7), and ground (n=7) water showed an average count of 0, 0 and 0.14 cfu/100mL of E. coli. Only one from 7 ground water samples tested positive for E. coli (1 cfu/ml). None of the irrigation water samples tested exceeded the permissible E. coli count. This study will help to develop a research based microbial quality monitoring system for agricultural water and to effectively implement GAPs on small farms in Kentucky and prevent food borne illnesses.