<p> The suicide rate among post-9/11 veterans has become a national crisis. At a staggering number of 22 suicides per day, our wounded warriors are in dire need of psychological assistance. Currently, the treatment interventions used in the rehabilitation of veterans suffering from mental health disorders are limited. Furthermore, these therapy options available to veterans are not realizing adequate improvements in the management of their symptoms. </p><p> This business proposal advocates the utilization of psychiatric service dogs as an alternative intervention to help veterans manage their PTSD symptoms. Rescues for Warriors (RFW), a nonprofit 501(c)(3) organization, aims to pair task trained service dogs with veterans in need of a rehabilitative canine companion. Using evidence-based selection tools, RFW rescues dogs from local animal shelters and uses comprehensive techniques to train them to patient-specific symptoms. This service will be free of charge for all veteran participants, and therefore, marketing and funding will be very important aspects of this business. While this plan does acknowledge the potential challenges in opening charitable organizations, RFW is rigorously committed to increase the quality of life of our nation&rsquo;s heroes and rescue dogs alike.</p>

Crappies spawned in Rough River Lake from April 29 to July 9. Larvae were first collected on May 3 when the lake temperature reached 17 C. A total of 7369 crappie was taken from may 3 to August 1. The maximum number of crappies occurred from May 30 to June 20. The developmental chronology was described for specimens 4.75 mm to 29 mm total length. Meristic characters averaged 10.8 for preanal myomere count, 21.1 for postanal myomere count, 31.9 for total myomere count and 0.62 for preanal length to postanal length ratio. Densities of larvae were low early in the spawn reaching a peak of 147/100 m3 on June 6 and decreased to 10/100 m3 thereafter. Larval stages less than 20 mm were taken primarily at the surface while larger specimens moved to deeper waters. Growth averaged 3.1 mm per week for the 13 week study, 1.43 mm per week for the first 7 weeks and 4.5 mm per week for the last 6 weeks. Major food items of the early postlarvae were rotifers, copepod nauplii and the cladoceran, Diaphanosoma sp. Foods of the late postlarvae included the cladocerans, Diaphanosoma sp. and Simocephalus sp. and larval dipterans of the subfamily Chaoborinae. Primary food items of juveniles were copepods, the cladocerans, Bosima sp., Diaphanosoma sp. and Simocephalus sp. and larval members of the Chaoborinae. No differences in development, meristic characters, density, distribution, growth, food habits and obne and cartilage patterns were observed between white and black crappie 20 mm total length and larger.

Microtubules, isolated from bovine brain, were used to study the effect of ultraviolet radiation on the tubulin-3H-colchicine complex and the polymerizing ability of this complex to intact and disrupted microtubules. Dissociation of this complex was observed upon ultraviolet irradiation and continued to progress as exposure time increased. The binding of this complex to intact microtubules prior to irradiation did not seem to enhance its stability to ultraviolet radiation. There was a significant difference observed at the 0.05 level between the binding of the tubulin-3H-colchicine complex to microtubules in mechanically disrupted and control samples. Statistically, there was no significant difference observed between the binding of tubulin-3H-colchicine complex to microtubules in sonicated and control samples. A slight difference in the electrophoretic mobilities of the alpha and beta tubulins was observed upon electrophoresis of bovine brain microtubule preparations. Various microtubule preparations exhibited different binding levels to 3H-colchicine depending on their degrees of purity. The purer samples exhibited higher binding levels than the less pure samples.

Demands are placed on cattle producers to provide a steady supply of beef at a competitive price. Producers must maximize beef output while minimizing input expenses without compromising product quality. The use of growth implants has become a common practice among cattle producers. The objective of this study was to evaluate the effects of two implant strategies on the average daily gain of suckling calves rotationally grazed on Kentucky – 31 endophyte-infected tall fescue and Kentucky – 31 non-endophyte-infected tall fescue. Eighteen cows with spring calves (N = 18) were used in this study. Calves were grouped by birth date into four blocks. Within each block, calves were stratified by sex and 45d of age body weight into three implant treatment groups for a total of six calves per treatment (n = 6, control 90.3 ± 9.7 kg, zeranol 102.9 ± 10.9 kg, and progesterone (100 mg) and estradiol benzoate (10 mg) 92.4 ± 10.3 kg). Calves were weighed and re-implanted at 129 d of age (84 d after initial implant). Zeranol treated calves were re-implanted using the same implant as the initial implant. Progesterone and estradiol benzoate treated calves were re-implanted after reaching a minimum body weight of 181 kg with either 200 mg progesterone and 20 mg estradiol benzoate or 200 mg testosterone propionate and 20 mg estradiol benzoate depending on sex. Data were analyzed using the REPEATED function in the MIXED procedure of SAS. No interactions were found among sex and treatments for 84d weight gains and 140d weights. There were also no main effects found for 84d weight gains and 140d weight gains. Forage analysis suggested that low crude protein and energy content may have contributed to the low ADG. Low endophyte concentrations may also have played a role.

Ketosis is a metabolic condition that occurs during heat stress, prolonged exercise, fasting, and in obese and diabetic individuals. The major ketone body, β-OH butyrate, affects cellular signaling pathways in a hormone-like manner through its receptor GPR109a. While physiological ketosis is often adaptive, chronic hyperketonemia may contribute to the metabolic dysfunction of diabetes. To understand how β-OH butyrate signaling affects hepatic metabolism we compared the fasting response in control and 3-hydroxy-3-methylglutaryl-CoA Synthase II (HMGCS2) knockdown mice that are unable to elevate β-OH butyrate production, and in GPR109a +/+ and -/- mice. To directly assess the response to ketone signaling we gave intraperitoneal injections of the GPR109a agonists niacin (0.8mmol/kg) and β-OH butyrate (5.7mmol/kg) to HMGCS2 knockdown mice over the final 9 hours of a 16 hour fast. Ketogenic deficient mice showed a more robust increase in hepatic carnitine palmitoyltransferase 1 (CPT1) and uncoupling protein 2 (UCP2) mRNA than control mice in response to fasting. Accordingly, niacin and β-OH butyrate administration decreased fasting CPT1 mRNA expression in knockdown mice, while fasting knockdown UCP2 mRNA and control HMGCS2 mRNA were lowered by β-OH butyrate. GPR109a knockout did not alter the hepatic response to fasting, although the effect of niacin on fasting HMGCS2 mRNA expression was dependent on GPR109a expression. While the role of GPR109a and how chronic hyperketonemia alters β-OH butyrate signaling require further study, these data suggest that endogenous β-OH butyrate signaling during a fast regulates the transcript levels of hepatic genes directly involved in its own synthesis.

Mastitis represents a major issue within the dairy industry responsible for economic loss via decreased animal productivity and associated veterinary costs. Currently, there is a push to identify a phenotypic innate immune response that will yield dairy cows with an enhanced resistance to mastitis. Bovine dermal fibroblasts were used as a cell model to measure the response of individuals to Gram-negative bacterial stimuli through the TLR4 signaling pathway. Fibroblast cultures were isolated from 15 dairy heifers at 5, 11, and 16 months of age in order to determine the variability in responsiveness to LPS as well as to monitor the development of the innate immune response in calves. These individuals displayed a large range in response to LPS as measured by IL-8 production. In addition, response within individuals increased dramatically with age. To determine the cause behind this, DNA methylation was investigated as a potential player in the variation in response described both within an individual over time as well as across individuals. Fibroblast exposure to 5-aza-2'-deoxycytidine, a DNA demethylating agent, increased the cellular response to LPS, but more so in cultures that had previously displayed low responding phenotypes. This suggested that DNA methylation acted as an inhibitor of the innate immune response, and may be responsible for some degree of the variation seen in the LPS response. To determine the effect of epigenetic factors on this response, microarray analysis was conducted on RNA isolated from cells either having been epigenetically modified (DNA demethylation and histone hyperacetylation) or without undergoing any epigenetic treatment. This analysis identified 1,758 genes with altered expression due to epigenetic modification. To focus on DNA methylation's role, methylated CpG island recovery assay (MIRA-Seq) libraries were created from fibroblasts to investigate differential methylation from a group of the same individuals sampled at 5 and 16 months of age. In addition, transcriptomic data were generated by RNA-Seq from fibroblasts collected from the young and older samples and exposed to LPS for 0, 2, and 8 hours to characterize age-associated changes in the innate immune response. Cultures from older animals were much more responsive to LPS as indicated by greater expression of IL-8, IL-6, TNF-α, and CCL20 at various times in response to LPS. TLR4 and CD14 were more highly expressed in older cultures, suggesting these fibroblasts are more able to detect the presence of LPS. Analysis of the bovine fibroblast methylome revealed methylation with remarkable stability except for 20 regions along the genome undergoing major shifts due to age. Similar data were collected from fibroblasts isolated from different individuals displaying either a low or high responding phenotype resulting in 843 regions with differential methylation between groups. This suggests that DNA methylation may be playing a role in both the age-dependent and between animal differential responses to LPS, and also gives the first in depth look at the bovine fibroblast methylome and its stability over time.

The goal of the work presented herein was to further our understanding of Bovine Leukocyte Antigen (BoLA) class I diversity of Holstein cattle and develop tools to measure class I restricted T cell responses to intracellular pathogens such as foot and mouth disease virus (FMDV) following vaccination. BoLA is a highly polymorphic gene region that allows the bovine immune system to differentiate pathogen-infected cells from healthy cells. Immune surveillance by CD8+ T cells plays an important role in clearing viral infections. These CD8+ T cells recognize BoLA class I molecules bearing epitopes (antigenic peptides) of intracellular origin in their peptide binding groove. Polymorphisms in the peptide binding region of class I molecules determine affinity of peptide binding and stability during antigen presentation. Different antigen peptide motifs are associated with specific genetic sequences of class I molecules. In order to better understand the adaptive immune response mediated by BoLA molecules, technologies from human medicine such as high-throughput sequencing, biochemical affinity and stability assays, tetramers and IFN-γ ELIspot assays could be applied. Therefore, it was hypothesized that we can translate these technologies from the study of human T cell responses to the study of cattle immunity. The first objective was to establish a comprehensive method for genotyping BoLA of Holstein cattle by using Illumina MiSeq, Sanger sequencing and polymerase chain reaction sequence-specific primers (PCR-SSP) (See Chapter 2). This is an important first step in order to study the BoLA restricted immune responses following FMDV vaccination. The second objective was to define the FMDV capsid protein peptide repertoire bound by BoLA class I molecules using bioinformatics and biochemical affinity and stability assays to facilitate the identification of T cell epitopes (See Chapter 3). The third objective was to demonstrate clonal T cell expansion for specific epitope polypeptides using ex-vivo multi-color flow cytometric MHC-epitope complexes (tetramers), followed by IFN-γ production measured by an ELIspot assay to quantify and define the antigen specific response of Holstein cattle to FMDV vaccination (see Chapter 4). In this, my dissertation studies aimed to improve our understanding of the BoLA class I restricted T-cell responses to candidate FMDV vaccines in Holstein cattle. In this manner, my research will improve animal health through the production of assays for characterizing the bovine immune response to intracellular pathogens and enhance vaccine design leading to improved biologicals to protect cattle from devastating infectious diseases.

Master of Science / Department of Animal Sciences and Industry / J. Ernest Minton / Mitchell Ricketts / Cattle and other livestock have been identified as leading sources of injuries to workers in agriculture. Cattle handling injuries can be serious and often appear to be under-reported [superscript]3,[superscript]4. Many of these injuries involve predictable patterns of interactions among victims, animals, and fixed farmstead structures or gates. There has been some progress toward developing safer facility designs and work procedures, but continuing reports of injuries suggest further efforts are still needed. The present study focused on worker injuries that involved the interaction of three elements: (a) cattle, (b) cattle handlers, and (c) farm structures or equipment—including swinging gates and stationary barriers. The goal of the study was to identify opportunities for injury prevention. The source of injury cases was the Consumer Product Safety Commission’s National Electronic Injury Surveillance System (NEISS)[superscript]29,[superscript]45. We believe this is the first report of cattle related injuries based on NEISS data. We selected the NEISS database for this investigation because it includes product codes for many farmstead barriers such as fences, walls, and doors. The database was also selected because it contains brief narratives that help to describe the circumstances of each incident. Predictable interactions between humans, animals, and farm structures led to many of the cattle handling injuries reported in the NEISS database. In almost 30% of cases, cattle pushed workers into structures such as fences, gates, posts, and walls. In another 16-19% of injuries, cattle struck gates and other objects, propelling them at the victims. These percentages are similar to findings reported in previous studies that drew on data from New York hospitals[superscript]10, news reports in the central United States[superscript]5, and workers compensation cases in Colorado[superscript]3,[superscript]4. In all, gates and other physical barriers contributed to about 45% of cattle handling injuries in the present study.

Master of Science / Department of Animal Sciences & Industry / Elizabeth A. Boyle / Quality attributes of vacuum packaged, skinless smoked sausage made with a combination of pork, turkey, and beef, cooked to 64, 68, or 72°C internal endpoint temperature following USDA FSIS Appendix A, and displayed at 4°C for up to 120 days under light emitting diode (LED) and fluorescent (FLS) lighting were evaluated. External color, pH, thiobarbituric acid reactive substances (TBARS), proximate analysis, reheat yield, and sensory attributes were measured on day 0, 90, and 120 of display. Purge amount and color were measured on day 10, 90, and 120. Product was displayed in LED or FLS retail display cases set to the same operational and temperature profiles. Lighting type had no effect (P>0.05) on any of the measured attributes. Instrumental external color was less (P<0.05) red by 0.63 units in product thermally processed to 64°C than product processed to 68°C. Product cooked to 72°C was less (P<0.05) yellow externally compared to those processed to 64 and 68°C. Purge color lightness increased (P<0.05) in product thermally processed to 72 compared to 64°C. Purge was more red by 0.36 units (P<0.05) on day 120 compared to day 10. Yellowness of purge color increased at 72°C compared to 64°C by 0.66 units. Purge was more yellow (P<0.05) on d 120 compared to d 10 and 90. TBARS values decreased (P<0.05) from 0.70 mg of malonaldehyde/100g on day 0 to 0.35 and 0.23 on d 90 and 120, respectively. Sensory panel scores showed that flavor intensity decreased (P<0.05) as day of storage increased, and saltiness decreased from d 0 to d 90. Purge content increased (P<0.05) from 1.45% to 1.90% in products cooked to 64 and 68°C, respectively. The amount of purge increased (P<0.05) from 1.58% to 1.92% on day 10 and 90, respectively. While there were slight changes found in quality characteristics of smoked sausages during storage, many of these were minimal. Processors could reduce their internal endpoint temperature following USDA FSIS Appendix A guidelines with minimal effect on product quality. Vacuum packaged pre-cooked smoked sausages could be displayed under LED or FLS lighting with no effect on product quality.

Master of Science / Department of Animal Science and Industry / Barry Bradford / During the past decades, the dairy industry has been challenged to find alternative approaches in order to feed cows without affecting their performance or increasing production costs. To accomplish these objectives, some options that have been implemented are the inclusion of short chain carbohydrates to replace starch and the addition of synthetic supplements to increase feed efficiency. In order to assess the impact of these strategies, an experiment was conducted to evaluate productive responses of lactating dairy cattle when they received sucrose and/or exogenous amylase in low starch diets. The results indicated that milk production, milk component profile, and feed efficiency were not significantly altered by the use of the enzyme, sucrose inclusion, or the combination of both. Comparing these results with the literature revealed apparent inconsistencies in responses to the inclusion of sugar in dairy rations. For that reason, a meta-analysis was performed to determine the impact of different sugar sources on milk production, and also to evaluate the impact of other dietary factors on response to dietary sugar. The results indicated that dry matter intake responses were significantly (P < 0.05) affected by an interaction between added sugar and dietary forage neutral detergent fiber content, but overall, dry matter intake tended to increase when sugar replaced corn grain in diets. Energy corrected milk was not affected by dietary sugar, but milk production showed a tendency to respond to treatment, dependent on an interaction between added sugar and rumen undegradable protein. In summary, sugar inclusion may promote small increases in dry matter intake, but the impact on milk production is inconsistent; both factors may be influenced by the diet to which sugar is added.