Doctor of Philosophy / Department of Diagnostic Medicine/Pathobiology / Wenjun Ma / Influenza A virus (IAV) is an enveloped, segmented, negative-sense RNA virus that infects avian species and mammals. Its segmented feature enables antigenic shift which can generate novel IAVs that pose a threat to animal and public health due to lack of immunity to these viruses. Pigs have been considered the “mixing vessels” of influenza A viruses to generate novel reassortant viruses that may threaten animal and public health. Therefore, it is necessary to understand the pathogenicity and transmissibility of newly emerged reassortant viruses in swine. Adding to this complexity is the newly identified bat influenza A-like viruses which have roused interest in understanding the evolutionary history and pandemic potential of bat influenza. At least 10 different genotypes of novel reassortant H3N2 IAVs with gene(s) from 2009 pandemic H1N1 [A(H1N1)pdm09] have been identified in pigs in the United States. To date, only three genotypes of these viruses have been evaluated in animal models leaving the pathogenicity and transmissibility of the other seven genotype viruses unknown. We showed that reassortant viruses with genes from A(H1N1)pdm09 are pathogenic and transmissible in pigs. Further studies showed that avian-like glycine at position 228 of the HA receptor binding site is responsible for inefficient transmission of the reassortant H3N2 IAV with five A(H1N1)pdm09 genes. Studying the recently discovered IAV-like sequences from bats has been hindered by the lack of live virus isolation or culturing. Using synthetic genomics, we successfully rescued modified bat influenza viruses that had the HA and NA coding regions replaced with two classical IAVs. Additional studies were performed with truncations on NS1 protein and substitution of a putative virulence mutation in bat influenza PB2. Virus reassortment experiments demonstrated that bat influenza has limited genetic and protein compatibility with other influenza viruses; however, it readily reassorts with another divergent bat influenza virus. Taken together, our results provide insights into the pathogenicity and transmissibility of novel reassortant H3N2 IAVs in pigs. It also indicates that the bat influenza viruses recently identified are viable viruses that pose little pandemic threat to humans. Moreover, they provide new insights into the evolution and basic biology of influenza viruses.

Master of Science / Department of Diagnostic Medicine/Pathobiology / Richard Hesse / Bovine Coronavirus (BCoV) is known to cause enteric and respiratory diseases, such as calf diarrhea, winter dysentery, calf respiratory disease, and bovine respiratory disease complex (BRD). All of these diseases are believed to be caused by the same genotype of BCoV. BCoV exhibits tissue tropism for both the gastrointestinal and respiratory tracts. This tropism is due to 9-O-acetylated sialic acid receptor on both epithelial cells in the respiratory and enteric tract. Currently, the only vaccine available for BCoV targets the enteric form of the disease. This study addresses the hypothesis that antibodies from the enteric form of the disease can cross neutralize the respiratory form of the virus. Data from surveillance studies suggest that BCoV is one of the major contributors to BRD, for which there is no currently approved vaccine for the respiratory form of the disease. Our approach to answering this question is to sequence and analyze the complete genome of 11 respiratory and enteric coronavirus isolates using next generation sequencing (NGS). Following the NGS, viruses were selected based on phylogenetic analysis and ability to grow and be maintained in cell culture. These viruses were then be used as serum neutralization indicator viruses in SN assays. 147 bovine serums submitted to KSVDL were used to determine if there are any serological differences between the immune response to respiratory versus enteric viruses based on the antibodies produced by the animal. The overall results show that there are few differences between the enteric and respiratory isolates at the genomic level and the serological response from the animal to these viruses. The differences between enteric and respiratory virus will need to be further addressed and analyzed to conclude if there is a noteworthy difference between the viruses with different tropisms. Other factors, such as host immune response and environment, are believed to be involved in the virus tropism to certain areas of the body.

Master of Science / Department of Clinical Sciences / Gregory F. Grauer / Prostaglandins play many important roles in the kidney including regulation of renal blood flow, glomerular filtration, renin release, and sodium excretion. Upon activation of the renin angiotensin aldosterone system (RAAS), prostaglandin upregulation becomes critical to offset the vasoconstrictive effects of norephinephrine, angiotensin II, and vasopressin. Nonsteroidal anti-inflammatory drugs (NSAIDs) produce both their beneficial and detrimental effects through inhibition of the cyclooxygenase enzyme and subsequent interference with prostaglandin production. Healthy canine kidneys express both COX-1 and COX-2, although basal COX-2 expression in dogs is significantly higher than in other species. Nonsteroidal anti-inflammatory drugs that spare COX-1 have exhibited less gastrointestinal toxicity, but no NSAID has been proven safe for the kidney. The kidney is the organ with the second highest reports of adverse drug events, which is usually manifested as functional changes. However, structural changes including renal papillary necrosis, can occasionally be observed. Dogs with chronic kidney disease could be expected to be at increased risk for NSAID-related adverse drug effects. As nephrons and renal reserve are lost in chronic kidney disease, the canine kidney becomes more dependent on COX-2 for production of prostaglandins. Inasmuch as the prevalence of both CKD and OA increases with age, it is expected that many dogs being treated with NSAIDs for OA will have loss of renal reserve and/or early stage CKD. If administration of an NSAID is required for long term treatment of osteoarthritis, frequent monitoring of blood pressure and renal parameters, as well as hepatic enzymes are recommended.

Doctor of Philosophy / Diagnostic Medicine/Pathobiology / Steven S. Dritz / The overall goal of this dissertation was to evaluate the effects of different amino acid (AA) levels on performance of pigs under commercial conditions. To reach this objective, a total of 12 experiments were conducted. For the sow research, 1,102 highly prolific sows were used to determine the effects of AA and energy intake during late gestation on piglet birth weight and reproductive performance of sows. Weight gain depended on the energy and AA intake levels while sows fed increased amount of energy had increased stillborn rate; however, there was no statistical differences due to energy intake in stillborn rate of gilts. The modest increase in individual piglet birth weight is due to energy rather than AA intake during late gestation. Pre-weaning mortality was reduced in piglets suckling from sows fed high AA diets during late gestation while subsequent reproductive performance was not affected. With recent advances in statistical computing capability, linear and non-linear mixed models were refined to estimate the AA ratio dose-response relationships. Then, 4 experiments using 2,420 wean-to-finish pigs were conducted to validate the methods for estimating the standardized ileal digestible (SID) AA to lysine (AA:Lys) ratio requirement. Subsequently, 7 experiments using 7,562 pigs were conducted to estimate the SID tryptophan (Trp) to Lys and Valine (Val) to Lys ratio requirements of wean-to-finish pigs. In 11- to 20-kg pigs, optimum SID Trp:Lys ranged from 16.6% for maximum mean G:F to 21.2% for ADG. In 30- to 125-kg pigs, optimum SID Trp:Lys ratio ranged from 16.9% for maximum mean G:F to 23.5% for ADG. However, 18% SID Trp:Lys captured 96 and 100% of the maximum mean ADG and G:F for finishing pigs, respectively. In 25- to 45-kg pigs, optimum SID Val:Lys ratio ranged from 72.3% for maximum mean G:F to 74.4% for ADG with 99% of the maximum mean ADG and G:F at approximately 69% and 65% SID Val:Lys ratio, respectively. In conclusion, optimum SID Trp:Lys and Val:Lys were consistently higher for ADG than G:F. This finding is critical for conducting economic evaluations and reference tables such as NRC (2012) should consider presenting requirement values for different response criteria.

Doctor of Philosophy / Diagnostic Medicine/Pathobiology / Michael D. Apley / Bovine respiratory disease (BRD) remains a major disease in beef production systems. The administration of antimicrobials for both the control and treatment of acute BRD is common. According to most published accounts, antimicrobial resistance among BRD pathogens is increasing; therefore, judicious antimicrobial usage is vital for continued efficacy. The introduction of a novel antimicrobial class has not occurred for well over a decade, therefore it is paramount to maximize efficacy of the antimicrobials currently available. The challenge is targeting the perfect scenario: maximizing clinical efficacy while minimizing antimicrobial resistance. The host-pathogen-drug interaction is very complex and despite current sophisticated technology, this interaction is still not well understood for many infectious diseases. This dissertation work sought to investigate the effects of the administration of a macrolide for both control and treatment of acute BRD on the prevalence of resistance among isolated Mannheimia haemolytica, Pasteurella multocida, and Histophilus somni. Whole genome sequencing of M. haemolytica was utilized to investigate the presence/absence of macrolide resistance genes and their relationship to the observed minimum inhibitory concentration. Cattle were sampled (plasma and pulmonary epithelial lining fluid) after administration of gamithromycin for drug concentration analysis. A non-linear mixed effects approach was used to fit a compartmental model to the resulting sparse pharmacokinetic data so that a complete time concentration curve could be simulated. From these curves, the CMAX and AUC were measured and used to calculate standard PKPD indices using the MIC values of the isolated bacteria. Clear associations between the use of gamithromycin for control and treatment of BRD and a statistically significantly increased likelihood of macrolide resistance were not found, possibly due to sample size limitations. The calculation of pharmacokinetic-pharmacodynamic indices found that a longer drug exposure was more closely associated with a successful treatment outcome, but there was not a statistically significant correlation. However, there were few clinical failures in this study giving further credence to the complexity of the in vivo system. There are many factors beyond pharmacokinetics/pharmacodynamics and MICs that contribute to the success of a treatment regimen for cattle suffering from BRD.

Doctor of Philosophy / Department of Anatomy and Physiology / David C. Poole / Exercise-induced pulmonary hemorrhage (EIPH) or epistaxis has been recognized in racehorses since the 16th century. Since this time, great strides have been made in terms of identifying the lungs as the source of the hemorrhage via the endoscope, utilization of bronchoalveolar lavage to quantify the hemorrhage, and the discovery of successful treatments such as furosemide and the nasal strip that ameliorate, but do not abolish EIPH. It has been determined that, in addition to extremely high pulmonary arterial pressures and the negative intrapleural pressures being the major physiologic forces causing pulmonary capillary stress failure, other factors have the potential for influencing the severity of EIPH including locomotory impact trauma, inflammatory airway disease (IAD), upper airway obstruction, coagulation anomalies, and high blood viscosity. It has been hypothesized that EIPH is detrimental to performance and this was recently confirmed by Hinchcliff et al. in 2004. EIPH is a complex multi-factorial condition with much still unknown about the etiology, best method for diagnosis, and most effective form of treatment. Chapter one of this dissertation determined the effectiveness of a novel treatment, concentrated equine serum, in ameliorating EIPH via reduction of IAD. Chapter two refuted the hypothesis that herbal formulations commonly used in the field with anecdotal success would decrease EIPH by correcting coagulation deficits during exercise, as scientific efficacy was not evident, at least at the dose and duration used in our investigation. Chapter three addressed the dogma that EIPH only occurs during maximal intensity exercise, and in demonstrating significant EIPH during sub-maximal exercise, emphasized the role that the airways play in contributing to the initiation and severity of EIPH. Chapter four examined the occurrence and severity of EIPH in the horse’s canine counterpart, the racing Greyhound. The demonstrated presence of mild EIPH in the Greyhound, a physiologically similar yet different athlete in comparison to the horse sheds new light on the etiology of this condition in both species. The results of these investigations have advanced the frontiers of our knowledge concerning EIPH. Specifically, they have generated novel information on the mechanistic bases of EIPH and have provided evidence supporting additional treatment options for reducing the severity of EIPH in horses.

Doctor of Philosophy / Department of Family Studies and Human Services / Anthony Jurich / Until recently, minimal research has been done regarding the impact of veterinary student stress on the student’s significant intimate relationships. In this study of 466 veterinary students enrolled in five different accredited U.S. Colleges of Veterinary Medicine, the association of five primary variables, perceived stress, self-esteem, academic satisfaction, relationship satisfaction, and general life satisfaction was investigated, utilizing McCubbin and Patterson’s (1983) Double ABCX Model as a guide. Six of the seven hypotheses were supported and further regression analysis yielded a model of variable associations that supported the Double ABCX Model, though significant gender differences were found. Specifically, female students perceived significantly more stress than did male students. With the exception of academic satisfaction and relationship satisfaction, all of the major variables were strongly correlated with each other. For the women, who comprised a significant majority of the sample, perceived stress had a significant impact on relationship satisfaction, as well as on self-esteem, which in turn significantly impacted academic satisfaction. Relationship satisfaction and academic satisfaction both significantly, and separately, impacted general life satisfaction, suggesting that relationship satisfaction and academic satisfaction are different constructs that operate independently of each other. However, both have a significant impact upon and are significantly associated with general life satisfaction, suggesting that neither can be neglected during veterinary training if a positive outcome is desired. For the men in this sample, stress impacted directly upon academic satisfaction, self-esteem, and general life satisfaction but did not have a direct impact upon relationship satisfaction. Instead, relationship satisfaction and general life satisfaction had a highly correlated relationship, with both significantly and strongly associated with the other. Although academic satisfaction and relationship satisfaction appear to be two separate entities, they are both important elements to achieving general life satisfaction during veterinary training and, therefore, should be equally attended to during the training process. These findings are interpreted and discussed in light of the implications for and importance of continued collaboration between veterinary students and programs and the field of Marriage and Family Therapy in an effort to understand and support students and their significant relationships during the veterinary training process.

Master of Science / Department of Clinical Sciences / Michael W. Sanderson / A Monte Carlo model was developed to determine the cost-effectiveness of different biosecurity strategies for Bovine Viral Diarrhea Virus (BVDV) on cow-calf farms. Where possible, risk distributions were defined in the course of a critical literature review covering all publications since 1990 relevant to BVDV on cow-calf farms. The prevalence of persistent infections (PIs) in adult cows was unknown, so a survey of viremia in 2,990 adult cows for sale in the Midwest during 2006 was performed; prevalence was calculated to be 0.07%. In order to validate a newly developed RT-nPCR for pooled serum used for the survey, sensitivity was determined based on 100 known viremic serum samples; sensitivity was 95%, with no detectable effect of strain type. A Monte Carlo model was developed to calculate the risk of introducing BVDV to a cow-calf herd and number of PIs introduced in one year, based on herd imports and biosecurity strategies. The results of that model were integrated with a stochastic SIR model for the spread and impact of BVDV through a cow-calf herd over 10 years, based on herd size and control strategies. The resulting model was integrated with a stochastic model for the cost of both the biosecurity and control measures used and the financial impact of BVDV infection on the herd over 10 years. The lowest risk option of 14 biosecurity strategies were calculated for 400-, 100-, and 50-head herds with 8 different import profiles, and management factors that increase financial risk due to BVDV were determined.

Master of Science / Department of Clinical Sciences / Michael Sanderson / Biosecurity, biocontainment and security practices are important in production animal agriculture. Procuring cattle from multiple sources and commingling them into a single confinement operation increases risk of disease introduction. The large concentration of animals makes a feedyard a more likely target of a domestic or international terror group. Controlling or eradicating an intentionally introduced pathogen or toxin would be costly. The aim of these surveys was to gather information from experts about perceived risks and mitigation strategies and to assess current practices of biosecurity, biocontainment and security in Central Plains feedyards. Consulting veterinarians and feedyard managers shared similar views on the likelihood of disease caused by terrorism, natural introduction or accidental introduction, and on the importance of on-site security. They disagreed on the importance of preventative products, disease transmission control, and environmental control. Generally speaking, feedyard managers believed environmental control to be more important than consulting veterinarians. In reference to a survey of current practices, some feedyards use equipment for both manure and feed handling. Many feedyards are not cleaning and disinfecting oral treatment equipment, treatment facilities, or unloading facilities on a regular basis which may increases their risk for indirect disease transmission of endemic agents such as Salmonella or BVDV. Most feedyards in this survey import some cattle directly from an auction market, do not require clean boots or foot covering to be worn by visitors, and do not require trailers to be cleaned. Smaller feedyards were more likely to require trailers to be cleaned before loading incoming cattle. Less than half of the feedyards reported having a fence that will stop humans or kept protein supplements or micro-nutrients secured from access. Some feedyards enforced a visitor log or employed a night watchman. Most feedyards learned about a future employee by calling references listed in resume, but some performed a criminal background check. A cost-benefit analysis should be done on all management practices to determine economic benefits. More research is needed to better understand which practices are most beneficial.

Master of Science / Department of Diagnostic Medicine/Pathobiology / Daniel U. Thomson / Escherichia coli O157:H7 is a human food-borne pathogen and cattle feces are a major source of contamination. Immunization against E. coli O157 may be a practical pre-harvest intervention strategy. A siderophore receptor/porin proteins (SRP) based vaccine has been developed to decrease the prevalence of E. coli O157 in cattle. Two studies were conducted to determine the efficacy of the SRP vaccine. In the first study, thirty calves were randomly assigned to one of two groups: control or SRP vaccine. Two weeks after the second vaccination, calves were orally inoculated with nalidixic acid-resistant (Nal[superscriptR]) E. coli O157. Fecal samples were collected for five weeks. Calves were necropsied on day 35 to collect gut contents and tissue swabs to determine Nal[superscriptR] E. coli O157:H7. The number of calves that were culture positive for E. coli O157 were lower (P= 0.07) in vaccinated group compared to the control. In the second study, cattle in two feedlots were randomized to SRP vaccine or control. Cattle were vaccinated on days 0 and 21. Rectal fecal samples were collected on day 0, and pen floor samples were collected on days 21, 35, and 70. Rectal fecal samples, RAMS, and hide swab samples were collected on d 85. Cattle were weighed on days 0, 21, and 85. Vaccination significantly reduced (P = 0.04) fecal E. coli O157 prevalence. There was also a decrease (P < 0.05) in E. coli O157 prevalence on hides and in fecal samples on day 85 in vaccinated cattle compared to the control.