Center for Veterinary Health Sciences – FY 2005 Research Abstracts
The purpose of this project is to develop new antimycobacterial agents for control and/or eradication of mycobacterial diseases. This project involves collaborations with a Crystallography Group and an Organic Chemistry. This is a drug-design program based upon recombinant DNA technology, X-ray crystallography, organic synthesis, and site-directed mutagenesis.
Sponsor: NIAID/NIH, grant AI-41348
PI: William W. Barrow
Co-PIs: Rebecca J. Morton
SRI, Birmingham, Alabama: William J. Suling, Robert Reynolds
“In Vitro and Animal Models for Emerging Diseases and Biodefense,” Part A: “In Vitro Screens for Antimicrobial Activity”
As directed by the NIH project officer, the College of Veterinary Medicine at Oklahoma State University will develop, validate, and use in vitro assays to screen test substances for activity against emerging “infectious agents.” The activity in this contract will involve the screening of compounds for antimicrobial activity against several biodefense-related select agents in Categories A and B. Materials for testing will be provided by NIAID, as described in RFP NIH-NIAID-DMID-03-39.
Sponsor: NIH, NIAID, DMID
PI: W.W. Barrow
Co-Investigators: K. Clinkenbeard, R. Morton, J. Wyckoff
Narrow–Spectrum Drug Targets for Bacillus anthracis
The goal of this project is to show proof-of-principal for potential antimicrobial targets in a specific biosynthetic pathway of B. anthracis that will allow for the development of compounds that can be used to treat anthrax. The approach will utilize DNA recombinant technology to develop an enzyme assay that will be utilized for high-throughput screening (HTS) of potential inhibitors. A robotics system will be used to implement the HTS. A pharmaceutical company has agreed to supply the project with potential inhibitors which will be screened in the recombinant enzyme assays.
Sponsor: NIAID/NIH, grant R21-AI-055643.
PI: William W. Barrow
A Cell Culture-Derived Vaccine for Anaplasmosis
Anaplasma marginale harvested from tick cell culture will be tested as an antigen for a new and improved vaccine for bovine anaplasmosis. This antigen should result in the development of a vaccine that is safe, easily standardized, and free of contaminating bovine cells and pathogens. A vaccine dose will be formulated and tested in cattle for vaccine efficacy.
Sponsor: Novartis Animal Vaccines, Inc.
PIs: Edmour F. Blouin, Katherine M. Kocan, Jose de la Fuenta
Novartis Animal Vaccines, Inc.: Thomas Halbur, Virginia C. Onet
A Cell Culture-Derived Vaccine for Anaplasmosis
Anaplasma marginale harvested from tick cell culture will be tested as an antigen for a new and improved vaccine for bovine anaplasmosis. This antigen should result in the development of a vaccine that is safe, easily standardized, and free of contaminating bovine cells and pathogens. A vaccine dose will be formulated and tested in cattle for vaccine efficacy.
Sponsor: Oklahoma Center for the Advancement of Science and Technology
PIs: Edmour F. Blouin, Katherine M. Kocan, Jose de la Fuente
Novartis Animal Health: Thomas Halbur, Virginia C. Onet
Anaplasma centrale will be propagated in tick cell culture. Organisms harvested from cell culture will be tested as a live vaccine for bovine anaplasmosis in South Africa and Israel. The cell culture derived A. centrale should result in the development of a vaccine that is safe, easily standardized, and free of contaminating bovine cells and pathogens. A vaccine dose will be formulated and tested in cattle for vaccine efficacy.
Sponsor: U.S.-Israel Cooperative Development Research (CDR) Program
PIs: Edmour F. Blouin, Katherine M. Kocan, José de la Fuente
Kimron Veterinary Institute, Israel: Varda Shkap
Onderstepoort Veterinary Institute, South Africa: Eric Zweygarth
Role of Pasteurella (Mannheimia) haemolytica Leukotoxin in Shipping Fever Pneumonia
Pasteurella haemolytica produces an exotoxin termed leukotoxin (LKT), which has been implicated as an important virulence factor in shipping fever pneumonia in cattle. LKT specifically intoxicates ruminant leukocytes and platelets, but LKT has limited or no effect on leukocytes from other species. Bovine CD18 acts as the species-specific and leukocyte-specific receptor for P. haemolytica LKT. Prior to exposure to LKT, CD18 was found to be evenly distributed around the periphery of the bovine leukocyte plasma membranes. Following one minute of exposure, LKT and CD 18 were found associated with patches of degenerative plasma membranes. No CD18 or LKT was observed in regions of intact plasma membranes. This supports LKT-induced capping of CD18. The rapid degeneration of the associated plasma membrane in these patches suggests that CD18 localization to these regions may play a role in loss of plasma membrane integrity.
Sponsor: Oklahoma Agricultural Experiment Station
PI: Ken Clinkenbeard
The overall goal of this proposal is to develop highly specific approaches for detecting and identifying explosives, nerve gases, and BW agents using a unique amplifying fluorescent polymer (AFP), which will greatly increase the speed and sensitivity of detection of explosives and CBW agents and to apply this development to produce a continuous real-time microarray networkable detection system for use against terrorist threats. Central to the overall goal is the underlying hypothesis that AFP can be used as a sensor platform and functionalized with specific probes to detect in continuous real-time explosives and specific CBW agents at minimal effective levels. This hypothesis was tested for nitroaromatic and other explosives, a nerve gas agent, and three BW agents. The research conducted for this project did the following: 1) develop and tested probes specific and selective for explosives and prototype CBW agents, 2) functionalized AFPs for detection of prototype agents, 3) produced and tested a prototype AFP microarray sensor for explosives and CBW detection, and 4) conducted refinement, application and commercialization of our MIPT AFP microarray sensor.
Sponsors: Oklahoma City Memorial for the Prevention of Terrorism, National Institute of Justice
PIs: Ken Clinkenbeard. CoPIs: Jerry Malayer, Rebecca Morton, John Wyckoff
Commercial Partner: Nomadics, Inc.
The threat from biological warfare (BW) agents in combat and terrorist scenarios makes the ability for rapid detection and identification of BW agents of great importance. Oklahoma State University (OSU) along with our commercial partner, Nomadics, Inc., propose to adapt amplifying fluorescing polymer (AFP) technologies currently being developed for BW agent detection in aerosols for monitoring of water supplies for BW agents. Such a monitor is of great interest to the Joint Services Agent Water Monitor (JSAWM) project of the Army Soldier Biological and Chemical Command (SBCCOM). Hurdles for development of near-real-time continuous monitoring of water supplies for BW agents are requirements for high sensitivity and specificity for up to 20 agents monitored simultaneously with minimal use of consumable reagents. The approach proposed herein is novel using new technologies which can interrogate water supplies with a highly sensitive regenerable reagent adaptable for numerous BW agents. This concept was demonstrated utilizing a prototype portable BW agent water monitor for two BW agents, Francisella tularensis, the causative agent of tularemia, and the water-borne pathogen enterohemorrhagic Escherichia coli O157:H7 (EHEC). Central to the overall goal is the underlying hypothesis that AFP can be used as a sensor platform for BW agents in water supplies and functionalized with specific probes to detect in near-real-time BW agents at minimal infectious levels. This hypothesis was tested for two BW agents. The functionalized AFPs were incorporated into a recoverable microbead (MB) prototype monitor.
Sponsor: ??
PI: Ken Clinkenbeard
CoPIs: Rebecca Morton, Jerry Malayer
Development of Aptamer Beacons to Lipopolysaccharide for the Real-time Sensing of BW Agents
Force protection is of utmost importance, but a lack of real-time sensing technologies for biological warfare (BW) agents leaves U.S. forces vulnerable to conventional as well as terrorist’s BW attacks. The major barriers to real-time sensing of BW agents are: 1) a lack of robust probes for detection of BW agents, and 2) inadequate sensitivity of sensor platforms through which target probes transduce their detection signals. We propose to conduct a “proof-of-concept” project to marry a new type of probe technology with exceptional robustness termed aptamers with a newly conceived sensing platform with exceptional sensitivity termed AFP beacons to overcome the current barriers for developing real-time BW sensing. Aptamers are highly stable and specific oligonucleotides, which work like monoclonal antibodies (Mab) to bind directly to BW target agents. However, unlike Mab, aptamers have exceptional stability, even under field conditions. Aptamer technology is only beginning to be applied to detection of infectious agents. We developed aptamers to surface exposed targets on the prototype BW agent, enterohemorrhagic E coli. We will next demonstrate that these aptamers can be used to specifically detect BW agents. In future experiments we will engineer our anti-E coli aptamers to act as beacons so that when the aptamer binds to its target, it will switch from an “off” to an “on” signal by turning on specific fluorescence directly. No consumable reagents are required. Once the DEPSCoR proof-of-concept project is accomplished, we will transition to prototype sensor development using this technology.
Sponsor: Army Research Laboratory
PI: Ken Clinkenbeard.
Development of Aptamer Beacons for Antemortem Diagnosis of Chronic Wasting Disease
The goal of the project is develop the basic research in support of an aptamer beacon based antemortem diagnostic test for the transmissible spongiform encephalopathy chronic wasting disease (CWD) of elk and deer. Currently available diagnostic tests are based on monoclonal antibodies. These tests are applicable for biopsy or postmortem samples, but may have limitations that preclude their further development as antemortem tests. The limitations of Mab-based tests may be overcome by a new molecular entity known as aptamers. In particular, aptamers can be manipulated using standard molecular biology techniques to act as beacons or molecular switches that turn “on” a fluorescent signal when they bind to their target. Aptamers are nucleic acids that serve as novel recognition molecules or probes that are highly specific for a wide range of targets. Attempts by others to use aptamers to distinguish normal hamster prion from its abnormal isoform were not successful. However, a novel aptamer selection strategy is proposed herein to overcome the problems of the original aptamer selection experiments. We propose to use a crossover selection strategy. Initially, the aptamer selection target will be particular peptides of CWD prion thought to be exposed on abnormal isoform. The reduced aptamer pool will subsequently be selected against the insoluble plaques of the abnormal isoform isolated from brain tissue of affected elk or deer. Aptamer that specifically recognize the abnormal isoform of CWD prion will subsequently be engineered as aptamer beacons—a novel signaling aptamer that have a built-in molecular switch.
Sponsor: U.S. Army Medical Research and Materials Command, National Prion Research Program
PI: Ken Clinkenbeard.
Polymer-Based Yersinia Pestis Point-of-Case Diagnostics
The goal of the proposed research is to develop a highly sensitive and specific multi-locus array diagnostic for rapid identification of Yersinia pestis, the causative agent of plague. This infectious bacterial agent is on the NIAID category A priority list as a potential biological warfare (BW) agent. In order to confirm suspicions that clinical cases may be due to purposeful aerosol dissemination of Y. pestis and, as such, may be indicative of an imminent plague epidemic, and to enable healthcare professionals to instigate immediate and effective therapeutic intervention and control measures, a rapid, sensitive, and accurate method for early detection of disease is a high priority. To meet this diagnostic challenge, the detection technology will use a polymer that exhibits intrinsic amplification of fluorescence transduction events to rapidly identify species-specific genomic, proteomic, and lipo-oligosaccharide (LOS) markers of Y. pestis. This amplifying fluorescent polymer (AFP) will be fabricated as nanoparticles, and functionalized for covalent attachment of quencher-labeled molecular or aptamer beacon probes, which will trigger amplified fluorescent responses when binding of the target to the probe causes dequenching of the polymer. We are confident the amplification afforded by AFP will enable detection of target analytes in extremely low concentrations with minimal sample preparation, thus providing significant advantages over current microbiological and molecular diagnostic methods. Once proof of concept is established using standard microarray technology, the diagnostic platform will be integrated into a real-time, low-density, multi-locus array printed in a membrane sample delivery system that ultimately will be used to identify and discriminate between many different BW pathogens.
Sponsors: National Institute for Allergies and Infectious Disease, Nomadics, Inc.
PIs: Ken Clinkenbeard, Jerry Malayer
Ruminant B-Lymphocyte Yellow Fluorescent Protein Aggregation Bioassay for Elk Chronic Wasting Disease
The goal of the proposed research is to develop a cell culture model for elk chronic wasting disease (CWD) prion propagation that can be used as a bioassay for detecting CWD. CWD is a transmissible spongiform encephalopathy (TSE) caused by the mis-folding of a normal cell surface prion protein (PrPc) through the interaction with infectious mis-folded and protease resistant prion protein (PrPres). The PrPres specific to CWD is PrPcwd. CWD occurs in free ranging and captive elk and deer herds in several Rocky Mountain and Plains states. Although there is evidence suggesting that CWD cannot be transmitted to humans, this potential has not been thoroughly ruled out. Current infectivity bioassays involve the use of live animals, whereas the proposed cell culture model will reduce the need for animal experimentation to study the mechanism of prion infectivity and disease. In addition, a cell culture will be developed as an assay for the screening of infectious prions in veterinary medical samples from elk and deer. This assay system also has the potential for use in assessing therapeutic strategies. Our goal will be accomplished by bioengineering a bovine B-lymphocyte cell (B-cell) line to surface express elk PrPc fused to yellow fluorescent protein (YFP). Like other glycosyl-phosphoinositol (GPI) anchored proteins, surface expressed YFP-PrPc will have a dispersed distribution on these B-cells. Interaction of these B-cells with infectious, mis-folded, protease resistant CWD prion (PrPcwd) will induce conversion of the dispersed YFP-PrPc to aggregated mis-folded YFP-PrPcwd that will be detected by confocal microscopy as aggregated YFP-PrPcwd on the B-cell surface.
Sponsors: US Army Research Office, Nomadics, Inc.
PIs: Ken Clinkenbeard, Jeff Blair
Shipping Fever: New Approaches to Understanding Prevention and Management
The objective is to determine infectious agents and host responses
that cause respiratory disease and/or defend cattle from disease. Naturally
occurring cattle diseases at Noble Foundation, privately owned, USDA, and OSU
herds are being investigated. Emphasis is primarily on bovine viral diarrhea
virus infection and pasteurellosis. Vaccination programs are being evaluated
and new recommendations given.
Sponsor: The
Noble Foundation
PIs: Anthony W. Confer, S. Mady Dabo, Robert W. Fulton, Jerry W. Ritchey
Department of Veterinary Clinical Sciences: John G. Kirkpatrick, Robert A.
Smith
The project determines changing patterns, geographical differences, risk factors, and management practices related to bovine respiratory disease. The influence of various bacteria and viruses is studied. In addition, the pharmacokinetics and efficacy of newer therapies and new generation vaccines are evaluated. The host-pathogen relationship is characterized at the molecular level.
Sponsor: Oklahoma Agricultural Experiment Station
PIs: A. W. Confer, R. W. Fulton, R. J. Panciera, K. D. Clinkenbeard, R. J. Morton
Mannheimia haemolytica Outer Membrane Protein PlpE: Characterization of Epitopes Stimulating Homologous and Heterologous Serotype Protection
This project is a molecular and immunologic approach to studying an immunologically important outer membrane protein of M. haemolytica. It compares the PlpE protein from serotypes 1, 2, and 6 with respect to important epitopes for immunity.
Sponsor: USDA CSREES, National Research Initiative Competitive Grant
PIs: A. W. Confer, Sahlu Ayalew
Mannheimia haemolytica Bacterin-Toxoid Efficacy Studies
This project studies the efficacy of commercial M. haemolytica vaccines in an experimental challenge model.
Sponsor: Pfizer Animal Health
PIs: A. W. Confer, R. J. Panciera
Chimeric Vaccine for Mannheimia haemolytica Infection in Cattle
Recombinant DNA technology is being applied to induce a single chimeric protein that will stimulate immunity in cattle to M. haemolytica leukotoxin and the outer membrane.
Sponsor: Oklahoma Center for the Advancement of Science and Technology (OCAST) Oklahoma Applied Research Program (OARS)
PIs: A. W. Confer, Sahlu Ayalew
Pasteurella multocida OmpA: Functional Characterization
This project studies the adherence and colonization properties of Pasteurella multocida OmpA. It is designed to investigate the specific role of P. multocida OmpA in the bacterium interaction with host cells and in the pathogenesis of the disease.
Sponsor: USDA CSREES, National Research Initiative Competitive Grant
PIs: S. M. Dabo, A.W. Confer
Development of a Vaccine against Ixodes scapularis Infestations
Antigens identified in previous studies to induce protection against Ixodes scapularis tick infestations in mice will be used in vaccine formulations. The vaccine will be tested against the three tick stages: larvae, nymphs, and adults.
Sponsor: Pfizer Animal Health, Inc.
PIs: José de la Fuente, Katherine M. Kocan, Edmour F. Blouin, Consuelo Almazán
The major danger to research and animal care personnel working with rhesus monkeys is monkey B virus (BV), a herpesvirus that is transmitted by bites and scratches and is rapidly fatal in humans if not detected and treated rapidly. This project involves molecular characterization of BV and related viruses of other primates and application of these data to develop more sensitive and specific diagnostic tests that will permit rapid and reliable identification of BV infections in humans.
Sponsor: NIH, NCRR
PI: R. Eberle
Baboons are an important animal species used in biomedical research. This program will develop a breeding colony of baboons in Oklahoma and supports research aimed at improving the breeding efficiency of baboons in captivity, defining viruses that naturally infect baboons, and improving the basic well-being and behavior of captive-bred baboons.
Sponsor: NIH, NCRR
PIs: R. Eberle, A. Kocan, J. d'Offay
University of Oklahoma Health Sciences Center: Gary White
Indigenous viruses can have a major adverse effect on the results of biomedical research studies using animals, particularly where immunosuppression is involved. This program supports derivation of a colony of baboons that are free of all known herpesviruses and most retroviruses.
Sponsor: NIH, NCRR
PIs: R. Eberle
University of Oklahoma Health Sciences Center: Gary White
Bovine Viral Diarrhea Disease Virus (BVDV) Vaccines: Antibody Response to Heterologous BVDV Strains
The study will determine the range of heterologous immunity in calves receiving modified live virus (MLV) or killed BVDV vaccines. Currently there are two recognized antigenic types of BVDV, Type 1 and 2. The study will determine if these vaccines induce antibodies to various Type 1 and 2 viruses.
Sponsors: Grand Laboratories, Inc., Pfizer Animal Health
PIs: Robert W. Fulton, Anthony W. Confer
Genetic and Antigenic Variability of BVDV in Cattle Infections
Bovine viral diarrhea viruses (BVDV) isolates from the Oklahoma Animal Disease Diagnostic laboratory (OADDL) will be obtained from clinical/necropsy cases. The viruses will be typed as BVDV 1a, 1b, or 2. Potentially there will be additional typing and/or groups. Field isolates from naturally occurring disease , including persistently infected (PI) cattle will be compared with vaccinal stains and standard reference strains. A phylogenetic survey of the BVDV subtypes from the field isolates, vaccinal strains, and reference strains will be performed to detect relationships among the virus and their genetic stability. Neutralization tests will be performed to compare the subtypes to the vaccinal strains. Potentially new subtypes may warrant additional subtypes in the vaccines.
Sponsor: Oklahoma Agricultural Experiment Station
PIs: Robert W. Fulton, A. W. Confer
Oklahoma Animal Disease Diagnostic Laboratory: J.T. Saliki
Antibiotic Administration and Vaccination with Live Bacterial Vaccine in Calves
This study will determine if an antibiotic given calves that have been administered avirulent Pasteurella haemolytica and Pasteurella multocida vaccine will decrease the immune responses to the immunogens. Calves will receive Micotil antibiotic and Once PMH Pasteurella haemolytica and P. multocida vaccine. The calves’ sera will be tested for P. haemolytica and P. multocida antibodies.
Sponsor: ELANCO Animal Health, Division of Eli Lilly and Company
PIs: Robert W. Fulton, Anthony W. Confer
Bovine viral diarrhea viruses (BVDV) occur as biotypes, cytopathic (CP), and noncytopathic (NCP), and as genotypes 1 and 2. Certain BVDV disease forms occur with different biotypes/genotypes. The molecular differences among biotypes/genotypes will be investigated by PCR and nucleic acid sequencing. Virulence markers of BVDV will be investigated.
Sponsor: Oklahoma Agricultural Experiment Station
PIs: Robert W. Fulton, Jean M. d’Offay, Anthony W. Confer, Jerry W. Ritchey
Oklahoma Animal Disease Diagnostic Laboratory: Jeremiah T. Saliki
Evaluation of Viral Vaccine Containing Infectious Bovine Rhinotracheitis Virus (IBRV), Bovine Viral Diarrhea Virus 1 and 2 (BVDV), Parainfluenza -3V (PI-3V), and Bovine Respiratory Syncytial Virus (BRSV) in Preventing Infection and Respiratory Disease in Cattle
The purpose of the study will be to determine if pre-weaning vaccination of ranch calves with viral vaccine: 1) reduces respiratory disease, and 2) reduces transmission of viruses in calves moved from auction markets and commingled with the fresh calves under feedlot conditions.
Sponsor: Fort Dodge Animal Health
PIs: Robert Fulton, A.W. Confer
Oklahoma Animal Disease Diagnostic Laboratory: J.T. Saliki
Department of Veterinary Clinical Sciences: D.L. Step
Vaccination of Ranch Calves with Modified Live Viral Vaccine: Effects on Viral Transmission on Commingled and Transported Calves
This study will determine if vaccination of ranch calves with a modified live viral vaccine will reduce respiratory disease and reduce transmission of viruses in the calves commingled with auction calves and moved to a feedlot.
Sponsor: Schering Plough Animal Health Corp.
PIs: Robert W. Fulton, Anthony W. Confer
Food Safety: Farm to Table
The long-term objectives are to develop methods for assuring the microbial safety of the food supply from farm to table. Focus is on control of Salmonella species and Escherichia coli O157:H7 associated with swine and cattle.
Sponsor: USDA/CSREES
PIs: Terry Lehenbauer
College of Agricultural Sciences and Natural Resources: Stanley Gilliland, Guolong Zhang, Peter Muriana, Siobhan Reilly
Diagnostic Assays for Borrelia lonestari
This project supports development of diagnostic assays for Borrelia lonestari to use in human patients, wildlife reservoirs, and tick vectors. This grant will advance our ability to identify infection with novel Borrelia spp. and investigate its role as an agent of human disease.
Sponsor: NIH/NIAID
PI: Susan E. Little
Prototype FSU Sensor Testing
Sensor technologies that have been developed in the former Soviet Union to detect biological threat agents are being evaluated for potential use in a fully integrated warning and response system to protect individuals, property, and the environment from the threats of accidental or terrorist release of biological agents. Our laboratory is involved in the selection, developmental assessment, and testing of such prototype detection systems.
Sponsor: General Atomics
PI: Rebecca J. Morton
Murine Model for Monkey B Virus Infection
Non-human primates are an integral part of biomedical research programs, particularly as animal models of human disease. As long as monkeys are used for research, zoonotic infection by simian viruses will continue to be an occupational hazard. Human infections by monkey B virus, a herpes virus of macaques, involve the central nervous system and are usually fatal if left untreated. The goal of this project is to establish and characterize, both pathologically and immunologically, a murine model for B virus infections. Establishment of a well-characterized murine model system for B virus infection will serve as an important resource in which hypothesis-driven studies can be performed, such as evaluation of anti-viral drugs, challenge assessment of potential vaccines, and investigation of the role of various viral genes/proteins in determining the pathogenic properties of these viruses.
Sponsor: National Institutes of Health
PI: Jerry W. Ritchey
A Study of the Viral, Bacterial, Mycologic, and Toxicologic Conditions Associated with Marine Mammal Strandings in the Gulf Coast of Mississippi and the Central Gulf Coast of Mexico
This is a comprehensive two-year study aimed at identifying the causes of stranding and death among dolphins in the Gulf coast of Mississippi. Viruses, bacteria, fungi, and toxins affecting dolphins in this region will be determined and detailed pathological lesions from dead dolphins will be described.
Sponsor: National Marine Fisheries Service (Prescott Grant Program)
PIs: Jeremiah T Saliki, Uriel Blas-Machado, Sandra Morgan, Ronald D Welsh
Institute of Marine Mammal Studies: Moby A. Solangi
On Watch for West Nile Virus in Oklahoma
This project aims to detect West Nile virus (WNV) in wild birds and use the information to assess the risk of West Nile to human health in various counties. There is an established correlation between intensity of WNV activity in wild birds and increased risk of human WNV infections.
Sponsor: Oklahoma State Department of Health
PI: Jeremiah T. Saliki
Functional Genomic/Proteomic Analysis of Campylobacter spp.
Campylobacter is the most common agent of gastroenteritis infection in humans. Despite an intensive research effort on understanding Campylobacter pathogenesis, conclusions on the exact mechanism of infection are extremely difficult to draw. This project proposes to investigate the mechanism of Campylobacter pathogenesis using functional genomic tools.
Sponsor: National Institutes of Health
PIs: Alain Stintzi
University of Oklahoma Health Science Center: John Iandolo
Response of Campylobacter jejuni to Host Temperature
The major goals of this project are to study the mechanism of C. jejuni adaptation to various host temperatures and characterize the interaction of C. jejuni with the chicken gastrointestinal tract. This work will shed some light on the biology of bacterial adaptation to temperature changes and its role in the colonization and infection processes.
Sponsor: Oklahoma Center for the Advancement of Science and Technology
PI: Alain Stintzi
Campylobacter Colonization and Virulence Determinants
The major goal of this project is to develop and use novel functional genomic tools to identify some of the Campylobacter colonization and virulence determinants. The identification of these in vivo Campylobacter determinants could significantly contribute to the development of more effective methods to diagnose, manage, and ultimately prevent Campylobacter infections.
Sponsor: National Institutes of Health
PI: Alain Stintzi
Cytokine Expression in Response to Brucella Vaccines
Cytokine responses induced by Brucella polysaccharides in blood cells from vaccinated and unvaccinated cattle and mice, unvaccinated pigs, and humans were evaluated in vitro. Reverse transcriptase-polymerase chain reaction was used to detect cytokine mRNA. Development of a cytokine diagnostic profile indicative of induction of immunity for application to human vaccinates was attempted.
Sponsor: Canada Department of National Defense
PI: John H. Wyckoff III
Synthetic peptides with sequences homologous to Mycobacterium bovis proteins will be used to develop better test reagents for field and laboratory diagnosis of bovine tuberculosis. Cattle immunized with M. bovis will be compared for responsiveness to these antigens and those of M. avium to determine both specific and cross-reactive responses.
Sponsor: Oklahoma Agricultural Experiment Station
PIs: John H. Wyckoff III
DEPARTMENT OF PHYSIOLOGICAL SCIENCES
The major goals of this project are to elucidate human sulfotransferase (SULT) chemical and kinetic mechanisms, to understand physiologic functions of SULTs, and to investigate their relevance to human health in physiologic and pathologic conditions. Research focuses on: 1) mechanisms of enzyme catalysis, substrate inhibition, and product activation of human SULTs; 2) effect of clinical widely used drugs on human sulfotransferase catalytic activities; and (3) oxidative regulation mechanisms of human SULTs.
Sponsors: National Institute of Health, Oklahoma State University
PI: Guangping Chen
Studies in this project focus on Sulfotransferases (SULTs) induction and their induction mechanisms. Rats, human hepatic carcinoma cell line, Hep G2, and human intestinal carcinoma cell line, Caco-2, will be used for these studies. Enzyme activity assay, Western blot, reverse transcription polymerase chain reaction (RT-PCR), site-directed mutagenesis, plasmid transfection, small interfering RNA (siRNA) gene silencing, promoter gene deletion, DNA footprinting, and electrophoretic mobility shift assay will be used to determine the induction of SULTs and nuclear receptor mediated SULT induction mechanisms.
Sponsor: Oklahoma State University
PI: Guangping Chen
The long-term goal of the project is to evaluate the contribution of the mar regulon to survival of Salmonella serovar Typhimurium in vivo, in the absence of antibiotics or in the presence of sub-therapeutic and the therapeutic levels of antibiotics.
Sponsor: OCAST Health Research
PI: Cyril Clarke
The objective of this research is to develop a novel detection system utilizing an amplifying fluorescent polymer for rapid, highly-sensitive, and selective detection of methicillin-resistant Staphylococcus aureus present in clinical exudates without prior culture and isolation of bacteria.
Sponsors: Nomadics, Inc, National Institutes of Health
PIs: Cyril Clarke, Jerry R. Malayer
Exercise-Induced Airway Injury in Horses
This project has demonstrated that horses damage the lining of their airways during cold weather strenuous exercise. This injury leads to an asthma-like syndrome of airway inflammation and bronchoconstriction that may adversely affect the horse’s exercise ability. Furthermore, this project has demonstrated that exercise while breathing cold air causes increased expression of mediators that may increase the subject’s susceptibility to viral respiratory disease, thus possibly explaining why athletes have increased frequency of colds and flu after competition and further explaining why the prevalence of these diseases is higher during the winter.
Sponsors: United States Department of Agriculture, Oklahoma Agricultural Experiment Station, Thoroughbred Charities of America, National Institutes of Health
PIs: Michael S. Davis, Jerry Malayer, Katherine Williamson
Animal Health Trust, Newmarket, United Kingdom: David Marlin
Department of Statistics: Mark Payton
Metabolic Strategies and Consequences of Extended Endurance Exercise Fatigue
Metabolic strategies and consequences of extended endurance exercise fatigue resulting from prolonged activity is a leading contributor to battlefield morbidity and mortality. This project focuses on racing sled dogs, which have a four-fold greater capacity for sustained exercise than the best trained human athlete, to determine the metabolic strategies employed by the sled dogs to permit such fatigue resistance. Despite the relatively greater exercise capacity, sled dogs suffer from many of the same exercise-induced diseases as human athletes. Thus, a second goal of this project is to determine the specific causes of these diseases and test strategies to prevent them.
Sponsors: National Institutes of Health, Defense Advanced Research Projects Administration
PIs: Michael S. Davis, Katherine Williamson
Texas A&M University: Michael Willard
Ohio State University: Kenneth Hinchcliff
Molecular
Mechanisms of Lung Surfactant Secretion
Lung surfactant is a surface-active material that stabilizes alveoli. It is
synthesized and secreted by lung epithelial type II cells. The long-term
objective of this proposed project is to elucidate the molecular mechanisms of
lung surfactant secretion from type II cells, including transport of secretory
granules to and fusion with the plasma membrane. Deficiency of lung surfactant
is the cause of respiratory distress syndrome in premature infants.
Accomplishing the goals of this proposal may give a valuable insight to the
therapy of pulmonary diseases such as neonatal respiratory distress syndrome.
Sponsor: National Institutes of Health
PI: Lin Liu
Mechanisms of Alveolar Epithelial Cell Differentiation
The goal of this grant is to understand molecular mechanisms of the differentiation of alveolar epithelial cells in isolated alveolar epithelial cells, fetal lung development, repair of injured lungs using in-house made 10K rat gene arrays, and RNA interference.
Sponsor: National Institutes of Health
PI: Lin Liu
GABA Receptor and Pulmonary Fluid Transport
The major goal of this project is to identify GABA receptor in type II cells as it relates to chloride secretion.
Sponsor: March of Dimes Birth Defects
PI: Lin Liu
Quantitative DNA Microarray: Application in Cardiopulmonary Diseases
The goal of this pre-doctoral fellowship is to develop a quantitative DNA microarray.
Sponsor: American Heart Association
PI: Zhongming Chen (Mentor: Lin Liu)
Characterization and Functional Studies of GABA-A Receptors in Alveolar Epithelial Cells
The goal of this pre-doctoral fellowship is to characterize GABA-A receptors and study their functions in lung epithelial cells.
Sponsor: American Heart Association
PI: Nili Jin (Mentor: Lin Liu)
Nuclear
Receptor Gene Expression in the Bovine Pre-implantation Embryo Produced In
Vitro
Objectives include the isolation of estrogen and retinoic acid-responsive gene
products in ruminants from the developing fetal reproductive tract, adult
female reproductive tract, and pre-implantation embryo.
Sponsor: Oklahoma Agricultural Experiment Station
PI: Jerry R. Malayer
Biological
Point Detection Based on the Amplifying Fluorescent Polymer Platform
Investigators from
the College of Veterinary Medicine and Nomadics, Inc. are working to use
amplifying fluorescent polymer (AFP) films and particles to improve the speed
and sensitivity of nucleic acid hybridization-based detection assays by several
orders of magnitude. This will directly impact detection of biological
materials in the environment by improving sensitivity.
Sponsors: U.S. Army, Nomadics, Inc.
PI: Jerry R. Malayer
Electron Microscopy Laboratory
The Electron Microscopy Laboratory provides instruction in electron microscopy at the graduate level and offers a wide range of microscopy services, including sample preparation, ultrathin sectioning, transmission and scanning electron microscopy, X-ray microanalysis, and confocal microscopy. Researchers may have all of the work done by lab personnel or do some or all of it themselves using lab equipment.
Sponsor: Oklahoma State University
PIs: Charlotte L. Ownby, Phoebe Doss, Terry Colberg
Purchase of Analytical Transmission Electron Microscope and Cryoultramicrotome
This project provides funding for a new 120KV Transmission Electron Microscope with scanning attachment, x-ray analysis and digital imaging capability. It also provides funds for a cryoultramicrotome with which to cut frozen thin sections. The cryoultramicrotome will be equipped with a video monitor to assist in teaching students to make ultrathin sections.
Sponsor: National Science Foundation
PIs: Charlotte L. Ownby, Allen W. Apblett, Bret N. Flanders, Warren T. Ford, Jeanmarie Verchot
Presynaptic Modulation of Anticholinesterase Toxicity
The project evaluates contribution of presynaptic neurochemical mechanisms (e.g., in particular neurotransmitter release) in differential toxicity of organophosphorus insecticides.
Sponsor: National Institute of Environmental Health Sciences
PIs: Carey Pope, Jing Liu Pope, Guangping Chen
Water-Soluble Nanoparticles for Oxygen Imaging in Tumor
Researchers from the Center for Veterinary Health Sciences and Nomadics, Inc. are evaluating the ability of nanomaterials to enhance tumor imaging. Both applications and toxic potential are jointly being investigated.
Sponsors: National Cancer Institute, Nomadics, Inc.
PIs: Carey Pope, Guangping Chen
Nanoparticle Self-Lighting Photodynamic Therapy for Ovarian Cancer Treatment
Collaborative studies between researchers at the Center for Veterinary Health Sciences and Nomadics, Inc. are evaluating the potential for nanoparticles coupled to radical generators to be used in photodynamic therapy for cancer. Feasibility and toxicity studies are concurrently conducted.
Sponsors: U.S. Army, Nomadics, Inc.
PIs: Carey Pope, Jing Liu Pope, Guangping Chen
DEPARTMENT OF VETERINARY CLINICAL SCIENCES
Since the establishment of the Biomedical Laser Laboratory within the College of Veterinary Medicine, research to establish protocols for clinical applications in veterinary medicine has been a primary objective. In addition, the use of laboratory models has resulted in transfer of technology to both industry and human medicine. Work will continue concentrating on the clinical applications of biomedical lasers coupled with collaborative research protocols involving basic scientists (engineers, physicists) and clinicians.
Sponsors: McCasland Foundation, the Mercy Works Foundation
PI: Kenneth E. Bartels
Photodynamic Therapy in Combination with Chemotherapy and Immunotherapy for Treatment of Metastatic Mammary Tumors
Photodynamic therapy (PDT) can be an effective means for direct tumor destruction through its selective photochemical reaction. Common sensitizers used in PDT are 5-aminolevulinic acid (ALA) and Photofrin®. By itself, PDT using ALA and Photofrin® can be effective in some cases, but not effective in others. To address this inconsistency, combinations of several drugs can result in synergistic or additive effects for certain types of cancer such, as mammary carcinoma. Combining multiple tumor treatment modalities may create a type of “cocktail therapy” that could be successful for primary and metastatic disease caused by some types of cancer in both animals and human beings. It is hypothesized that the combined use of PDT and the glycated chitosan (GC) immunoadjuvant may significantly improve the efficacy of the treatment, particularly in treating metastatic tumors. It is further hypothesized that the concurrent use of PDT and a new chemotherapeutic agent, AEADA (Bis-1, 4 aminoethylamino-5, 8-dihydroxyanthraquinone2HCl), may enhance the systemic effect of cancer treatment. Combinations of these treatment modalities will be evaluated in the rat and mouse mammary tumor models.
Sponsor: Kleberg and Barbour Foundations
PIs: Kenneth E. Bartels
University of Central Oklahoma: Wei Chen
Determination of Bloodflow of Aqueous Solutions following Intramuscular Injection in the Green Iguana (Iguana iguana)
Reptiles have renal portal systems that conduct substances injected intravenously in the caudal portion of the body directly to the kidneys, thus possessing an increased risk of organ damage with nephrotoxic substances. This study was undertaken to determine if the same was true for intramuscular injections using nuclear scintigraphy in a green iguana model.
Sponsor: College of Veterinary Medicine
PIs: Armando Burgos, John P Hoover, Tawnia Zollinger
Pharmacokinetics of Single Dose, Topical Silamectin in America Bullfrogs (Rana catesbeiana)
Internal parasites are an important health management problem for captive amphibians. This study was undertaken to determine the pharmacokinetics of a single dose of the antiparasitic drug silamectin applied topical at an empirical dose in a bullfrog model and to assess if there were local or systemic adverse effects.
Sponsors: Oklahoma City Zoo, College of Veterinary Medicine
PIs: Jennifer D’Agostino, John P Hoover
Oklahoma City Zoo: Gary West
Laser Ablation and Cryoepilation as Alternatives to Surgical Pinioning in a Pigeon (Columba livia) Model
The traditional de-flighting techniques for birds that employ surgical alteration or amputation are not well suited for exotic pets or avians that are on display. The diode laser and cryoprobe were evaluated as methods to ablate primary feather follicles as an alternative to these surgical de-flighting techniques in birds.
Sponsor: College of Veterinary Medicine, Laser Laboratory
PIs: Jennifer J. D’Agostino, Timothy Snider, John Hoover
Oklahoma City Zoo: Gary West
Thyroid Hormone Levels in Captive Galapagos Tortoises (Geochelone elephantopus)
Hypothyroidism has been suspected in Galapagos tortoises that develop non-pitting myedema of the head and neck and other extremities. This study was undertaken to determine the thyroid hormone levels of affected and normal Galapagos tortoises.
Sponsor: Oklahoma City Zoo
PIs: Cynthia L DiGesualdo, John P Hoover
Oklahoma City Zoo: Gary West
Oxford Biomedical Research: Thomas R Brown
Urea Space and Body Condition Score to Predict Body Composition of Meat Goats
Yearling Boer x Spanish wethers (40) were used to develop and compare body composition prediction equations for mature meat goats based on urea space (US) and body condition score (BCS). After determining FBW and US, feed and water were withheld for 24 hours before US determination. A 20% (weight/volume) urea solution in 0.9% saline was infused into the jugular vein with a winged catheter (19 gauge, 1.9 cm; Vacutainer®, Becton Dickinson, Rutherford, NJ) at 130 mg/kg FBW after a 0-min sample was collected. Twelve minutes later another sample was withdrawn. Thereafter, shrunk BW (SBW) was determined and blood was placed in ice then centrifuged later. Serum was frozen until analysis of urea (Chaney and Marbach, 1962). US was calculated as urea infused/change in serum concentration.
Sponsors: College of Veterinary Medicine, Langston University
PIs: Lionel J. Dawson. T. Ngwa, R. Puchala, G. Detweiler, R.C. Merkel, I. Tovar-Luna, T. Sahlu, A.L. Goetsch
Nutrient Requirements of Goats: Composition of Tissue Gain and Loss by Mature Goats
The overall objective of the study was to determine the composition of tissue loss and gain by mature goats that varied in initial body weight (BW) and body condition score (BCS). The initial BW and BCS were achieved by subjecting mature Boar x Spanish goats to a management scheme that elicited two levels (thin and fat) of body constitution or fatness prior to the onset of the experiment. A secondary objective was aimed at developing equations that will enable the prediction of body composition of mature meat goats from shrunk body weight after urea dilution. The third objective was to characterize the effects of body condition and nutritional plane on energy metabolism.
Sponsors: College of Veterinary Medicine, USDA
PIs: Lionel J. Dawson, A.T. Ngwa, A.L. Goetsch, R. Puchala, G. Detweiller, T. Sahlu, R. Merkel
Enhanced Goat Production Systems for the Southern United States
Overall project goals are to improve the compatibility of goat production systems with available resources throughout the U.S. Supporting objectives are: 1) develop a vehicle to appraise compatibility of available resources and production conditions with goat production systems; 2) project most appropriate goat production systems based on compatibility with presently available resources and production conditions, and evaluate changes in resources or production conditions necessary for employment of alternative, preferred systems; and 3) disseminate and provide training in use of the developed decision-support vehicle.
Sponsors: College of Veterinary Medicine, USDA, Langston University
PIs: Lionel J. Dawson
Training and Research in Ethiopia
This project is a partnership designed to improve the research, extension, and teaching capability of Alemaya University. The goal was accomplished through a program of training, research, and a development project that targeted household food security, income generating potential, and women in development through the provision of goats and production training to women smallholders.
Sponsors: College of Veterinary Medicine, Langston University, Alemaya University, United Negro College Fund Special Programs
PIs: Lionel J. Dawson
Training and Research in Iraq
The American Institute for Goat Research collaborated in a University of Oklahoma-led project with Oklahoma State University and Cameron University. Other collaborating institutions were Salahaddin University, Basrah University, Al-Anbar University of Technology, and Babylon University in Iraq. The role of the Institute and Langston University was to provide training in small ruminant production to Iraqi scientists and establish a ruminant nutrition laboratory at Salahaddin University.
Sponsors: College of Veterinary Medicine, Cameron University
PIs: Lionel J. Dawson
Introduction and Rationale
Gastrointestinal tract parasitism causes a significant economical loss in goat production. It is known that nutrition status, especially protein intake, affects the establishment of parasites in the gastrointestinal tract. Compared to those fed a straw-based diet, sheep fed a Lucerne-based diet harbored less nematodes at 56 days after infection.
Sponsors: College of Veterinary Medicine, Langston University
PIs: Lionel J. Dawson
Evaluation of Tepoxalin in Reducing Intraocular Inflammation in the Dog
Intraocular inflammation is a common ophthalmic problem in the dog, and is seen following routine cataract surgery. Several non-steroidal, oral anti-inflammatory medications are now available for dogs. Tepoxalin is unique in that it targets both the lipoxygenase and cyclooxygenase pathways of inflammatory mediators. This study uses an aqueocentesis model of intraocular inflammation to evaluate the effect oral tepoxalin has on decreasing inflammation.
Sponsors: OSU Department of Veterinary Clinical Sciences, Schering-Plough Corporation
PI: Margi Gilmour
Normal Blood Thiamine Levels of African Lions (Panthera leo) as Fed in North American Zoos
Thiamine (Vitamin B1) deficiency may occur in large cats that are fed inappropriate diets resulting in neurologic signs that may include ataxia and seizure-like episodes. This study was undertaken to establish reference values for African lions held in North American zoos that were eating appropriate diets.
Sponsor: Department of Veterinary Clinical Sciences
PIs: John P Hoover, Cynthia L DiGesualdo
Evaluation of Bacterial Contamination following Uncomplicated Equine Castrations
Elective castrations were performed on 30 horses by fourth year veterinary students supervised by a clinician. Cultures were obtained from each hemi-scrotum at the conclusion of surgery, following recovery from anesthesia, and at eight and 24 hours post-operatively. From the 240 samples obtained, 350 bacterial isolates were identified.
Sponsors: Biggs-Oxley Chair, Equine Sports Medicine
PIs: H. David Moll, Mike Schoonover
Tissue Adhesion Characteristics of a Textured Intravascular Device
Infection of long-term catheters at the skin surface can lead to numerous complications and greater expense. Increased adherence of the catheter to the skin and subcutaneous tissue may reduce infection rates. A pig model was used to investigate the effect of catheter surface texture on catheter-tissue adhesion properties.
Sponsor: Medical Inventions, LLC
PIs: Mark Rochat
Center for Veterinary Health Sciences: Jerry Ritchie, Charlotte Ownby
Statistics: Mark Payton
University of Central Oklahoma: Wei Chen
Comparison of Two Tonometers (Tonopen-Vet and Tonovet) in Several Species
Glaucoma is a common and serious ophthalmic disease in animals. To determine if an animal has glaucoma, the intraocular pressure must be measured using a tonometer. The current standard tonometer is the Tonopen-Vet, which is widely used in veterinary medicine. This study was undertaken to compare the Tonopen-Vet with a new tonometer, the Tonovet, in horses, cattle, goats, rabbits, alpacas, dogs, and cats. The Tonovet uses a different technology for pressure measurement. The study evaluates the difference in values between both tonometers and the ease of use of each tonometer,
Sponsor: OSU Department of Veterinary Clinical Sciences
PIs: Jeff Studer, Margi Gilmour
The Effect of Gastrointestinal Blood on Plasma Uric Acid, Creatinine, and Urea Nitrogen Values in a Domestic Pigeon (Columba livia) Model
Increases in serum urea nitrogen are seen primarily with gastrointestinal hemorrhage in mammals. This study was undertaken to determine if similar increases in nitrogenous waste products, most importantly uric acid, in avian serum, occurs in non-carnivorous species using a domestic pigeon model.
Sponsor: Department of Veterinary Clinical Sciences
PIs: Julie Sheldon, John P. Hoover
Pathologic Changes Associated with Injectable Carprofen in Pigeons (Columba livia)
Post operative pain management has become an important consideration in avian species. This study was undertaken to determine if there were any pathologic changes on necropsy and histologic examination following single and repeated injections of the nonsteroidal analgesic caprofen at an empirical dose.
Sponsor: College of Veterinary Medicine
PIs: Tawnia Zollinger, John P. Hoover, Armando Burgos
OKLAHOMA ANIMAL DISEASE DIAGNOSTIC LABORATORY
Oklahoma Animal Disease Diagnostic Laboratory
The Oklahoma Animal Disease Diagnostic Laboratory provides accessible and accountable diagnostic service for Oklahoma veterinarians and animal owners in all 77 counties. Early detection of diseases provides the starting point for reducing their incidence and threat. The Laboratory also acts as a frontline sentinel for new and emerging diseases. OADDL promotes and protects the health and economic welfare of Oklahomans, supports the teaching and research missions of the OSU College of Veterinary Medicine, and conducts self-supported research aimed at developing more precise test procedures for commonly encountered, as well as emerging and foreign animal diseases that may produce catastrophic losses (e.g., bovine viral diarrhea, malignant catarrhal fever, parvovirus disease, avian influenza, equine viral arteritis and encephalitis, and toxicoses related to oilfield wastes and agricultural chemicals). The Laboratory conducts research and diagnostic tests for morbillivirus infections of marine mammals. The Laboratory, in conjunction with the Oklahoma State Department of Health, is involved with surveillance of West Nile fever in Oklahoma horses and birds. The Laboratory maintains full accreditation by the American Association of Veterinary Laboratory Diagnosticians.
PIs: Bill J. Johnson and staff