2013 Summer Scholars Faculty Research Projects
Project categories:
Comparative Medicine & Biology
Project Title: Osteoarthritis severity in MIF-KO and CD74-KO mice
Mentor: Dr. Cathy Carlson
Contact: carls099@umn.edu
Co-investigators: Dr. Richard Loeser and Dr. Richard Miller
Project Description: Osteoarthritis (OA) is an important age-related disease, but the mechanisms for its development are not fully understood and no treatments currently exist to stop or slow disease progression. Macrophage migration inhibitory factor (MIF) is a pro-inflammatory cytokine produced by chondrocytes. The absence of MIF has recently been associated with increased lifespan in mice and with a significant reduction in OA severity in adult mice. In the present study, stifle joints of aged MIF- and CD74- (CD74 is the MIF receptor) knockout mice and their corresponding age-matched controls will be evaluated. Histological sections through the entire joint will be subjected to a histological grading scheme and histomorphometry measurements of joint tissues will be taken and analyzed. Although the stifle joints of MIF-KO mice have been evaluated previously, these were from 1- and 2-year-old mice and not mice (as in the present study) that were at the end of their natural lifespan. OA has not previously been examined in CD74-knockout mice.
Techniques and Applications the Student will Learn: Embedding, sectioning, and staining of paraffin-embedded sections of decalcified joint tissues, subjective grading of lesions of osteoarthritis in joint tissues, histomorphometric evaluation of joint tissues, data analysis, abstract preparation and presentation Eligible Funding Sources: NIH/CVM/Merial Back to top Project Title: Fate of Neural Stem cells during viral encephalitis
Mentor: Dr. Maxim Cheeran
Contact: cheeran@umn.edu
Co-investigator: Dr. Walter Low, Department of Neurosurgery
Project Description: Herpes simplex virus (HSV)-1 brain infection is the most common cause for sporadic encephalitis in the United States, which is fatal when untreated. Unfortunately, despite effective acyclovir therapy, majority of surviving patients will suffer long-term neurological impairment. Our laboratory studies a population of cells in the brain called neural stem cells (NSCs), which possess the ability to repair damaged brain tissue. NSCs utilize inflammatory cues to direct the reparative processes in the brain. However, to date nothing is known about viral-induced cues that activate NSCs. Two summer research projects are available in our lab for students interested in research on neuroimmune responses to HSV-1 brain infection. The projects are (1) to investigate if NSCs migrate to sites of HSV-1 induced brain damage in response to inflammatory cues generated by T cells and (2) to study the immunosuppressive effects of NSCs on CD8 T cell response during herpes encephalitis. We will utilize a well characterized mouse model of HSV-1 encephalitis to test the hypothesis that modulating neuroinflammatory responses to HSV-1 infection will foster NSC reparative properties. These are interdisciplinary studies, providing exposure to three major scientific disciplines, including virology, immunology, and neuroscience. The ultimate goal of this research is to develop a new understanding of interactions between NSCs and the immune milieu generated during viral encephalitis that will lead to new therapies for brain damage ensuing virus infection.
Techniques and Applications the Student will Learn: Stem cell culture, Flow cytometry, cryosectioning and immunohistochemistry skills. BSL-2 procedures, involved in handling viruses and animals infection. Assist in performing intracranial surgery in mice to transplant cultured NSCs. Eligible Funding Sources: NIH/SDEC/VPM
Back to top Project Title: Critical role of ROS in the median preoptic nucleus in the development of heart failure.
Mentor: Dr. John Collister
Contact: colli066@umn.edu
Co-investigator: N/A
Project Description: Coronary artery disease leading to myocardial ischemia and infarction (MI) is the primary cause of chronic heart failure (HF). Despite advances in the diagnosis and treatment of heart disease, MI induced HF continues to be a leading cause of morbidity and mortality in the United States. In this application we will use the rat model of coronary artery ligation to create a myocardial infarct and eventual heart failure. It is well known that the renin angiotensin (RAS) is activated and produces much of the pathophysiology of HF. It is also well known that increased RAS in the central nervous system contributes to HF, although the exact locations are not known. Additionally, it has been shown that excessive reactive oxygen species (ROS) in the brain contribute to the pathophysiology of post MI HF in the rat in part due to increased central nervous system RAS activity. In this application we will specifically examine the role of ROS in the median preoptic nucleus (MnPO), a site known to be sensitive to both increased RAS and ROS, in the progression and pathophysiology of MI induced HF in the rat. It is likely that ROS, such as O2•-, in the MnPO are important mediators in the central mechanisms mediating the pathophysiology of HF and therefore we will examine the role of MnPO O2•- in the pathophysiology of HF to address the following Hypothesis: Elevated O2•- in the MnPO contributes to the pathophysiology of HF and decreased cardiac function.
Techniques and Applications the Student will Learn: Continuous measurements will be made of blood pressure and heart rate, as well as sodium/water balance in myocardial infarcted rats previously subjected to injection of the MnPO with adenoviruses capable of overexpressing superoxide dismutase (SOD) (for ROS scavenging). Cardiac function will be assessed at 0, 2, and 4 weeks post MI via echocardiography. Students will have the opportunity to learn techniques related to whole animal physiological chronic experimentation including central injection of adenoviral vectors capable of overexpression of SOD, MI induced HF in the rat, and assist in echocardiography of post-MI rats. These novel studies will add to our understanding of the overall central regulation and homeostatic changes occurring in the post MI induced HF model in the rat. Eligible Funding Sources: NIH/CVM/Merial Back to top Project Title: Lysosomal drug sequestration as a drug resistance mechanism in canine hemangiosarcoma
Mentor: Dr. Erin Dickerson
Contact: edickers@umn.edu
Co-investigator: N/A
Project Description: Canine hemangiosarcomas are highly aggressive tumors in dogs that are largely drug resistant. As a result, the response to treatment is unrewarding causing these tumors to be essentially incurable using conventional therapies. We recently discovered that high amounts of small molecular inhibitors and chemotherapy agents appear to be sequestered in extremely large amounts within intracellular organelles, most likely lysosomes. Sequestration of drugs within the lysosomal space would reduce the amount of drug available and prevent drugs from getting to their targets in sufficient amounts. This high capacity for drug sequestration into lysosomes/lysosomal-related organelles may be a primary and as yet unidentified mechanism for the observed drug resistance in these tumors. For this project, the Summer Scholar will determine if drug sequestration occurs via transport through the late endosomal/lysosomal pathway using a series of fluorescent markers, as well as observing drug sequestration in these organelles under different treatment conditions. In addition, the Scholar will determine if drug sequestration can be inhibited by disruption of lysosomal function, potentially restoring drug distribution. A better understanding of this drug resistance mechanism could lead to improved clinical application and practice.
Techniques and Applications the Student will Learn: Fluorescence labeling techniques/immunohistochemistry; confocal microscopy, cell culture, immunoblotting, cell viability assays; drug treatment assays. The student will learn hypothesis testing, experimental design, laboratory notebook protocols, record keeping, data presentation. Eligible Funding Sources: NIH/CVM/Merical/Oncology Fellow Back to top Project Title: Survival and regeneration of beta cells in diabetes
Mentor: Dr. Meri Firpo
Contact: firpo001@umn.edu
Co-investigator: N/A
Project Description: Diabetes is characterized by high blood sugar as a result of either insufficient insulin production by the pancreatic beta cells (type 1) or a failure of the body to utilize the insulin produced (type 2). In both types of diabetes, a loss of beta cells occurs over time, either from oxidative stress and inflammation through programmed death, or apoptosis, or direct immune killing for type 1 diabetes. Recently, however, oxidative stress has been identified as a mediator of immune killing of cells. This means that strategies developed for the protection of beta cells from apoptosis in type 2 diabetes may similarly protect beta cells in type 1. In order to test this hypothesis, the current study will attempt to clarify the precise mechanism of cell death that occurs in the diabetic state in order to identify targets for novel therapies. One of the ways this will be accomplished is by using transgenic mice that are prone to autoimmune diabetes as well as deficient for proteins that regulate cell death. In these mice, diabetes should develop more slowly, or not at all. This study will allow us to identify the pathways controlling beta cell damage from the immune system. Generating insulin-producing cells under conditions that prevent chemical stresses will provide better transplantation therapies for diabetes, and test the protection of beta cells from autoimmune killing in a human system to confirm mouse studies.
Techniques and Applications the Student will Learn: Mouse care, including food intake and body weight, glucose measurement, blood collection. protein assays, including A1c and insulin assays. Immunofluorescence analysis for islet size and cell viability Phosphoprotein blot analysis. Eligible Funding Sources: NIH Back to top Project Title: Pluripotent stem cells for diabetes
Mentor: Dr. Meri Firpo
Contact: firpo001@umn.edu
Co-investigator: N/A
Project Description: Our preliminary studies strongly support the conclusion that it is possible to cure diabetes with autologous cell transplantation. This strategy would allow us to transplant human islet cells without the requirement of anti-rejection medication. This project would be part of a graduate student and/or a postdocotral fellow, and would involve learning to perform cell culture for generating stem cells for differentiation, observing and participating in the differentiation under varied conditions to produce pancreatic and insulin-producing cells in culture from stem cells, and analyzing the reslting differentiation by expression analysis by quantitative PCR and immunofluorescence.
Techniques and Applications the Student will Learn: Participating Students in my laboratory will be provided with training in cell culture and molecular analysis techniques, as well as background training in stem cell and developmental biology and transplantation therapies. Participants will work on projects on human pancreatic development and transplantation therapies for type I diabetes. Eligible Funding Sources: NIH Back to top Project Title: Is Depression a Result of Faulty Thermoregulation?
Mentor: Dr. Alice A. Larson
Contact: larso011@umn.edu
Co-investigator: Dr. Katalin J. Kovacs
Project Description: Depression is a common condition that can occur spontaneously or result from stress. The etiology of depression is not fully understood. One animal model of depression may give us some clues. In this model, increases in floating/immobility by mice during a 5-minute forced swim in cold water are used as a sign of depressive behavior. Drugs that decrease this response often have antidepressant activity. We have found that the time spent immobile correlates highly with body temperature, whose regulation we understand much better than that of depression. If thermoregulation and depression are functionally linked, repeated changes in body temperature, brought about by daily warm or cold swims, might lead to a model of depression that is easy to produce and highly reproducible. Such a model might also be more easily assessed by changes in body temperature alone rather than the more time-consuming measurement of floating. We hypothesize that repeated hyperthermia or hypothermia leads to an adaptive thermoregulatory response as well as increased depressive behavior. To assess the relationship between thermoregulation and depression, we will monitor body temperature responses during and following exposure to warm and cold swims and determine whether temperature regulation is linked to the development of depressive behavior. Mice exposed to chronic stress or to manipulation of their brown adipose tissue are predicted to thermoregulate poorly and be more depressed than control mice. Techniques and Applications the Student will Learn: The student will learn to surgically implant telemetric devices that allow us to monitor thermoregulation in mice remotely; to surgically manipulate brown adipose and its sympathetic innervation; and to statistically analyze and graphically depict behavioral and telemetric data.
Eligible Funding Sources: NIH/CVM/Merial Back to top Project Title: Effect of Inflammation & Stress on Pain
Mentor: Dr. Alice A. Larson
Contact: larso011@umn.edu
Co-investigator: Dr. Katalin J. Kovacs
Project Description: Pain (referred to as nociception in animals) is increased by conditions such as inflammation or the surgical placement of a ligature on a nerve. In these conditions, the capsaicin-type receptor (TRPV1 receptor) is upregulated and plays a greater role in pain sensitivity (referred to as hyperalgesia). Stress also exacerbates musculoskeletal pain. This can be modeled in mice by the short-term increases in musculoskeletal hyperalgesia (measured by grip force responses) following a 15-min forced swim in cold water. Although swim stress-induced musculoskeletal hyperalgesia is mediated, in part, by CRF2 receptors, it does not involve TRPV1 receptors. We hypothesize that the intense musculoskeletal hyperalgesia observed in some conditions is due to the combination of stress with either inflammation or nerve ligation increasing pain more than either manipulation alone; this is likely due to the involvement of BOTH CRF2 receptors as well as TRPV1 receptors. We will test this possibility by subjecting mice to an inflammatory condition (Freund’s adjuvant injection) or to nerve ligation (surgical) and then test their musculoskeletal sensitivity to stress using the forced swim in cold water. We expect a synergistic hyperalgesia that is sensitive to morphine (verifying the effect on pain rather than strength), to CRF2 antagonists (verifying the involvement of stress hormones), and to TRPV1 receptor antagonists (verifying the role of capsaicin receptors). Techniques and Applications the Student will Learn: The student will learn surgical approaches to ligate spinal nerves in mice, chemical approaches to induce inflammation, behavioral measures of pain in rodents, statistical analyses of data, and graphical representation of data.
Eligible Funding Sources: NIH/CVM/Merial Back to top Project Title: Molecular Genetic Bases of Disease Traits in Dogs
Mentor: Dr. Jim Mickelson
Contact: micke001@umn.edu
Co-investigator: Dr. Ned Patterson
Project Description: Breeding practices which select for desirable traits in domestic animal species can inadvertently select for undesirable alleles that increase the prevalence of heritable diseases. A major goal of our canine genetic research group is to determine the genes and alleles that contribute to heritable neurological, neuromuscular and metabolic disorders in certain breeds. Our basic hypothesis is that as yet unidentified genes responsible for genetic traits can be identified by a "scan" of the genomes of dogs with a disorder (cases) and control dogs without the disorder with tens of thousands of DNA markers known as SNPs. Multiple comparisons of the allele frequencies of each of these SNP markers can then be made across case and control populations. This genome scan narrows the search for genes that contribute to that particular trait to relatively small chromosomal segments where the significantly different SNP markers are located, that contain relatively few plausible candidate genes. It is then possible to identify the causative mutation itself by sequencing DNA or mRNA from one or more genes in cases and comparing them to control horses. Any sequence differences (potential mutations) found are then analyzed in larger populations to accurately determine their association with the phenotype, and their frequency in the breed. Traits that could be pursued include peripheral neuropathies (Leonbergers and a number of breeds), urinary stone formation, urinary hypercalcimia, and diabetes mellitus (Miniature Schnauzers), idiopathic epilepsy (Australian Shepherds) and exercise-induced collapse (Border Collies).
Techniques and Applications the Student will Learn: The student will learn the principles behind the genome scanning methods, analyze SNP genotype data and determine allele frequencies across breeds, actively participate in the sequencing and mutation discovery processes that utilizes PCR, electrophoresis, and DNA sequence analysis software.
Eligible Funding Sources: NIH/CVM/Merial Back to top Project Title: Use of whole genome sequences to unravel the genetic basis of health, disease and performance in horses
Mentor: Dr. Molly McCue
Contact: mccu0173@umn.edu
Co-investigator: Dr. Jim Mickelson
Project Description: Breeding practices which select for desirable traits in domestic animal species can result in selection for and even fixation of the causative allele within breeds, as well as inadvertently select for undesirable alleles that increase the prevalence of heritable diseases. A major goal of our equine genetic research group is to determine the genes and alleles that contribute to musculoskeletal, neuromuscular and metabolic traits that have been highly selected for in horses, as well as genes and alleles that cause heritable disorders affecting these same systems in certain breeds. Our basic hypothesis is that as yet unidentified genes responsible for genetic traits can be identified by a "scan" of the genomes of hundreds of horses from dozens of breeds with tens of thousands of DNA markers known as SNPs. Multiple comparisons of the allele frequencies of each of these SNP markers can then be made across these populations (i.e., large vs small breeds; or disease cases vs healthy controls) to identify the regions of the genome harboring causative alleles. Although these approaches narrow the possible genomic regions down from the entire 2.7 gigabase equine genome, in many circumstances there are still several regions of the genome encompassing 2-10 megabases that need investigation. To simultaneously and efficiently investigate multiple regions of the genome, we are using next generation sequencing technologies to sequence the entire genome of 2-8 horses for each project. The goal of this summer scholars project is to analyze genomic sequences from horses to discover functional alleles (or mutations) responsible for traits of interest. Current projects using this approach include: Polysaccharide Storage Myopathy, Equine metabolic Syndrome, gait (pace/trot), racing performance, and performance traits in Quarter Horses.
Techniques and Applications the Student will Learn: The student will learn the principles behind the genome scanning and next generation sequencing, participate in the analysis of next-generation sequence data, and the discovery of novel variant/mutations in the horse. Student will participate in aligning relevant information from the genome to phenotypes of interest by genotyping of populations for discovered variants.
Eligible Funding Sources: NIH/CVM/Merial Back to top Project Title: Molecular mechanisms that drive osteosarcoma progression
Mentor: Dr. Jaime Modiano
Contact: modiano@umn.edu
Co-investigator: Dr. Subbaya Subramanian
Project Description: Osteosarcoma is a heterogeneous disease, but at present, treatment options are the same for every affected dog. Improving our understanding of what drives osteosarcoma progression will allow us to define better strategies and tailor treatments that will improve outcomes and reduce therapy-related adverse events. This project is designed to extend our recent findings that show canine and human osteosarcomas can be stratified into prognostically distinct molecular groups: one group is highly aggressive and another is more indolent. Here, we will define mechanisms that account for this stratification as the first step to develop interventions to cure or retard progression of highly aggressive osteosarcomas. The hypothesis is that inactivation of RB (a tumor suppressor protein) is causally related to the highly aggressive, rapidly progressing phenotype of canine and human osteosarcomas. The corollary is that restoring RB function (genetically or pharmacologically) will convert aggressive, rapidly progressing osteosarcomas to a more indolent phenotype, thus improving our ability to manage the most aggressive forms of this disease. The objective is to define differential protein expression induced by restoration of RB in canine osteosarcoma. We will use genetic approaches in a panel of canine and human osteosarcoma cell lines that retain “aggressive” (e.g., OSCA-40, OSCA-8, OSCA-78, Saos2) and “indolent” (e.g., OSCA-32, U2OS) gene expression phenotypes to establish the role of RB in defining patterns of protein expression. Restoration of RB will be achieved by introduction of wild type RB and genetically modified, "super-RB" using previously validated methods (nucleofection). Expression of RB will be verified by immunoblotting and immunoprecipitation. Differential protein expression will be examined in whole cell lysates, and differential complex formation will be examined from immunoprecipitated proteins. Association of RB onto specific gene promoters (silencing) will be examined using chromatin immunoprecipitation (ChIP) assays. Finally, we will evaluate acetylation and methylation tags in Histones to determine epigenetic changes in chromatin structure attributable to RB restoration. Special emphasis will be placed on genes that comprise prognostic osteosarcoma signatures. The summer scholar will present the project proposal at the beginning of summer at the Comparative Oncology Seminar; there will be regularly scheduled meetings and data presentation will be required at lab meetings and at the Annual Merial Summer Conference.
Techniques and Applications the Student will Learn: Laboratory safety, cell culture, recombinant DNA/cloning and plasmid expansion, transfection (nucleofection), quantitative real-time PCR, immunoblotting, flow cytometry, immunoprecipitation, chromatin immunoprecipitation, luciferase reporter assays, microscopy. The student also will learn data analysis and presentation skills.
Eligible Funding Sources: NIH/CVM/Merial/Oncology Fellow Back to top Project Title: CXCR7 signaling bias to transdifferentiate canine hemangiosarcoma
Mentor: Dr. Jaime Modiano
Contact: modiano@umn.edu
Co-investigator: Dr. Erin Dickerson
Project Description: Canine hemangiosarcoma is a common and lethal cancer, and the outcomes of dogs with this disease are unlikely to improve without a better understanding of its origin and pathogenesis. We have proposed that canine hemangiosarcoma arises from bone marrow progenitor cells. We now have multiple lines of evidence that support the hypotheses that tumor-initiating cells in hemangiosarcoma are multipotent, and that they are malleable to adapt to different microenvironments. For this project, we will examine the potential to use the multipotency of hemangiosarcoma cells to our advantage by forcing them to differentiate into lineages with reduced malignant potential. Hemangiosarcomas differ from normal endothelial cells by having greater propensity for inflammation and angiogenesis. More recent data show that hemangiosarcomas can be stratified according to the expression of pro-adipogenic and connective tissue-forming genes, endothelial cell-matrix interaction genes, and pro-inflammatory genes. These genes reflect the composition of the tumor microenvironment, and potentially the balance of hypoxia and inflammation during tumor formation. Differences in signaling by stromal-derived factor-1 (SDF-1) also could contribute to the observed heterogeneity of hemangiosarcoma. SDF-1 is produced both by tumor cells and by stromal cells. It binds two distinct chemokine receptors called CXCR4 and CXCR7. The role of CXCR7 in particular is not fully understood: it may contribute to malignancy in some tumors; in normal cells, it is required for hematopoietic development and for fat and bone formation; and CXCR7 signals may instruct tumor cells to secrete factors that recruit inflammatory and angiogenic cells into the tumor environment. We will examine if signaling bias through CXCR7 can be used to enhance adipogenic differentiation of hemangiosarcoma cells, and conversely if CXCR7 blockade can promote inflammatory and angiogenic phenotypes (by enhancing CXCR4 signaling). The objectives will be to assess differentiation of hemangiosarcoma cells in culture under conditions of logarithmic growth with and without SDF-1. We will use targeted genetic mutants and small molecule inhibitors to bias SDF-1 signals towards CXCR7 or CXCR4, and will assess self-renewal of hemangiosarcoma cells, differentiation towards fat-formation, and differentiation towards inflammatory or vascular cell formation in culture. This supervised project will include use of novel drugs obtained through a collaboration with an industry partner. The summer scholar will present the project proposal at the beginning of summer at the Comparative Oncology Seminar; there will be regularly scheduled meetings and data presentation will be required at lab meetings and at the Annual Merial Summer Conference.
Techniques and Applications the Student will Learn: Laboratory safety, cell culture, recombinant DNA/cloning and plasmid expansion, transfection, non-adherent sphere forming efficiency to quantify self-renewal, quantitative real-time PCR and flow cytometry to determine expression of vascular, adipogenic, and hematopoietic differentiation markers, formation of vessel networks in Matrigel, phagocytic activity, and CFSE dilution assays to assess proliferation. The student also will learn data analysis and presentation skills.
Eligible Funding Sources: NIH/CVM/Merial/Oncology Fellow Back to top Project Title: Understanding and enhancing mechanisms of priming in cancer immunotherapy
Mentor: Dr. Liz Pluhar
Contact: pluha006@umn.edu
Co-investigator: Dr. John Ohlfest
Project Description: 1) Background and Significance: Gliomas are one of the most common primary brain tumor in dogs with a median survival of 3 months despite treatment with currently used methods. We are trying to develop a new treatment method to increase the survival time in client-owned dogs that have spontaneously occurring gliomas. 2) Question being addressed: Does treatment with gene therapy or vaccines created from the dog’s own tumor kill residual tumor cells after surgery and prevent regrowth of tumor by stimulating the dog’s immune system. 3) How will the results of the study be used?: If successful, as a new treatment option for other dogs with gliomas, and as a model to develop a similar treatment protocol for people with gliomas. 4) Summarize the Specific Aims: To show that dogs treated by this method create antibodies against the tumor cells that are capable of indirect killing of tumor cells and immune cells that can directly kill tumor cells. To demonstrate that this treatment method is safe and increases survival.
Techniques and Applications the Student will Learn: There are many aspects of the study in which the student can be involved depending on their interest: reading serial MRI scans and measuring tumor volumes, interpretation of blood work in dogs treated in the study, tumor cell culture and vaccine production, gel electrophoresis and Western blots for immune monitoring.
Eligible Funding Sources: NIH/CVM/Merial/Oncology Fellow Back to top Project Title: FT-IR Microscopy in Osteoporosis
Mentor: Dr. Liz Pluhar
Contact: pluha006@umn.edu
Co-investigator: Dr. John Ohlfest
Project Description: 1) Background and Significance (1-2 sentences): Currently measurement of bone mineral density (BMD) is the most widely used method to assess osteoporosis and predict fracture. However, it turns out that BMD is not highly associated with the incidence of fracture in people with osteoporosis and osteopenia. Earlier studies with FTIRI of bone biopsies showed that this method accurately reflects specific mineral and material properties of bone. 2) Question being addressed (1-2 sentences): The goal of this project is to assess how well a new method of measuring bone mineral density, called Fourier Transform infrared spectroscopic (FTIR) imaging (FTIRI), predicts osteoporosis and the risk of bone fracture. We hope to show that FTIRI parameters correlate with the propensity to develop fractures and can be used as a better predictive tool in osteoporotic patients. 3) How will the results of the study be used? (1-2 sentences): Fourier Transform infrared spectroscopic (FTIR) imaging (FTIRI), will be used to predict osteoporosis and the risk of bone fracture in women and their response to osteoporosis treatments. 4) Summarize the Specific Aims (derived from the grant proposal): We propose to use an animal model of severe osteoporosis, adult female sheep with their ovaries removed that have been placed on a special diet that causes them to lose mineral from their bones, to test the ability of FTIRI to measure the material properties of bone and to predict bone that will fracture. We will test this in sheep with osteoporosis and sheep with osteoporosis that are being treated with an agent that may reverse osteoporosis called parathyroid hormone.
Techniques and Applications the Student will Learn: Data acquisition and interpretation of DEXA scans including bone mineral density (BMD) and bone mineral content (BMC) and how osteoporosis and PTH affect BMD and BMC. Laboratory techniques to process serum and urine samples and perform ELISA on samples to measure markers of bone metabolism and interpret the effects of osteoporosis and PTH therapy on these markers.
Eligible Funding Sources: NIH/CVM/Merial Back to top Project Title: Mechanisms of inflammation regulation
Mentor: Dr. Bruce Walcheck
Contact: walch003@umn.edu
Co-investigator: N/A
Project Description: My lab is examining various aspects of inflammation regulation. We are interested in mechanisms that direct white blood cells (leukocytes) to sites of inflammation and that regulate their activities. A key leukocyte involved in the early inflammatory response is the neutrophil, which is the most abundant leukocyte in the blood and at sites of acute inflammation. Various factors that regulate inflammation are released from the surface of neutrophils by a process referred to as ectodomain shedding, including the very potent cytokine TNF-alpha. My lab is actively involved in understanding the function and regulation of this proteolytic process in order to enhance the immune response or reduce damaging inflammation.
Techniques and Applications the Student will Learn: Leukocyte isolation from animal and human blood. Antibody-based assays such as flow cytometry, Western blot, and ElISA Molecular biology Tissue culture techniques perhaps animal handling, such as mice
Eligible Funding Sources: NIH/CVM/Merial
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Project Title: Utilization of protein splicing to generate highly functional dystrophin constructs for gene therapy applications.
Mentor: Dr. DeWayne Townsend
Contact: town0045@umn.edu
Co-Investigator: N/A Project Description: Duchenne muscular dystrophy is a devastating uniformly fatal neuromuscular disease resulting from the loss of the protein dystrophin. Dystrophin's large size (11,000 bp) precludes it direct replacement by gene therapy vectors, whose carrying capacity is 4500 bp. Detailed structure function studies have provided information on critical structural domains allowing the formation of highly truncated dystrophin constructs that can be carried in gene therapy vectors. Not surprisingly, these truncated dystrophins do not completely replace the full-length dystrophin. Recent studies have focused on finding new methods to deliver larger dystrophin constructs, however most of these processes are inefficient resulting in sub-optimal dystrophin expression. Protein splicing utilizes unique bacterial proteins that function to fuse two poly-peptides. This approach promises to allow delivery of large dystrophins with great efficiency, a critical feature of any viable gene therapy approach. Techniques and Applications the Student will Learn: Polymerase chain reaction Standard cloning techniques Protein Purification SDS-PAGE Western Blotting
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Emerging Infectious Diseases & Zoonoses
Project Title: Characterizing T cell responses to swine influenza virus
Mentor: Dr. Maxim Cheeran
Contact: cheeran@umn.edu
Co-investigators: Dr. Montse Torremorell, Dr. Srinand Sreevatsan, Dr. Thomas Molitor Project Description: Influenza viruses are well known for their ability to generate antigenic variations that consequently enable immune evasion. This viral characteristic is the bane of vaccine efforts against influenza, including in domestic swine production. Antibody response is the predominant correlate for protection during influenza infections and has been extensively studied for its effect on generating strain variations. While it is clear that the CMI responses play a significant role in cross protection against influenza virus subtypes, there is a significant gap in knowledge regarding its influence on antigenic variation. Our laboratory studies cell-mediated immune responses to SIV with the goal: To identify the major SIV determinants of porcine immune response and to determine how these determinants change on the virus under selective immune pressure. Initial efforts in the lab are focused on understanding the SIV epitopes recognized by porcine T lymphocytes in response to infection and/or vaccination. These studies are expected to provide tools required to assess T cell responses against emerging influenza viruses in the domestic swine population that would help in predicting protection and developing novel vaccine strategies.
Techniques and Applications the Student will Learn: Techniques the student will learn and use: Lymphocyte isolation from Porcine blood and bronchoalveolar lavage fluid, ELISPOT assay for T cell function, Flowcytometry, lymphocyte separation and RT-PCR for porcine cytokines.
Eligible Funding Sources: SDEC Back to top Project Title: Characterizing mucosal immune responses to swine respiratory pathogens
Mentor: Dr. Thomas Molitor or Dr. Maxim Cheeran
Contact: molit001@umn.edu, cheeran@umn.edu
Co-investigators: Dr. Maria Pieters, Dr. Montse Toremorell, Dr. Albert Rovira Project Description: Respiratory infections with pathogens, such as swine influenza virus, Hemophillus parasuis, and Mycoplasma hyopneumoniae, are characterized are by robust mucosal responses mediated by innate and adaptive immune mechanisms. While these infections result in substantial economic losses to the swine industry, commercial vaccines available offer less than complete protection against infection albeit they protect against severe disease. This vaccination strategy fosters an environment for evolution of pathogens that evade the host response, potentially resulting in emergence of novel strains. Majority of vaccine efficacy studies utilize humoral immune correlates to determine level of protection, with little to no data available on responses elicited at the mucosa, the primary site of infection. This gap in knowledge is particularly evident in understanding the role of local T cell responses in conferring protection against these pathogens. The present summer project will evaluate the humoral and cell-mediated immune responses in the lung and bronchial lymph nodes to swine Influenza virus, M. hyopneumoniae, or H parasuis. The primary objectives are to determine if vaccination: (1) alters humoral response patterns to pathogen challenge, including IgA and cytokine response, and (2) accentuates local antigen specific T cell responses in the lung and draining lymph nodes. Ultimately, identifying mucosal responses that confer protection would help develop vaccine strategies that not only provide protective immunity at the respiratory mucosa but also protect against transmission within the swine population. Techniques and Applications the Student will Learn: Lymphocyte isolation from Porcine blood and bronchoalveolar lavage fluid, ELISPOT assay for T and B cell function, lymphocyte separation and RT-PCR and ELISA for porcine cytokines. Eligible Funding Sources: VPM/SDEC Back to top Project Title: Interaction of anti-circovirus immunity in sows and anti-circovirus immune responses in piglets
Mentor: Dr. Michael Murtaugh
Contact: murta001@umn.edu
Co-investigator: Dr. Cheryl Dvorak Project Description: Porcine circovirus 2 (PCV2) is the causative agent of PCV-associated disease (PCVAD), one of the most economically important diseases of pigs. Widespread vaccination of pigs starting in 2006 has ameliorated PCVAD in finishers so successfully that nearly all pigs in the US are currently vaccinated at or before weaning. However, sow infection and in utero transmission suggest a reproductive effect of PCV2 infection. In fact, vaccination of sows before farrowing in addition to piglet vaccination at weaning has been shown to decrease post-weaning mortality, decrease viremia, and increase production performance. However, other studies suggest that maternal antibodies interfere with the efficacy of piglet vaccination. The goal of this project is to determine if PCV2 vaccination of sows pre-farrow affects the efficacy of protection by PCV2 vaccine when given to piglets at 3 weeks of age. Sows will be vaccinated pre-farrow and piglets will be examined at weaning and during finishing for PCV2-specific antibody levels and PCV2 viral loads in serum. Students will travel to a pig farm in southern MN to help vaccinate sows and later to tag and sample piglets at weaning and during finishing from vaccinated and non-vaccinated sows. Serum samples from weaned and finishing pigs will be examined for PCV2-specific antibodies using ELISA and serum neutralization assays and examined for PCV2 viral loads using qPCR.
Techniques and Applications the Student will Learn: Students will be introduced to basic immunological procedures used for analysis of biochemical and functional immune response, including ELISA and viral neutralization. They will learn about experimental design, the purpose of controls, importance of sample size, basic statistical analysis of experimental data, and the value of collaborative group work. Eligible Funding Sources: SDEC Back to top Project Title: Characterization of macrophage permissivity to porcine reproductive and respiratory syndrome virus
Mentor: Dr. Michael Murtaugh
Contact: murta001@umn.edu
Co-investigator: Sally Robinson Project Description: Molecular requirements for cell infection by porcine reproductive and respiratory syndrome virus (PRRSV) are a central, unanswered question complicating the development of more effective immunological control measures for prevention of PRRS. We have developed quantitative methods to assess macrophage infection by PRRSV that will be used to examine the relationship between age of pigs, CD163 expression, and permissivity. We have developed preliminary data showing that macrophages from young pigs are more permissive than adult macrophages to PRRSV infection. Patton et al. (2009) showed that treating CD14+ blood monocytes with IL-10 during differentiation into macrophages increased expression of CD163 and correspondingly increased PRRSV infection. We expect to compare young versus old pig macrophages for differences in CD163 expression, and possibly for variation in IL-10 responsiveness and gene expression associated with permissiveness to PRRSV infection. Techniques and Applications the Student will Learn: The student will be introduced to basic cell biology and virological techniques used to study host-pathogen interactions, including sterile eukaryotic cell culture methods, FACS, viral growth and growth analysis, determinations of viral titer and ELISA. They will learn about experimental design, the purpose of controls, importance of sample size, basic statistical analysis of experimental data, and the meaning of randomness. Eligible Funding Sources: SDEC Back to top Project Title: Location, abundance, and proliferation status of SIV-specific CD8 T cells in situ in rhesus macaques vaccinated with live-attenuated SIV vaccines and challenged with pathogenic SIV
Mentor: Dr. Pamela Skinner
Contact: skinn002@umn.edu
Co-investigator: N/A Project Description: Live-attenuated SIV vaccines (LAVs) provide the best protection of all HIV/SIV vaccines to date, yet the mechanisms of protection are not completely understood. To gain insights into mechanisms of protection by LAVs and aid the development of effective HIV vaccines, we study the localization, abundance and proliferation status of SIV-specific CD8 T cells in situ in genital and lymphoid tissues from rhesus macaques after vaccination with LAVs and after intravaginal challenge with pathogenic SIV. Techniques and Applications the Student will Learn: In situ tetramer staining, confocal microscopy, and quantitative image analysis. Eligible Funding Sources: NIH/CVM/Merial Back to top Project Title: Serological Evidence of pathogenic Paramyxoviruses circulating in wild birds in the Upper Midwest
Mentor: Dr. Patrick T. Redig
Contact: redig001@umn.edu
Co-investigator: Dr. Carol Cardona Project Description: Since the early 1990’s, pathogenic strains of avian paramyxovirus (pPMV and cPMV), variants of Newcastle disease virus (PMV-1) found ion pigeons and cormorants, respectively, have been present in the prairie regions of North America. In recent years, a pattern of every-other-year epizootics, especially in cormorants, have been recorded in Minnesota. These outbreaks have been occurring in the region of shallow lakes in tall grass prairie regions of the state, extending as far north as Leech Lake. This is a region of intense poultry production in the state and there is the potential for spill-over of these viruses, which have pathogenic potential for chickens and turkeys. The factors driving this episodic nature of outbreaks are unknown; however, it is hypothesized that it may be due to a waxing and waning of herd immunity. In 2012, it was demonstrated that a competitive ELISA test was effective in identifying antibodies to paramyxoviruses in the yolk of eggs laid by cormorants; a 5% prevalence was recorded from a sample of 30 eggs. We propose to use this test to determine antibody prevalence in cormorant eggs in a long-term, year-over-year study to correlate fluctuations in antibody prevalence in eggs with the episodic outbreaks.
Techniques and Applications the Student will Learn: 1. Procedures for conducting ELISA-based analysis of egg yolk material 2. Data keeping and analysis 3. Interpretation and presentation of data 4. Additionally, a summer scholar working on this project may spend 15 – 20 hours a week working in The Raptor Center Clinic obtaining experience in clinical procedures on raptors Eligible Funding Sources: Raptor Center Back to top Project Title: Evaluation of bacteriophages to inactivate antibiotic-resistant shiga toxigenic E. coli non-O157:H7 strains on food matrices
Mentor: Dr. Fernando Sampedro
Contact: fsampedr@umn.edu
Co-investigator: Dr. Mastura Akthar Project Description: Background: In the United States, non-O157 Escherichia coli may account for 20-50% of all shiga toxigenic E. coli (STEC) infections, as well as associated with foodborne outbreaks. Cattle serve as reservoir for numerous STEC. The emergence of antibiotic resistant STEC has been already linked with raw meats which might pose a higher risk to human health. Different strategies have been adopted to control these foodborne bacteria in food processing plants. This study will evaluate bacteriophages to reduce resistant non-O157 STEC from food matrices as an alternate biological control approach. Hypothesis: Bacteriophages may reduce the STEC contamination on different food matrices by lysing the target pathogens. Objective: The objective of this study was to evaluate the effects of E. coli specific bacteriophages on the viability of antibiotic resistant non-O157 STEC strains on surfaces of food representative of food processing plants. Methods: In this study, E. coli specific bacteriophages will be used that were previously isolated (Stelios et. al. 2011) in Food Safety Microbiology Lab. Efficacy of phage treatments will be determined on food sample (raw meat, RTE meat) surfaces. All food samples will be inoculated with bacteria (103-104 CFU/mL) at 23˚C, and then treated with phages. The degree of bacterial lysis will be determined at different times (0, 30 min, 2 hours, and 24 hours) temperatures (4, 10, 23° C) and plated on respective antibiotic plates. Bacterial survivors will be identified with and without phage treatment and the inactivation will be calculated. These study findings will suggest phages can serve as an effective and natural control strategy to reduce the incidence of foodborne antibiotic resistant non-O157 STEC. Techniques and Applications the Student will Learn: Microbiology techniques: Bacteriophage culturing, Data analysis Eligible Funding Sources: VPM Back to top Project Title: Simulation modeling of foodborne pathogen transmission, sampling strategies, and antimicrobial resistance (multiple projects)
Mentor: Dr. Randall Singer
Contact: rsinger@umn.edu
Co-investigator: N/A Project Description: In this project, you will learn to develop simulation models using programs such as Microsoft Excel or modeling software such as STELLA. There are several questions that can be addressed in these models. In one type of project, the student can assess the optimal sampling strategy for detecting the diversity of antimicrobial resistance patterns that exist on a farm or in a retail meat sample. This type of model will aid in the development of on-farm and retail surveillance systems such as NARMS (I collaborate with FDA and USDA). In another project, the student can choose to develop simulation models predicting the spread of antimicrobial resistance on the farm. Finally, the student can help build models of the poultry production system that will aid in the prediction of Campylobacter or Salmonella being transmitted to humans through the consumption of poultry products. If the student has ideas for a completely different type of simulation model, I am open to additional suggestions for projects. Techniques and Applications the Student will Learn: The student will learn to formalize equations of disease transmission into software such as Microsoft Excel with @RISK or use systems modeling software such as STELLA. With these programs, the student will gain a deeper understanding of risk (risk assessment), as the models will likely be related to human health risks associated with certain activities or production systems. During the project, the student will learn to take data from the published literature and use it to estimate parameters of transmission. Finally, the student will learn how to prepare a scientific paper in the area of simulation modeling / risk assessment. Eligible Funding Sources: TBD Back to top Project Title: Characterization of bacterial isolates from Daurian pikas with a systemic bacterial infection
Mentor: Dr. Arno Wuenschmann
Contact: wunsc001@umn.edu
Co-investigator: Dr. Connie Gebhart Project Description: Four Daurian pikas from a zoo collection submitted for necropsy between October 2012 and January 2013 had lesions consistent with a bacterial infection targeting lymphoid organs and other internal organs. Gram-negative, non-fermenting bacteria were isolated from each case but further classification and identification of the bacteria was not possible using routine methods. The project aims at 1) describing the lesions and 2) further characterizing the bacterial isolates. Techniques and Applications the Student will Learn: 1) Review cases including histopathology and staining characteristics of the bacteria in histological sections. 2) Further characterization and possible speciation of isolates (are the isolates identical? which genus/species are they?). A) Thawing and re-culturing of frozen isolates B) Biochemical analysis C) Sensitivity testing D) 16S gene sequencing E) Appling MADLI technology The project could be extended if needed to include the following: 3) Collect fecal samples from live animals and environmental samples for bacteriology in an attempt to find out whether the isolate is being shed via fecal route and to check whether the isolate is present in the environment. 4) Collect more samples at necropsy if additional animals die of this condition in the future. 5) Explore whether vaccine are feasible. Eligible Funding Sources: VPM Back to top
Population Medicine Project Title: Domestic dogs as sentinels for anthrax exposure
Mentor: Dr. Meggan Craft
Contact: craft@umn.edu
Co-investigator: N/A Project Description: Anthrax, an ancient disease, is a disease of global importance, affecting humans, livestock and wildlife. In nature, anthrax is present in the soil and has a broad geographic range, but the conditions under which it is maintained are still poorly understood. Carnivores and humans typically get infected through consumption of, or contact with, infected herbivore carcasses. It has recently been shown that domestic dogs serve as good indicators, or sentinels, of human and livestock anthrax risk. Domestic dogs can act as bioaccumulators of infection through ingestion of infected carcasses, are highly resistant to anthrax, and can have antibodies in their blood that indicate if anthrax has been present in the animal populations in their local environment during any period of their life. Anthrax provides a seminal case study to investigate where land-based livelihoods, lack of animal and public health infrastructure, extreme poverty, and environmental pathogens intersect. This study area, Queen Elizabeth National Park in Uganda, is a region with known anthrax outbreaks. The main goal of this Grant-in-Aid is to conduct a pilot study to identify social and environmental factors that predict spatial and temporal patterns of anthrax exposure in a key sentinel species, domestic dogs, and thus of lethal disease to associated humans, livestock and wildlife. Two steps are required for this project, (i) development of a domestic dog anthrax serology test, and (ii) collection of dog sera and questionnaire data. Eligible Funding Sources: VPM
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Project Title: Ecology of Eastern Equine Encepalatis in mosquitoes and wildlife in Minnesota
Mentor: Dr. Meggan Craft
Contact: craft@umn.edu
Co-investigator: N/A
Project Description: A research opportunity will be available to build upon a successful 2012 Summer Scholars project to better understand the ecology of eastern equine encephalitis virus (EEEv) in wildlife and mosquitoes in Minnesota. EEEv is a mosquito-borne zoonotic virus. The primary transmission cycle involves mosquitoes and wild birds as reservoir hosts; infection has been documented in horses, humans and wildlife species. The aim of the previous study was to examine vectors of EEEV in relation to high antibody titers found in moose and elk that were sampled as part of the Minnesota Department of Natural Resources’ wildlife health surveillance efforts. Depending on which mosquitoes (if any) test positive for EEEv, future directions for this project might include: sampling mosquitoes with multiple types of traps and determining which wild bird or wild animal host mosquitoes have fed on through blood meal analysis. This project is a collaboration between UMN and the Minnesota Department of Health, the Minnesota Department of Natural Resources, and the Metropolitan Mosquito Control District. Techniques and Applications the Student will Learn: Working in collaborative groups, independent research, organizational skills, personnel management, mosquito identification, study design, basic statistical techniques and others. Eligible Funding Sources: VPM
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Project Title: Epidemiology of Staphylococcus aureus in pigs and swine veterinarians
Mentor: Dr. Peter Davies
Contact: davie001@umn.edu
Co-investigator: N/A
Project Description: Certain recently identified strains of S. aureus that are adapted to livestock (including pigs) have raised concerns about food animals as reservoirs of MRSA infection of people. We have ongoing studies looking at the epidemiology of S. aureus in pigs and the risk of tranmission to humans (specifically S. aureus). The student will participate in laboratory and field components of ongoing studies of S. aureus in pigs and swine veterinarians, focusing on livestock associated MRSA. Much of the work will be involved with processing of regular samples being submitted to the lab, and will involve selective and non-selective culture, basis tests for confirmation and identification of S. aureus (catalase, coagulase, S. aureus agglutination tests). The student will also learn PCR and sequencing methods used for subtyping and characterization of the isolates. A field component involving more pig studies is also proposed and funding is pending. It is hoped that the student will pursue a smaller project focused on antimicrobial resistance for presentation at the 2014 AASV meeting Techniques and Applications the Student will Learn: Routine bacteriology - culture and identification of S. aureus PCR and sub-typing methods - spa typing, MLST, mecA gene Antimicrobial resistance testing Eligible Funding Sources: SDEC
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Project Title: Clinical impact of transfaunation in the treatment of post-operative dairy cows
Mentor: Dr. John Fetrow
Contact: fetrow@umn.edu
Co-investigators: Dr. Erin Royster, Dr. Sandra Godden, Dr. Ricardo Chebel
Project Description: Study description Treatment of cattle with left-sided displacement of the abomasum (LDA) involves surgical repair of the malposition and correction of metabolic disturbances. Cattle with LDA respond favorably to surgical treatment; however, substantial economic losses as a result of delayed or decreased peak milk production, metabolic disturbances, and increased culling rates are encountered. 2,3 Ancillary medical treatments for LDA include administration of electrolyte solutions, calcium salts, sodium propionate, and propylene glycol. Metabolic disorders in inappetant cows include abnormal rumen fermentation, low energy intake, ketonemia, low blood and rumen concentrations of volatile fatty acids (VFAs), increased fat mobilization, and high serum concentrations of non-esterified fatty acids (NEFAs). Many of these metabolic disturbances either reflect inadequate rumen fermentation of reduction of feed intake and inadequate nutrient delivery to the cow. Rumen fermentation problems reflect changes in types or numbers of rumen microflora due to the cow eating less or due to electrolyte changes in the rumen contents. After surgery, the cow may benefit from re-inoculation of healthy rumen contents to regenerate a normal flora. A small scale study under teaching hospital conditions showed some metabolic improvements to such therapy, but the techniques has not been studied under commercial dairy conditions in a large sample of cows and the long term outcome of survival and reproductive and productive performance was not addressed. This study will involve transfaunation of a randomly selected population of cows post-surgery at the college's Dairy Education Center during the summer of 2013. We estimate there will be approximately 100 cases during that period, providing a much larger sample for treatment (transfaunation) and control (water) groups. Cows will be followed after treatment for metabolic status (urine ketones, blood glucose), milk production (daily for two weeks, the monthly and for the full lactation) and longer term production, reproductive performance, and survival in the herd. The student will live during the week at the dairy in the student dormitories to be on site for case enrollment and sample and data capture. Techniques and Applications the Student will Learn: Clinical trial design, including case definition, subject enrollment, statistical power clinical trial conduct data capture and data management data analysis: descriptive statistics, statistical testing: ANOVA/regression, survival analysis, and logistic regression In addition, the student will be immersed in the large commercial dairy with an active veterinary teaching program and continuing education program and will have an opportunity to learn a great deal about dairy management and veterinary medicine during the summer. Eligible Funding Sources: VPM
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Project Title: Supplementing Poor Quality Colostrum with a Colostrum Replacer: Effects on Passive Transfer of IgG and Calf Health
Mentor: Dr. Sandra Godden
Contact: godde002@umn.edu
Co-investigator: N/A
Project Description: Successful passive transfer of colostral immunoglobulins (e.g. IgG) is a critical determinant of calf health. In order to attain high serum IgG concentrations in the calf, is recommended that calves receive between 150 to 200 g of IgG in the first colostrum feeding, fed within 1 to 2 hours of birth. Unfortunately colostrum quality is highly variable among cows. A recent study of four U.S. dairy regions, reported that 30% of 827 colostrum samples tested were of poor quality (< 50 g/L IgG). Calves that are fed poor quality colostrum are at increased risk of failure of passive transfer and related health problems. One possible solution to this problem may be to use a Brix refractometer as a cow-side test to estimate the concentration of IgG in colostrum, and then supplement poorer quality colostrum with a powdered colostrum replacer product to achieve a target intake of 200 g of IgG in the mixture fed to the calf. Research needs to be conducted to measure whether this approach can be used to successfully achieve high levels of passive transfer (serum IgG) in the calf. At the same time, we must study whether this approach could have negative consequences such as decreased efficiency of IgG absorption, or creating a hyperosmotic solution that could produce diarrhea in the calf. This study will be conducted on a large dairy farm. Objectives will be to evaluate the effect of supplementing poor quality colostrum with a colostrum replacer on the following outcomes: i) total mass (g) of IgG ingested, ii) IgG concentration and osmolality of the colostrum/replacer mixture fed, iii) efficiency of absorption of IgG (%) in the calf, iv) serum IgG (mg/ml) in the calf at 24 hours of age, and v) health scores, including fecal consistency scores, in the calf. Our hypothesis is that calves can be safely fed colostrum supplemented with a colostrum replacer powder, resulting in high serum IgG concentrations in the calf, while having no negative effects on efficiency of IgG absorption or calf health. Techniques and Applications the Student will Learn: In addition to gaining experience related to study activities working with calves (jugular venipuncture, esophageal tube feeder, bottle feeding), the student will have an opportunity to assist farm staff with other daily management activities including obstetrical procedures, examination and treatment of sick animals, and assisting with the occasional DA surgery. The student will have an opportunity to present their results at the Annual Meeting of the American Association of Bovine Practitioners (AABP) in fall, 2014. Eligible Funding Sources: VPM
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Project Title: Tissue transcriptome analysis in the horse
Mentor: Dr. Molly McCue
Contact: mccu0173@umn.edu
Co-investigator: Dr. Jim Mickelson Project Description: A major goal of our equine genetic research group is to determine the genes and alleles that contribute to musculoskeletal, neuromuscular and metabolic traits in horses including the study of several important diseases such as Polysaccharide Storage Myopathy, Equine Metabolic Syndrome and Osteochondrosis. For all of these projects we are using whole genome association to ‘scan’ to genome and identify regions that likely contain the gene(s) that contribute to that particular trait. Once a genomic region is identified, plausible candidate genes are identified for sequencing, based on know function of the gene and tissue expression patterns. However, in the horse, relatively little information is known about the gene(s) and structure (alternate splicing, etc) in tissues; oftentimes making it difficult to identify relevant candidate genes and sequence these genes one identified. To overcome this problem our group has initiated a large scale study of the transcriptome (all the mRNA transcripts in the cell) of several tissues in the horse. The transcriptome reflects the genes that are being actively expressed at any given time, as well as the structure of the mRNA in the tissue. To capture the transcriptome of a tissue, we have used RNA-sequencing (RNA-seq, a ‘next-generation” sequencing technology) that allows for simultaneous sequencing of all mRNA present in the tissue at the time of sampling. The goal of this summer scholar project is to use RNA-seq data to reconstruct the transcriptome of one or more revenant equine tissues such as lamina, muscle, adipose, liver, cartilage, etc. This will provide novel insight into patterns of gene expression and structure in certain tissues; this data will then be used to select biologic candidate genes for diseases related to this tissue (eg Osteochondrosis and cartilage). Techniques and Applications the Student will Learn: The student will learn the principles behind the genome scanning and next generation sequencing, participate in the analysis of next-generation sequence data, and the discovery of novel sequences and splice variants in equine tissues. Student will participate in aligning relevant information from the transcriptome to discoveries form genome scans to identify candidate genes. Eligible Funding Sources: NIH/CVM/Merial
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Project Title: Tick-borne dsease ecology in Minnesota
Mentor: Dr. Katey Pelican
Contact: pelicank@umn.edu
Co-investigator: Dr. Meggan Craft Project Description: Lyme disease emerged in Minnesota in the 1980s and has spread in a distinct spatial pattern over the past several decades. Currently, Lyme Disease is endemic in the eastern regions of the state and still expanding in the westerly direction. At the same time, new tick-borne diseases have been identified in the state in recent years that are of concern to human health. Rodents are key host species for Lyme disease and other tick-borne disease in the state. A better understanding of tick borne disease ecology in rodents is a key step in understanding the ecology of tick-borne disease in the state. This project will link Lyme disease research in the state to vertebrate research in the UMN run long-term ecological monitoring site, Cedar Creek Ecosystem Science Reserve. Techniques and Applications the Student will Learn: Techniques the student will learn and use: 1. GIS modeling and analysis 2. Tick and host collection and speciation 3. Rodent host collection, speciation and sampling 4. Literature review and analysis Eligible Funding Sources: VPM
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Project Title: Developing a tool for assessing immunotoxicology in bald eagles
Mentor: Dr. Julia Ponder
Contact: ponde003@umn.edu
Co-investigator: Dr. Patrick Redig, Dr. Michelle Willette
Project Description: Wildlife is increasingly exposed to a broad array of environmental contaminants. As apex predators, raptors are sentinels for ecosystem health. While little is known about the sub-lethal effects of most of these contaminants, immune function is recognized as a sensitive indicator of contaminant exposure. We propose to use bald eagles to validate immune function analytic techniques and develop a method for screening for immunotoxicity using lead as a model for an environmental contaminant. The specific objective of this study is to conduct and validate immunological assessments in several species of captive raptors. These raptors are permanently injured and captivity-acclimated birds in The Raptor Center's (TRC) education program and will be used as a control group. This study will establish preliminary data against which the effects of environmental contaminants such as lead may be assessed in future studies. Immune function parameters could include: complete blood cell counts and complementary hematology, fibrinogen levels, humoral response to injected sRBC's and T-cell response to injected phytohemagglutinin (PHA). Depending on funding availablility, acute phase proteins and plasma protein electrophoresis could be included. Feces will also be collected, analyzed for ova and parasites and processed for future fecal glucocorticoid testing. Techniques and Applications the Student will Learn: Avian handling and restraint; sample collection (handling, processing and curation); clinical pathology techniques; data collection and analysis; organizing and summarizing data and findings; scientific literature search, scientific writing and communication and dissemination of scientific information. Eligible Funding Sources: Merial/CVM/Raptor Center
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Project Title: Establish normal coagulation reference ranges in several species of raptors
Mentor: Dr. Julia Ponder
Contact: ponde003@umn.edu
Co-investigator: Dr. Patrick Redig, Dr. Michelle Willette
Project Description: This research will build on The Raptor Center’s (TRC) recent work on a rodent eradication project using brodifacoum in the Galapagos Islands. We are developing a sustained-release delivery system for vitamin K to be used in the mitigation of secondary poisoning of vulnerable hawk populations. Prior to determining the efficacy of the vitamin K implant, normal coagulation times must be established for these species. Standard methods of assessing coagulation in birds include prothrombin time (PT), activated clotting time (ACT), and Russell’s viper venom time (RVVT). Both PT and RVVT analysis require clinical laboratory facilities, whereas ACT analysis is considered a point-of-care assay and can be used as a screening test. This research will also have a practical application for clinical wildlife medicine as well as avian and zoological medicine. In a recent study, “virtually 100% of the owls and a large percentage of hawks had post-mortem residues of at least one anticoagulant rodenticide in their livers.” Coagulation time can be used as a metric for diagnosing rodenticide toxicity and monitoring treatment. Establishing ACT reference ranges as a point-of-care screening test will give wildlife rehabilitators an additional diagnostic tool for raptors admitted to rehabilitation hospitals. The specific objective of this study is to establish normal coagulation reference ranges in several species of raptors. Scholar will have access to both captive (education) and wild raptors admitted for rehabilitation. All blood samples will be obtained during routine preventive or clinical medical procedures. Depending upon funding availability, coagulation results may be correlated to liver anticoagulant rodenticide levels for those wild raptors which die or are euthanized for humane reasons. Techniques and Applications the Student will Learn: Avian handling and restraint; sample collection (handling, processing and curation); clinical pathology techniques; data collection and analysis; organizing and summarizing data and findings; scientific literature search, scientific writing and communication and dissemination of scientific information. Eligible Funding Sources: Merial/CVM/Raptor Center
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Project Title: Are horses more lame at a walk then we think? A force plate analysis of equine gait.
Mentor: Dr. Troy Trumble
Contact: trumb016@umn.edu
Co-investigator: N/A
Project Description: Lameness has long been the most common costly disease to affect the horse industry. In order to minimize losses due to lameness, equine practitioners must be able to diagnose lameness accurately and efficiently, so they can provide targeted treatment. One obstacle practitioner’s face is the inherent subjectivity in the most common method of lameness diagnosis. In a clinical setting, lameness diagnosticians rely primarily on visual clues indicative of gait asymmetry to detect lameness. Previous horse research has combined traditional subjective analysis with force plate data to describe lameness, but has only shown evidence of compensatory mechanisms for lameness from only one forelimb or hindlimb. The objective of this study was to create a consistent, controlled and immediately reversible lameness using a hoof clamp technique. The resultant changes in ground reaction forces measured by a force plate could then be measured in the normal and clamped limbs at a walk and trot. It would allow comparison of changes in forces in both the limb(s) in which lameness was induced and in the sound limbs among a variety of individual and paired-limb lameness scenarios to detect possible patterns in lameness and compensation. Lameness was induced in 8 clinically sound horses by tightening a circumferential hoof clamp on an individual forelimb, or hindlimb, a paired forelimb/hindlimb on the same side of horse, and a paired diagonal forelimb/hindlimb, and bilateral (both) forelimbs and hindlimbs. Force plate analysis was performed prior to (baseline) and after lameness induction. Using the hoof clamp technique, we were able to induce a consistent, controlled, and immediately reversible grade 2 out of 5 lameness at a trot in individual and paired limbs. In general, at a trot peak vertical force (force directed toward the ground), vertical impulse (force directed toward the ground times time), and braking impulse (braking force times time) decreased, and propulsion impulse (push off force times time) increased in the lame limb(s). The majority of compensation seemed to come from the limb directly opposite the lame limb. We demonstrated changes at the trot, but walk data remains to be analyzed. In a few random data sets that were examined, walking data appeared to change after the feet were clamped as well. The objective of the study for the summer scholar would be to examine the force plate data at a walk as well as the subjective videos and compare/contrast the findings to the data at a trot. Techniques and Applications the Student will Learn: Even though data has been previously collected, the student will learn how to look at equine lameness at the walk and trot using subjective (AAEP grading scheme) and objective measures (force plate). They will learn how kinetic data is generated and how to examine that data at the walk and trot (including graphical inspection and numerical, statistical comparisons). They will perform various data acquisition techniques from the raw data and then will learn how to assimilate and analyze the data and report it. Eligible Funding Sources: VPM
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Project Title: Novel Strategies and Tools for Monitoring Wildlife Health - Characterization of Wildlife Rehabilitation Data
Mentor: Dr. Michelle Willette
Contact: wille203@umn.edu
Co-Investigators: Dr. Andre Nault, Dr. Dominic Travis Project Description: Wildlife health is recognized as a key component of Ecosystem Health. Disease in wildlife has crucial implications for human health, food animal agriculture, and ecosystem services. Currently there is no comprehensive system for monitoring wildlife health in North America. However, there are more than 5,000 permits issued for wildlife rehabilitation, and it is estimated that more than 500,000 terrestrial and marine animals are seen at wildlife care centers on an annual basis. These wildlife care centers represent an untapped source of health data on a diverse array of wild animals, providing a unique ‘window’ into wildlife health. The Clinical Wildlife Health Initiative (CWHI) is an interdisciplinary collaborative working on the infrastructure and analytical tools required to utilize data from wildlife in rehabilitation settings. Clinical wildlife health data can provide novel and critical information that can be integrated into a national animal health monitoring infrastructure. Characterization of existing wildlife rehabilitation data is required to determine the accuracy and integrity of this information, as well as identify policy and knowledge gaps needed to utilize future data from wildlife care centers for surveillance and risk assessment. The specific objectives of this study are: 1. A Descriptive Review of Journal Articles in the Field of Clinical Wildlife Medicine: A systematic review of the scientific literature will be conducted and prepared for publication. The specific focus of the review will be to identify types of information supported by data from clinical wildlife medicine and gaps in the knowledge of how to apply information from clinical wildlife care centers to population level issues of human, animal, and environmental health. 2. An Analytical Review of Annual Federal Reports Submitted by Licensed Wildlife Rehabilitators Under the Migratory Bird Treaty Act: There are approximately 1500 wildlife rehabilitators permitted under the federal Migratory Bird Treaty Act who must file an annual report listing migratory birds by common name, date acquired, nature of injury, injuries resulting from potentially criminal activity, disposition, and date of disposition. Many of these rehabilitators are highly trained veterinary and/or conservation professionals. The Raptor Center has received copies of all annual reports from Region 3 (upper Midwest) for 2011. A systematic review of the reports will be conducted and prepared for publication. The specific focus of the review will be the scope and quality of data available in these reports. Techniques and Applications the Student will Learn: Scientific literature search; critical review and analysis of scientific literature; organizing and analytical assessment of data; wildlife epidemiology; risk assessment; public policy; scientific writing communication and dissemination of scientific information. An exceptional student can expect to be an author on two publications. Additionally, a summer scholar working on this project may spend 15-20 hours per week working in The Raptor Center Clinic obtaining experience in clinical procedures on raptors. Eligible Funding Sources: The Raptor Center Back to top Project Title: The role of baitfish in the dissemination of emerging viruses
Mentor: Dr. Nicholas Phelps
Contact: phelp083@umn.edu
Co-Investigator: Dr. Meggan Craft Project Description: The potential to inadvertently move aquatic animal pathogens to new regions through the movement of baitfish is a significant concern for fish health managers. Regulatory oversight has focused on high profile diseases, such as viral hemorrhagic septicemia, requiring annual inspections prior to movement for all susceptible species. However, many of the commonly sold baitfish species in Minnesota have not been found susceptible to VHSV, and as a result, continue to move undocumented. Numerous previously undescribed viruses have, however, been isolated from these fish since 2011. It is imperative that the movement of baitfish be modeled to demonstrate the potential distribution of emerging viruses. This information will directly influence natural resource management decisions and help inform a thorough risk analysis of the baitfish industry. Techniques and Applications the Student will Learn: Network modeling, industry and natural resource agency interaction Back to top
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