Summer Scholars Project Proposals

Summer 2018 Project Proposals

Comparative Biomedicine

Project Title: Applied behavior analysis: Helping animals cope and cooperate with medical care
Mentor: Melanie Graham
Contact: graha066@umn.edu

Project Description:

My lab is is a research unit of the Department of Surgery located on the veterinary hospital campus, primarily focused on translational research using complex nonhuman primate (NHP) disease models and screening rodent models. Consequently, 95% of animals in the center are enrolled in studies that measure behavioral, metabolic, immunologic factors or a combination of these. It is well understood that stress has a major measureable impact on each of these factors, which: introduces confounding, affects animal well-being, and increases number of animals used. The center is internationally recognized for developing successful behavioral management programs for animals used in research, with applications in husbandry and clinical care, and objectively demonstrated the value both in animal well-being and research outcomes.  The program fosters cooperation with caregivers, provides opportunities to express species typical behaviors, and promotes sociality.  Students will participate in behavioral management techniques aimed towards improving well-being of animals who undergo complex clinical care regimens.

Eligible Funding Sources: Boehringer Ingelheim, NIH T35, Department of Veterinary Population Medicine


Project Title: Targeting membrane stabilization in dystrophic muscle
Mentor: Joseph Metzger
Contact: metzgerj@umn.edu

Project Description:

Duchenne muscular dystrophy (DMD) is a devastating and universally fatal X-linked recessive disease of progressive striated muscle deterioration. DMD boys have severe skeletal muscle wasting early in life and they are typically wheelchair bound by age 10-12. DMD is fatal by the late teens to mid-twenties due to cardiac muscle and respiratory muscle failure. These grim realities underscore the tremendous urgency to discover and implement new approaches and therapies that could impact disease progression and outcomes in DMD. Presently, no effective clinical treatment that can halt or reverse DMD. Outstanding impact of this project is obtained by focusing on synthetic membrane stabilizers, in synergy with cell intrinsic membrane repair mechanisms, to directly target the primary physiological defect in DMD: severe muscle membrane instability. Membrane stabilizers confer marked protection to dystrophic cardiac and respiratory muscles, however, until now, protection of dystrophic skeletal muscles had not been shown, tempering enthusiasm in the DMD field for this class of therapy. The precise mechanism by which membrane stabilizers protect dystrophic muscles remains an open question and is tested here. Ultimately, an effective treatment must be applicable to all DMD patients, regardless of genetic mutation, and must treat all striated muscles: skeletal, respiratory and cardiac. The potential for significant impact of synthetic membrane stabilization, in conjunction with endogenous membrane repair, is thus multifold: 1) treatment efficacy is independent of the specific DMD disease gene defect: all DMD patients, regardless of genetic lesion, could be treated, 2) membrane stabilizers are chemically inert, non-immunogenic and non-metabolized molecule, thus negating the requirement for immunosuppression and 3) membrane stabilizer treatment shows evidence of protective effects for all striated muscle, skeletal, respiratory and cardiac muscles in DMD animal model studies in vivo. Mechanistic studies proposed here are enabled by our unique collaborative team of experts to decipher the fundamental basis of cell extrinsic/cell intrinsic membrane protection/repair. These studies are intimately connected with Project 1, Shared Resources and Training Cores to accelerate discovery. Guiding hypothesis: Synthetic membrane stabilizers will confer protection and synergize with cell intrinsic muscle membrane repair mechanisms to significantly enhance dystrophic skeletal, respiratory and cardiac muscle structure, function and viability in vivo. Aims include to investigate synthetic membrane stabilizers together with endogenous membrane repair mechanisms in preserving dystrophic limb skeletal muscles and respiratory muscle function and viability in vivo and to test synthetic membrane stabilizers and cell intrinsic membrane repair mechanisms in preserving dystrophic cardiac muscle function, viability and heart performance in vivo.

Eligible Funding Sources: Boehringer Ingelheim, NIH T35


Project Title: Animal models of osteochondrosis dissecans
Mentor: Ferenc Toth
Contact: ftoth@umn.edu

Project Description:

The research projects to which the summer scholar is welcome to join investigate osteochondorsis (OC) and osteochondrosis dissecans (OCD) in miniature pigs and horses. OC/OCD is a developmental orthopedic disease characterized by failure of the enchondral ossification in developing joints that often leads to secondary osteoarthritis.

The proposed equine study entails MRI of young foals with subclinical OC/OCD lesions in a private practice in Kentucky by our collaborators and subsequent evaluation of the MRI results here at the UMN. The aim of the study is to Identify and monitor progression vs. healing of subclinical OC lesions in young foals using bimonthly MRI exams and to describe specific markers of healing vs. progression. We hypothesize that T2 maps obtained in vivo are able to identify lesions of subclinical OC in foals and alterations in T2 relaxation time of OC lesions along with the size and location of the subclinical lesion can be used as predictors of lesion healing vs. progression.

The aim of the the study involving miniature pigs is to develop and validate an animal model that recapitulates all stages of juvenile OCD as it presents in young human subjects. We will test the hypothesis that a minipig model can be developed that closely reproduces human JOCD lesion progression, from its epiphyseal cartilage origin to late-stage clinical disease. This will be accomplished by surgically interrupting the vascular supply to the epiphyseal growth cartilage of the medial femoral condyle in juvenile miniature pigs. Operated animals will undergo serial MRI examinations to monitor the development and progression of osteochondrosis (OC) lesions. A subgroup of animals will be exposed to daily biomechanical trauma to facilitate development of clinically apparent disease (osteochondrosis dissecans, OCD). At the end of the study, animals will be euthanized, and operated and control femora will be harvested for histological evaluation.

Eligible Funding Sources: Boehringer Ingelheim, NIH T35


Project Title: Does normal microbial experience enhance MAIT cell numbers and function in mice?
Mentor: Kris Hogquist
Contact: hogqu001@umn.edu

Project Description:

Mucosal associate invariant T cells are a subset of T cells in the immune system that have innate or effector-like qualities. They are highly abundant in human blood, liver, skin, and GI tract. However, they are rare in mice, and have different effector properties, which limits the utility of mice as a model system. We hypothesize that laboratory mice have few MAIT cells because they lack normal microbial experience (being housed under specific pathogen free conditions). To test this, the scholar will analyze MAIT cells in "dirty" mice (laboratory mice co-housed with petstore mice). MAIT cells from various tissues will be identified using MR1 tetramers and analyzed by flow cytometry and for function in stimulation assay.

Eligible Funding Sources: Boehringer Ingelheim, NIH T35


Project Title: Role of Brain-Sympathetic-Gut Microbiome Axis in Hypertension
Mentor: John Collister
Contact: colli066@umn.edu

Project Description:

According to the CDC, hypertension affects 70 million American adults (~30%) and nearly 700 million people worldwide. It is the most significant risk factor for myocardial infarction and stroke, the first and third most frequent cause of death in Europe and the United States. Despite decades of research, the underlying cause of hypertension in most cases remains unknown. Furthermore, most afflicted individuals remain either untreated or suffer from ineffective treatment. It is our contention that new strategies are required to control high blood pressure and in the present project, our lab will move in an entirely new direction to examine the role of the gut microbiome on blood pressure regulation in two animal models of hypertension.

Our lab has demonstrated a role of the hypothalamic organum vasculosum of the lamina terminalis (OVLT), a circumventricular organ that has direct connections to the paraventricular nucleus to increase sympathetic activity, in the hypertensive response to both Angiotensin II (AngII) and DOCA (water soluble form of aldosterone) treated rats. We know 1) both of these models of hypertension have a sympathetic nervous system component, 2) the OVLT has a major role in mediating the hypertension, and 3) the gut is the most highly innervated peripheral organ with numerous sympathetic fibers. Taken together, we propose an OVLT-sympathetic-gut microbiome axis in the pathogenesis of hypertension.

In this project, we will examine the intestinal microbiome in two inducible rodent models of hypertension in OVLT lesioned and control rats to address the following AIMs: 1) What are the effects on the microbiome in chronic AngII and DOCA hypertensive rats and 2) Does OVLT lesion restore the microbiome while attenuating AngII and DOCA hypertension in the rat? In other words, the following hypothesis will be tested: OVLT lesion prevents the changes in gut microbiota and increased blood pressure in AngII and DOCA hypertension. In order to test this hypothesis, OVLT lesioned (or sham) instrumented rats will be treated with either AngII or DOCA. Continuous measurements of mean arterial pressure (MAP) and heart rate (HR) will be made via radio-telemetry. Rather than using fecal samples, direct microbiota analyses from jejunum, ileum, cecum and colon will be performed.

Eligible Funding Sources: Boehringer Ingelheim, NIH T35


Project Title: Genetic Investigation of a Juvenile Kidney Disease in Dogs
Mentor: Eva Furrow
Contact: furro004@umn.edu

Project Description:

Glomerulocystic kidney disease (GCKD) is an inherited disease characterized by cystic dilatation of the Bowman's capsules and atrophy of the glomerular tufts. Mutations in several different genes can result in GCKD in humans, and not all susceptibility genes have been discovered. In the veterinary literature, GCKD has been described in several breeds, but the genetic basis has not been reported. Our laboratory received DNA samples from three related Miniature Schnauzer puppies affected by GCKD. Pedigree analysis was consistent with an autosomal recessive inheritance. Whole genome sequencing was performed on DNA from one of the affected puppies, and the dog did not have a mutation in any of the genes known to cause GCKD in people. Thus, a novel susceptibility gene is suspected. The aim of this project is to perform a genome-wide association study in concert with further analysis of the whole genome sequencing data to uncover the responsible mutation. The results will be used to develop a mutation test to help Miniature Schnauzer breeders prevent future matings that could result in affected puppies. The data will also have translational value through the discovery of a new candidate gene for GCKD in people.

Eligible Funding Sources: Boehringer Ingelheim, NIH T35


Project Title: Mechanisms of anti-tumor immunity induced by oncolytic virotherapy for canine osteosarcoma
Mentor: Jaime Modiano, Co-PI: Kelly Makielski
Contact: modiano@umn.edu

Project Description:

Cancer therapy with oncolytic viruses is enjoying a renaissance. While maximizing the specific elimination of tumor cells, the contemporary emphasis is to understand how this therapy activates the immune system. We have initiated a clinical trial using vesicular stomatitis virus in dogs with osteosarcoma. The project will help to determine how this platform initiates anti-tumor immunity immunity against each dog's tumor, measuring molecular and cellular aspects of the response and its effects on survival.

Eligible Funding Sources: Boehringer Ingelheim, NIH T35, Skadron Family Fund


Project Title: Role of innate immune system in the progression of dystrophic cardiomyopathy
Mentor: DeWayne Townsend
Contact: town0045@umn.edu

Project Description:

The muscular dystrophies are a group of genetic diseases characterized by muscle wasting and degeneration. While the skeletal muscle is the most outwardly evident tissue affected, many of these patients also have significant cardiac disease as well. Duchenne muscular dystrophy (DMD) is the most common of the muscular dystrophies. This X-linked disorder is characterized by the loss of striated muscle cell membrane integrity in both skeletal muscle and cardiac tissues. While skeletal muscles have the ability to regenerate, cardiac myocytes do not. This low regenerative capacity of the heart places great importance on the factors defining cellular survival, especially after injury. The loss of membrane integrity results in the activation of an immune response characterized by extensive cellular infiltration. The nature of this infiltration and the effect these invading cells have on cell survival and tissue repair are poorly understood. The gap in our knowledge is important as modulation of the immune response following cardiac injury may represent a fruitful therapeutic strategy.

The central aim of this project will be to define the nature of the cellular infiltration into injured myocardium using a combination of immuno-fluorescent and flow cytometry.

Eligible Funding Sources: Boehringer Ingelheim, NIH T35


Project Title: Investigation of subclinical lesions of Juvenile Osteochondritis Dissecans (JOCD) in cadaveric human specimens
Mentor: Cathy Carlson, Co-PI: Ferenc Toth
Contact: carls099@umn.edu

Project Description:

This NIH-supported project focuses on the study of cadaveric tibiofemoral (knee) joints from children and uses CT, MRI, and anatomic/histological techniques to characterize the cartilage canal blood supply to the distal femur and determine if there are histological changes in sites of reduced blood supply that are similar to preclinical lesions of osteochondrosis dissecans in pigs. Previous pilot studies have confirmed that the blood supply to the distal femur in children and growing pigs is closely similar, and the documented predilection sites of juvenile osteochondritis dissecans (JOCD) and osteochondrosis dissecans (OCD) in both species are identical. The proposed work is designed to further investigate the association of areas of reduced cartilage canal blood supply with early/subclinical lesions of JOCD in children in order to better understand the pathogenesis of the disease and develop improved methods of early diagnosis and treatment.

Eligible Funding Sources: Boehringer Ingelheim, NIH T35


Project Title: Regulation of bone cells by estrogen induced myokines
Mentor: Kim Mansky, Co-PI: Dawn Lowe
Contact: kmansky@umn.edu

Project Description:

Osteoporosis and sarcopenia are maladies of aging and are worse in women than men. In recent years science has come to realize that bone and muscle are coupled not only mechanically but also at higher levels with biochemical and molecular signaling occurring between muscle and bone cells. However, how estrogen deficiency in females impacts the crosstalk between muscle and bone is not understood. We hypothesize that altered crosstalk between muscle and bone exacerbates the bone loss in females. This project is a collaboration between Dr. Kim Mansky (expertise in bone cells) and Dr. Dawn Lowe (muscle and estrogen expertise) to define muscle signals regulated by estrogen that regulate bone cells and the health of the skeleton. We will identify muscle signals by cytokine array and as well as expose both osteoclasts and osteoblasts to conditioned media to measure affects on differentiation and activity.

Eligible Funding Sources: Boehringer Ingelheim, NIH T35


Project Title: Investigating genetic bases for recurrent exertional rhabdomyolysis (tying-up) in horses
Mentor: Jim Mickelson, Co-PI: Molly McCue
Contact: micke001@umn.edu

Project Description:

Horses from many different breeds can suffer from the skeletal muscle disorder termed recurrent exertional rhabdomyolysis (RER), also commonly referred to as tying-up, in which an individual exhibits sporadic but recurring bouts of painful cramping and muscle cell damage following mild to moderate exercise. Although there can be many possible inciting causes, a large fraction of RER cases in Thoroughbred (TB) and Standardbred (STB) racehorses have genetic bases. A related condition exists in Quarter Horses (QH) where a condition now termed PSSM2 (for polysaccharide storage myopathy type 2) where several different genetic bases for tying up have been proposed. Current data indicates that up to 10% of the horses in each of these breeds have genetic susceptibility to RER/PSSM2.

A major goal of our equine genetic research group is to determine the genes and alleles that contribute to heritable musculoskeletal, neuromuscular and metabolic traits, such as RER and PSSM2. 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 with tens of thousands of DNA markers known as SNPs. From there, whole genome sequence data of cases and controls identifies the DNA sequence variants within the genes that reside in these regions. This state of the art research approach is used by human, veterinary, and other animal geneticists to define the genetic and molecular bases of many different types of heritable traits.

The major objectives of the project will be to use existing genome scan and sequence variant data to select and genotype plausible disease-causing variants in large case and control populations, and to analyze the data to determine the most-likely RER functional variants. Depending on progress made over the next 4 – 6 months, the student could be working on RER in TB, STB or PSSM2 in QH. Identification of the gene mutations underlying susceptibility to these disease will allow more precise definition of the disease pathophysiology, more accurate identification of susceptible horses, determination of the true prevalence of RER susceptibility, and help decrease its unknowing transmission to future generations.

Eligible Funding Sources: Boehringer Ingelheim, NIH T35


Project Title: Monitoring Immune Responses to Vaccine-based Immunotherapy in Dogs with Spontaneous Primary Brain Tumors
Mentor: Liz Pluhar, Co-PI: Mike Olin
Contact: pluha006@umn.edu

Project Description:

We have been treating pet dogs with spontaneously occurring primary brain tumors using autologous tumor lysate vaccine therapy for the past 7 years. We have demonstrated a peripheral immune response in dogs in a pilot study treated for meningiomas using Western blotting and cytotoxicity testing. This project will involve performing similar immune testing on stored serial blood and cell samples from dogs treated for primary meningioma or glioma of the brain.

Eligible Funding Sources: Boehringer Ingelheim, NIH T35, Skadron Family Fund


Project Title: Combined Immunotherapeutic Strategies for Glioma: Using Pet Dogs as a Large Spontaneous Model
Mentor: Liz Pluhar
Contact: pluha006@umn.edu

Project Description:

The objective of this proposal is to use pet dogs with spontaneous high-grade glioma to demonstrate the safety and efficacy of combination immunotherapy. In the current proposal, we will use our autologous tumor lysate and imiquimod vaccines in combination with our novel CD200 checkpoint blockade inhibitor or adenoviral-mediated gene therapy with Flt3L and TK in combination with our novel CD200 checkpoint blockade inhibitor to enhance immunotherapy for glioma. This project will include introduction to performing a clinical trial in pet dogs including recruitment, treatment (craniotomies and vaccinations), follow-up, and record keeping.

Eligible Funding Sources: Boehringer Ingelheim, NIH T35, Skadron Family Fund


Project Title: Cancer immunotherapies: Improving cancer cell killing by enhanced leukocytes
Mentor: Bruce Walcheck
Contact: walch003@umn.edu

Project Description:

Human and veterinary oncology face similar challenges, such as comparable incidence rates for certain types of cancer. Comparative basic and pre-clinical research is providing promising new breakthroughs for future cancer immunotherapies. An evolving immunotherapy for many types of cancer is the use of therapeutic antibodies that recognize tumor antigens. A key mechanism by which these antibodies kill cancer cells is leukocyte-mediated cytotoxicity. Our lab is using leukocytes derived from induced pluripotent stem cells and the peripheral blood to engineer them with improved receptors. These receptors are better at recognizing therapeutic antibodies and stimulating the leukocytes to kill cancer cells.

Eligible Funding Sources: Boehringer Ingelheim, NIH T35, Skadron Family Fund


Project Title:  Biology, assessment and therapy of acute and chronic pain in clinical and pre-clinical models
Mentor: Alonso Guedes
Contact: guede003@umn.edu

Project Description:

Our lab has several interesting projects evaluating the biology, assessment and therapy of acute and chronic pain in feline, equine and rodent models (either naturally occurring or induced models). Current projects include: 1) clinical evaluation of acute and chronic pain assessment tools in dogs and cats; 2) role of soluble epoxide hydrolase, a fatty acid metabolizing enzyme, in musculoskeletal pain in cats, horses and rodents; 3) Role of CD38 in nociception in rodent models.

Eligible Funding Sources: Boehringer Ingelheim, NIH T35


Project Title: Reprogramming the tumor and immune cell microenvironment using adrenergic receptor antagonists
Mentor: Erin Dickerson, Co-PI: Steve Jameson
Contact: edickers@umn.edu

Project Description:

Developing treatment approaches that simultaneously kill tumor cells while inducing potent immune responses has important implications for targeting both primary tumors and limiting recurrent disease. We recently showed that adrenergic receptor (AR) antagonists promote tumor cell death and enhance immune cell activation. These discoveries may explain the remarkable response rate of some cancer patients to treatment protocols using these drugs. Our laboratories are interested in identifying the mechanisms activated/inhibited by AR antagonists in order to generate more potent anti-tumor responses, and our studies have focused on metabolic pathways essential for tumor growth as well as T cell activation. The Summer Scholar will use in vitro metabolic assays to assess the activation of identified metabolic pathways in both tumor cells and T cells. In vivo studies will be used to determine whether AR antagonists alter these same metabolic programs to eradicate established tumors. These studies will provide proof-of principle regarding the efficacy of AR antagonists in combination with standard-of-care treatments for sarcomas. The Summer Scholar will work with researchers to develop an independent project and generate novel data.

Eligible Funding Sources: Boehringer Ingelheim, NIH T35, Skadron Family Fund


Project Title: Mutator Mice
Mentor: Reuben Harris
Contact: rsh@umn.edu

Project Description:

We have developed a novel strain of mice that expresses stably or inducibly a human DNA mutating enzyme. We are in the process of characterizing these animals with the goal of developing a new tumor model that better models human cancers. A summer scholar would have an opportunity to learn a number of experimental techniques and contribute to the development of this new cancer model.

Eligible Funding Sources: Boehringer Ingelheim, NIH T35


Project Title: Determination of key features of graft microenvironment that determine survival following transplantation
Mentor: Melanie Graham, Co-PI: Sabarinathan Ramachandran
Contact: graha066@umn.edu

Project Description:

Islet transplantation is considered a potential therapeutic option for selected patients with type 1 diabetes with history of hypoglycemia unawareness. Transplantation of isolated islets has been shown to ameliorate episodes of hypoglycemia unawareness and reduction in the secondary complications associated with type 1 diabetes. To avoid immunosuppression, tolerance has been a long-term goal in the field of transplantation. In a preclinical model, we have demonstrated that negative vaccination prior to islet transplantation along with a short course of immunosuppression induces tolerance and long term function of transplanted islets. In both the transplant setting as well as in cancer and other diseases there is evidence that the immediate microenvironment plays a crucial role in the activation of the immune responses. Likewise, it has also been shown by others that there are significant differences in the cytokine and chemokine profile in the immediate microenvironment when compared to the profile in peripheral circulation. Understanding the cytokine and chemokine signaling in the immediate microenvironment between tolerant and control groups would enable us to devise novel strategies to prevent the infiltration of the effector T cells to site of transplantation and promote the migration of regulatory immune subsets that can promote tolerance and long term survival of the transplanted islets. The aims of this study are: 1) Does the cytokine and chemokine profile in the portal circulation differ from that in peripheral circulation in normal animals? 2) Is the cytokine and chemokine profile in the portal circulation different in the control and tolerant animals? and 3) Can the cytokine and chemokine profile be used to predict the fate of the transplanted islets. In order to determine the differences in the cytokine and chemokine profile at the various compartments paired blood samples from the peripheral circulation and the portal vein will be evaluated. The cytokine and chemokine profile will be measured. The ability of the portal vein serum from the tolerant and control animals to induce migration of regulatory cells pr effector cells will be measured in a transwell migration assay. Further the ability of the serum to suppress the proliferation of donor specific T cells will be analyzed using a CFSE-MLR.

Eligible Funding Sources: Boehringer Ingelheim, NIH T35


Project Title: Brain bystander effects of radiation therapy in the treatment of solid tumors
Mentor: Davis Seelig, Co-PI: Jessica Lawrence
Contact: dseelig@umn.edu

Project Description:

Multimodal cancer treatment protocols combining chemo- and radiation therapy have improved the 5-year survival of cancer patients in the USA. However, an unintended consequence of these increasingly aggressive treatment regimens is the emergence of many unintended sequelae, including chemo- / cancer therapy-related cognitive impairment (CRCI), pulmonary fibrosis, and radiation-induced dermatitis. CRCI is a syndrome of neurocognitive decline characterized by decreased executive function, memory, attention and processing speed that affect up to 76% of patients undergoing cancer treatment and can persist for 5-10 years. Radiation therapy (RT) is a powerful and effective cancer treatment that is currently used in half of patients with solid malignancies. While it was long assumed that the biological effects of RT require direct interaction between beam and target cells / tissue, it is increasingly apparent that radiation can impact cells and tissues outside of the targeted area (including the brain). Based on human studies and previous mouse work in our laboratory, we hypothesize that these radiation induced bystander effects result in significant, persistent neuroinflammation that contributes to the development of CRCI.

The objective of this study is to further characterize the scope, consequences, and mechanisms underlying the impact of non-brain directed (NBRT) on the brain in both tumor-free and tumor-bearing mice. In these studies, we hypothesize that NBRT induces the production of inflammatory cytokines and mediators of oxidative stress within the radiation field that travel to the brain via either circulatory or neural routes to result in glial activation, neuronal dysfunction or death and neurocognitive impairment.

Specifically, this project will examine the neuropathologic and neurobehavioral consequences of NBRT’s brain bystander effects and the signaling mediators and conduits underlying these effects in cell culture systems and in tumor-free mice and tumor-bearing mice.

Eligible Funding Sources: Boehringer Ingelheim, NIH T35, Skadron Family Fund


 

Emerging, Zoonotic, and Infectious Diseases

Project Title: Development of novel live viral vaccines against significant veterinary diseases with infectious or non-infectious origins
Mentor: Hinh Ly, Co-PI: Yuying Liang
Contact: hly@umn.edu

Project Description:

We have recently developed a tri-segmented Pichinde virus (PICV) as a new vaccine vector platform to express up to two foreign genes (antigens) that induces strong humoral and cell-mediated immunity (Dhanwani et al., J Virol 2015). This vaccine vector can be administered via multiple delivery routes such as intramuscular, intranasal, subcutaneous as well as through oral gavage and induced strong immune response through each of the routes tested. In addition to the mouse model, we tested this vaccine vector in chickens and pig immunization experiments and demonstrated its efficacy in multiple species and through multiple routes of immunization. No lateral transfer of the viral vector was observed when the immunized animals were co-housed with naïve animals. The vaccinated animals did not develop any adverse effects attributable to immunization. These factors attest to the good safety profile of this new vaccine vector platform. We have recently obtained new funding from multiple sources, including the Minnesota Agricultural Experiment Station and the US-Israel bi-national agricultural research and development fund, to use this new viral vector platform to develop vaccines in order to protect food-production animals (e.g., poultry and swine) against significant diseases with infectious origins. We are also interested in developing novel vaccines against communicable and noncommunicable diseases afflicting companion animals wherever applicable.

Eligible Funding Sources: Boehringer Ingelheim, NIH T35


Project Title: Social network analysis and disease transmission in free-ranging mountain lion (Puma concolor) populations
Mentor: Meggan Craft, Co-PI: Marie Gilbertson
Contact: craft@umn.edu

Project Description:

Disease transmission within animal populations is affected by contact frequency and social structure, often represented as contact networks. Few studies are able to assess contact networks within wildlife, and even fewer do so longitudinally. The goal of this project is to assess contact networks of mountain lion populations in study areas from California, Colorado, and Florida across time spans of up to 10 years using existing telemetry data. These different study areas experience different levels of anthropogenic land-use change and different wildlife management strategies, resulting in variations in habitat fragmentation and population densities. By performing longitudinal analysis of changes in contact network structure and social network analysis, we hope to better understand how habitat fragmentation and changes in population size may affect network connectivity. These findings have implications for disease transmission across populations and will be important for understanding how habitat fragmentation and changes in population size affect disease risk in wildlife.

Eligible Funding Sources: Department of Veterinary Population Medicine


Project Title: Optimization of a highly sensitive real-time PCR and evaluation of pooled turkey gastrocnemius tendons for detection of turkey arthritis reovirus (TARV) by PCR.
Mentor: Sunil Kumar, Co-PIs: Rob Porter, Sagar Goyal
Contact: kumars@umn.edu

Project Description:

Justification: Turkey arthritis reovirus (TARV) has been characterized by our VDL research group. TARV is a novel virus, distinct from chicken reovirus, that causes severe lameness in 12-18-week-old commercial turkeys. The virus is significant in turkey production because it affects both animal wellbeing and has causes substantial market losses due to culling. Virus isolation in embryonated eggs and QT-35 cell line is the gold standard for detection of turkey arthritis reovirus. Real-time PCR (rRT-PCR), has a sensitivity of 65%, but can provide a positive result to the turkey producer in a much shorter time than virus isolation. We hypothesize that: (1) tendon needs special processing for getting good results (improved sensitivity); (2) Individual leg tendon testing or pooling less no of legs is better than pooling tendons from five or more legs. Currently, the detection of TARV by random collection of leg tendons from multiple birds has been the standard practice. There is no standard of how many tendons are required (minimum or maximum) for successful virus isolation and PCR in positive cases. We currently recommend to collect and pool samples from 5-6 turkey legs, but this can be rather cumbersome and expensive for producers to ship heavy legs to the MVDL in St. Paul.

Aim: To optimize tendon processing and to determine if the number of legs tested for TARV can be reduced without affecting the test outcome for virus isolation and PCR.
Procedures: During the next three months individual tendon samples will be collected from turkey lameness cases containing 3-6 legs. Tendon samples (1cm segment of gastrocnemius tendon and 1 cm segment of digital flexor tendon) from each leg will be collected, pooled, and stored at -70oC. Samples will be processed by different methods and rRT-PCR will be done on pooled samples from one leg and the cT value will be recorded. Samples from 3 and/or 5 legs will then be pooled and subjected to rRT-PCR. The cT value of 3/5 pooled legs will be compared with those of a single leg by analysis of variance. This will also identify the optimum testing method and whether testing of individual legs yields the same result as testing pooled samples from multiple legs.
Expected Outcome:
Results will improve sensitivity of current assay and reduce the number of samples required for collection by the veterinarian or turkey producer.

Eligible Funding Sources: Department of Veterinary Population Medicine


Project Title: Effect of small-colony variants phenotype on growth kinetics, intracellular persistence and infection of Salmonella enterica Enteritidis
Mentor: Sinisa Vidovic
Contact: svidovic@umn.edu

Project Description:

Under certain circumstances, the acquisition of antimicrobial resistance has been associated with the development of small-colony variants (SCVs), a bacterial phenotype characterized by the formation of profoundly small colonies compared to their parental counterparts. It has been shown that development of SCVc very often is associated with the field antimicrobial treatment due to the ability of SCVs to reduce metabolism.
Recently, we obtained two groups of spontaneous SCVs of Salmonella Enteritidis that have acquired resistance (4 ug/mL) and high resistance (40 ug/mL) to ciprofloxacin, respectively compared to their parental strains that have exhibited a susceptible (0.05 ug/mL) phenotype to the same drug.
We plan to initiate a research project to determine the effect of SCVs phenotype on the fitness of Salmonella Enteritidis. In particular, we are going to determine growth kinetics, intracellular persistence and infection potential of SCVs compared to that of their parental strains.

Eligible Funding Sources: Boehringer Ingelheim, Department of Veterinary Population Medicine


Project Title: Developing GMR based tests for Mycoplasma hyopneumonia
Mentor: Maxim Cheeran, Co-PI: Maria Pieters
Contact: cheeran@umn.edu

Project Description:

The Pork industry has an annual gross production output of ~$20 billion (US Census of Agriculture, 2012) and respiratory diseases affecting up to ~50% of the herd at any given stage of production have significant economic outcomes. Rapid identification of pathogen(s) involved in respiratory diseases at the farm could facilitate efficient implementation of control and prevention strategies that curtail disease outbreaks in swine production systems. a Giant Magnetoresistance (GMR)-based biosensor chip for detecting three PRDC pathogens using functionalized magnetic nanoparticles (MNP), utilizing the property of the biosensor to detect minute changes in magnetic fields.

One of these pathogens is Mycoplasma hyopneumonia, a pathogen for which standard laboratory tests have limited value or are inaccessible to the producer. We propose to develop both rabbit polyclonal and mouse monoclonal antibodies (MAb) to M. hyopneumoniae, reference strain 232. ELISA and western blotting techniques will be used to evaluate the specificity of antibodies by comparing binding affinities to the reference strain (232) and to related Mycoplasma sp. such as M. flocculare, M. hyosynoviae, and M. hyorhinis. Cross-reactivity to other respiratory pathogens including Streptococci, Pasteurella, Actinobacillus, PRRSV and SIV will be evaluated prior to selecting antibodies for GMR sensors. MAbs and rabbit polyclonal antibodies with the desired specificity will be purified and used to develop capture and detection reagents, respectively, for a GMR biosensor-based assay.

Eligible Funding Sources: Department of Veterinary Population Medicine


 

 

Population Systems

Project Title: Evaluating the impact of a lameness intervention strategy
Mentor: Gerard Cramer
Contact: gcramer@umn.edu

Project Description:

Lameness is a painful and prevalent disease in the dairy industry. Despite our knowledge and understanding, lameness persists as an unbounded industry problem with far-reaching economic and cattle welfare consequences. In order to meaningfully reduce the prevalence of lameness we must motivate and support producers to successfully make on-farm changes. Our objective for this project is to develop, implement and evaluate the participatory and team-based Healthy Steps Lameness Control Program. This program aims to create an environment that builds producer collaboration and allows for peer advising and support by training veterinarians/hoof trimmers/nutritionists to facilitate meetings with small groups of producers. Meetings will include in-class activities, on-farm tours, and case-studies. The program will be assessed using a mixed methods approach of quantitative risk assessment scores and qualitative participant interviews in order to yield a fuller understanding of lameness control decisions and participant experience.

Eligible Funding Sources: Dairy Group, Department of Veterinary Population Medicine


Project Title: Molecular techniques for studying the epidemiology of meningeal worm
Mentor: Tiffany Wolf
Contact: wolfx305@umn.edu

Project Description:

Parelaphostrongylus tenuis is one of the leading causes of natural moose mortality in Minnesota. This natural parasite of white tailed deer is transmitted to moose via a gastropod intermediate host, but questions remain about where in their natural habitat moose become infected. My research group is developing molecular approaches to answer this question, and there is much baseline work needed to validate our molecular techniques. We are seeking a creative student to help design and conduct experiments to: 1. examine the extent of within-host genotype heterogeneity that exists from larvae shed by deer (since sexual reproduction takes place within the deer host); and 2. validate qPCR as a tool to quantify shedding rates in feces and larval load in gastropods. The student will work along with others in my lab and a number of colleagues in the CVM and the UMN Genomics Center, as well as work independently in the laboratory to recover larvae from deer pellets and set up the proposed experiments. The motivated student will also lead the development and submission of a manuscript resulting from this work for publication in a relevant journal.

Eligible Funding Sources: Department of Veterinary Population Medicine


Project Title: To study the respiratory virome of backyard poultry and to educate owners about respiratory pathogens
Mentor: Rob Porter, Co-PIs: Sunil Kumar, Sagar Goyal
Contact: porte349@umn.edu

Project Description:

Small flock owners (SFOs), including backyard flock owners, need to learn about common pathogens that may affect their birds and how to control them. After the HPAI (highly pathogenic avian influenza) outbreaks in 2015, we at the Minnesota Veterinary Diagnostic Laboratory (MVDL) have seen an increase in number of cases submitted and number of phone calls received from backyard poultry owners; they are nervous and believe that any sign of respiratory problem in their flocks is due to HPAI although in a large majority of such cases we detected other respiratory pathogens e.g., infectious laryngotracheitis virus (ILTV), infectious bronchitis virus (IBV), Newcastle disease virus (NDV), and Mycoplasma gallisepticum (MG); either alone or as mixed infection. A comparison of respiratory viromes of clinically affected and normal chickens will help delineate the viral load in their upper respiratory tracts and this information will help formulate strategies to control these infections. This information can then be communicated to the flock owners along with advice on vaccine usage, if applicable. With this background, we hypothesize that respiratory virome of backyard chickens contains signatures of common and novel pathogens and that the lack of knowledge on these pathogens leads to a panic situation among SFOs. To fulfil the objective of Poultry Respiratory Disease-Coordinated Agricultural Project (PRD-CAP), we plan to direct extension efforts towards two audiences: SFOs and veterinarians (both mixed practice and companion animal veterinarians) who often are not familiar with the prevalence and control of poultry diseases. We are working now to develop educational materials, which will help this audience in recognizing respiratory infections, how to use the veterinary diagnostic laboratory effectively, and how to prevent the introduction of respiratory diseases in poultry flocks (including biosecurity and vaccination programs). Practicing veterinarians will receive continuing education on backyard poultry and their health. To develop these educational materials, we will use a variety of tools (eXtension webinars, workshops, short videos, disease recognition app, brochures, and fact sheets). Our team is also working on preparing a user manual containing lectures on respiratory diseases with images of clinical signs, control and prevention of such diseases. This manual will be distributed to all participants and will also be made available at University of Minnesota Extension website as well as at the MVDL website. The study will enhance outreach efforts to control and prevent respiratory diseases in backyard poultry, which may also benefit commercial flocks and help reduce zoonoses.

Eligible Funding Sources: Department of Veterinary Population Medicine


Project Title: Critical review measures listed in the international (Chile, Norway, Scotland, Faroe Islands, Canada, US) legislation limiting and framing the location and activities of salmon reproduction centers in the sea.
Mentor: Andres Perez, Co-PIs: Alex Primus, Amy Kinsley
Contact: aperez@umn.edu

Project Description:

A systematic review, complemented by web-searches, and active contact of key partners in those countries will be conducted to collect, organize, and critically review the related legislation in countries in which salmon production is relevant. As part of the critical review, changes that some are proposed to the Chilean legislation will be put into context. Scientific literature that assess the efficacy of legislative change will be collected and summarized. Experts will be contacted to gather information and evaluate consensus on the appropriateness of implementation of key measures and explore reasons for potential differences.

Eligible Funding Sources: Department of Veterinary Population Medicine


Project Title: Development of a nanoparticle-based oral vaccine strategy for fish
Mentor: Alex Primus, Co-PIs: Sunil Mor, Chun Wang
Contact: primu012@umn.edu

Project Description:

Aquaculture is one of the fastest growing food production systems worldwide and the importance of its role in providing food security to a rapidly expanding global population is expected to increase significantly in the next several decades. The two most significant issues that have been identified as potentially limiting factors for the success of aquaculture in the near-future are: 1) The ability to effectively control infectious disease; and, 2) Ensuring that production methods are environmentally sustainable. Vaccination of fish in aquaculture could play a key role in managing both of these challenges.

While the use of vaccines in aquaculture has successfully helped prevent disease in some fish species over the last couple decades, there are still several aspects of aquaculture vaccines that could be greatly improved. One area in which there is a particular need for improvement is related to the administration method of efficacious vaccines. The most efficacious vaccines currently available for fish are those that are administered by injection of each individual fish (most of which are injected intraperitoneally). Given the high costs associated with injecting each individual fish, administering vaccines to fish by injection is only economically justifiable for a few high-value fish species such as Atlantic salmon. Consequently, the aquaculture industry has been pursuing the development of vaccines that can be administered either orally or by immersion. Most oral and immersion vaccine preparations, however, lack the efficacy and duration of immunity provided by injectable vaccines. For the oral vaccines, this lack of efficacy is thought to be largely due to digestion of the antigen in the stomach prior to entry into the intestine where it can be absorbed to stimulate an appropriate immune response.

In the current project, we propose investigating the development of a relatively novel oral vaccine strategy for fish using nanoparticles. This strategy, variations of which have been investigated for humans, other mammals, and some fish species, involves the microencapsulation of antigens of interest in synthetic nanoparticles. Encapsulating the antigens effectively protects them from degradation in the stomach, allowing them to reach the intestine where they may be absorbed to produce a protective immunity. Initial steps in this process will involve developing stable nanoparticles containing our antigen, incorporating these nanoparticles into feed, administering this feed to fish, and evaluating the localization of antigen in the intestine and/or checking for a humoral response.

Eligible Funding Sources: Department of Veterinary Population Medicine


Project Title: Biosecurity approaches for swine farms
Mentor: Montse Torremorell
Contact: torr0033@umn.edu

Project Description:

Protecting the health of animals is a priority for swine producers. There are two areas of research as part of this project. The first area has to do with developing methods to validate biosecurity practices for swine farms such as the use of UV boxes for decontaminating fomites that enter the farms. The second area includes the discipline of aerobiology and has to do with developing and validating analytical methods to detect PRRSV in filters and investigate PRRSV breaks in filtered farms.

Eligible Funding Sources: Department of Veterinary Population Medicine


Project Title: Investigating the effect of time in the calf hutch post weaning on subsequent health and performance in dairy calves
Mentor: Whitney Knauer, Co-PI: Sandra Godden
Contact: knaue020@umn.edu

Project Description:

The successful transition from milk to solid feed is important for the growth and health of young dairy calves. One way that producers encourage grain intake is to keep calves in hutches past the cessation of milk, so that individual intake can be monitored and that competition for resources with other animals is limited. However, recommendations for the duration of time that calves should be kept in hutches has not been studied. Additionally, little information exists to describe the hutch microenvironment as the calf grows and becomes a ruminant, which could be important for subsequent health and performance. The objective of this trial is to investigate the effect of time in the hutch post weaning and subsequent health and growth in dairy calves. A secondary objective will be to describe the hutch microenvironment and its associations with calf health and growth.

The student would conduct this research in a field laboratory setting on a large commercial dairy farm in MN or TX. In addition to conducting research activities, students would have a fantastic opportunity to get involved with other day to day activities on the dairy, including palpation, sick cow management, and obstetrics. The student would have the opportunity to present the results of their project at the annual meeting of the American Association of Bovine Practitioners in Fall, 2019.

Eligible Funding Sources: Department of Veterinary Population Medicine

 


Project Title: Occurrence of hyperketonemia in subsequent lactations
Mentor: Luciano Caixeta, Co-PI: Gerard Cramer
Contact: lcaixeta@umn.edu

Project Description:

The objective of this study will be to determine the likelihood of the occurrence of hyperketonemia in subsequent lactations. Hyperketonemia is defined by elevated blood concentrations of β-hydroxybutyrate (BHB > 1.2mmol/L) and have been associated with disease occurrence, decreased milk production, poor reproductive performance, and increased culling rates. Even though, diet and milk production greatly affect the levels of BHB in circulation during early lactation, a recent study showed that genetic differences are associated with different hyperketonemia incidence. Considering the genetic component of hyperketonemia, we can hypothesize that animals diagnosed with hyperketonemia in the previous lactation are more likely to have elevated BHB concentration in a subsequent lactation. However, the occurrence of hyperketonemia in consecutive lactations has not been investigated in dairy cows to date. Thus, in this study, BHB concentration during early lactation will be determine for two consecutive lactations. In the summer of 2017, BHB concentration during early lactation was measured by our research group in approximately 4,000. Based on calving interval and culling rate in commercial farms, we expect that over 2,500 animals will still be in the herd and starting a new lactation; thus, eligible to have BHB measured for the second time. Animals will be kept under normal conditions at their home herd and blood samples will be collected at week 1 (7 ± 3 days in milk – DIM) and week 2 (14 ± 3 DIM) on a weekly basis. The blood BHB concentration will be measured using a cow-side ketone meter (Centrivet GK, Acon Laboratories, San Diego, CA). Animals will be followed during the first 60 DIM to determine the incidence of diseases. To our knowledge, this will be the first study reporting the incidence of hyperketonemia in subsequent lactations. This proposed work will generate knowledge on the importance of the genetic component of hyperketonemia in dairy cows.

Eligible Funding Sources: Department of Veterinary Population Medicine