Summer Scholars Project Proposals

Summer 2019 Project Proposals

Comparative Biomedicine

Comparative Biomedicine

Project Title: In vitro and in vivo Evaluation of Induced Pluripotent Stem Cell Derived Chondrocytes
Mentor(s):  Toth and O’Brien
Contact:  ftoth@umn.edu

Project Description: Cartilage tears and osteochondral defects are devastating injuries that often lead to osteoarthritis. Currently available treatment options for cartilage lesions are expensive, and frequently fail to restore joint function. Novel therapies that can improve clinical outcomes over current methods are therefore needed. In recent years, cell-based therapies, such as autologous chondrocyte implantation (ACI), have gained interest for the treatment of cartilage defects. ACI has been associated with good results in 60-90% of patients but its wider adoption is hampered by the morbidity and cost of the additional surgical procedure required to harvest autologous chondrocytes for culturing. Thus, other sources of cells that are easily accessible and have the potential to restore articular cartilage are of great interest. One option entails utilization of induced pluripotent stem cells (iPSCs) as a source of chondrocytes. We have recently discovered a new method to induce differentiation of hyaline cartilage from iPSCs via the initial formation of brain organoids. We will evaluate the dynamics of cartilage formation in iPSC-derived organoids over time and demonstrate the presence of hyaline cartilage using RNA sequencing and immunohistochemistry (Aim 1). Secondly, we will test the ability of organoid derived chondrocytes to facilitate repair of induced cartilage defects in vivo in a goat model (Aim 2).
The primary impacts of the proposed study will be characterization and in vivo evaluation of a novel source of chondrocytes for cell-based cartilage repair strategies that will lay the foundation for a future one stage cartilage repair technique.

Techniques Used: Experimental surgical procedures in goats,
post operative management, MRI of the experimental animal,
post mortem processing and evaluation of joint specimens

Eligible Funding: Boehringer Ingelheim, NIH T35, Dept. Veterinary Clinical Sciences


Project Title: Aging and Osteoarthritis
Mentor(s): Carlson and Armstrong
Contact: carls099@umn.edu

Project Description: Osteoarthritis is a common, debilitating, and costly disease affecting both animals and human beings. The proposed work is aimed at better understanding the pathogenesis of this condition through the use of mouse models of naturally occurring and surgically-induced disease. The live animal work for these studies is done at another institution; work done at the U of MN includes producing, staining, and evaluating histological sections of murine stifle joints using an osteoarthritis grading scheme and taking histomorphometric measurements of cartilage and bone features. This project offers a great learning opportunity for students interested in orthopaedics, particularly osteoarthritis.

Techniques Used: Embedding tissues in paraffin; cutting histological sections using a microtome; staining sections and coverslipping the glass slides; grading sections using a specialized scheme designed to evaluate osteoarthritis in mice; using an OsteoMeasure histomorphometry program to measure cartilage and bone areas

Eligible Funding: Boehringer Ingelheim, NIH T35, Dept. Veterinary Clinical Sciences


Project Title: Neuroimmunology of Traumatic Brain Injury
Mentor(s): Cheeran, Grande, and Low
Contact: cheeran@umn.edu

Project Description: Known as the ‘silent epidemic’, traumatic brain injury (TBI) is the leading cause of mortality and disability in those under the age of 45 in the USA. Acute inflammation after TBI has long been recognized for its role in recovery but is thought to resolve over time. Increasing evidence of activated microglia have been found in the brain years after TBI, in human autopsy studies and more recently replicated in animal models. These chronically activated and ‘primed’ microglial/macrophage mount a hyperactive inflammatory response to subsequent stimuli such as TBI, which may explain the persistent and evolving neurodegeneration in humans and animals after repetitive TBI. In this project we will test the ability of non-hematopoietic umbilical cord blood stem cells (nh-UCBSC) to modulate the macrophage-induced inflammation following TBI. the short and long-term changes will be evaluated by measuring the extent, distribution, and temporal evolution of TBI-induced neuro-inflammation. Current treatment of TBI is aimed at preventing secondary complications. Unfortunately, there is no treatment available to reverse or stop the deleterious neuroinflammation that occurs after brain injury. The novel cellular therapy with nh-UCBSC has already been shown to do alter the inflammation in stroke where similar neuroinflammatory processes exist. If successful, our cellular therapy would be a paradigm shift in the management of TBI. Not only would this be expected to prevent tissue loss and brain damage shortly after TBI but also prevent long-term sequela such as chronic traumatic encephalopathy.

Techniques Used: Flowcytometry, immunohistochemistry, controlled coritcal impact model, bone marrow chimeras and evaluation of neuroinflammation by measuring cytokine responses, and immunophenotyping immune responses in the brain.

Eligible Funding: Boehringer Ingelheim, NIH T35, Dept. Veterinary Population Medicine


Project Title: Role of Gut Microbiome in Two Models of Hypertension
Mentor(s): Collister
Contact: colli066@umn.edu

Project Description: We have 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. 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?;  2) Does OVLT lesion restore the microbiome while attenuating AngII and DOCA hypertension in the rat?;  and 3) Does ileal transplantation from OVLT lesioned rats prevent or treat 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, and cross-transplanted with ileal content during the hypertensive treatment. 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.

Techniques Used: Training will begin with the surgical procedures involved with chronic instrumentation of the rat. Students will learn placement of a chronic indwelling femoral IV and ileal catheters, and placement of the blood pressure transducer catheter in the abdominal aorta, as well as anesthesia, general surgical aseptic technique and monitoring of the surgical patient during anesthesia. Students will learn daily metabolic measurements of rodents housed in metabolic cages, computer data compilation of continuous collections of heart rate and blood pressure data via radio telemetry. Lastly, students will learn the proper techniques of euthanizing experimental rats, and harvesting brain and gut tissues for histological and microbial genomic analyses, respectively.

Eligible Funding: Boehringer Ingelheim, NIH T35, Dept. Veterinary & Biomedical Sciences


Project Title: Genetic Investigation of Rare Urinary Stones in Exotic Species
Mentor(s): Furrow and Lulich
Contact: furro004@umn.edu

Project Description: In conjunction with the Minnesota Urolith Center, we have identified rare hereditary urinary stones in multiple exotic species. Examples include cystine stones in ferrets and Fennec foxes and xanthine stones in otters. In some cases, the stones resulted in fatal kidney disease as kits/pups. The aim of the proposed summer scholar research is to perform candidate gene sequencing to discover the mutations underlying risk for these hereditary stone disorders. The results are critical to the health of each affected species. Once mutations have been discovered, tests can be designed to screen breeding stock and thereby prevent the production of affected animals.

Techniques Used: PCR, electrophoresis, DNA sequencing, variant analysis, comparative genomics

Eligible Funding: Boehringer Ingelheim, NIH T35, Dept. Veterinary Clinical Sciences, Morris Animal Foundation


Project Title: An in vitro Porcine Blood Brain Barrier Model to Differentiate Virulent and Commensal Strains of Streptococcus suis
Mentor(s): Gebhart
Contact: gebha001@umn.edu

Project DescriptionStreptococcus suis is an increasingly significant cause of meningitis-related deaths in piglets. Highly virulent strains are also associated with arthritis, endocarditis, and septicemia. Serotyping and multi-locus sequence typing (MLST) have identified several high-virulence S. suis subtypes in Europe, as there are few subtypes responsible for most clinical cases there. Our project is to investigate approaches that can differentiate pathogenic from commensal strains in vitro to select S. suis isolates for use in challenge trials or as vaccine candidates. We propose evaluating a porcine blood brain barrier (BBB) model for evaluating interactions of S. suis strains of various pathotypes and virulence-associated gene profiles with brain endothelial cells and astrocytes.  S. suis must transverse the BBB to ultimately cause inflammation and meningitis. Brain microvascular endothelial cells and astrocytes are two major cell types that compose the blood brain barrier and this co-culture model provides a more complex environment for studying pathogen-cell interactions (e.g. adherence, invasion) than monocultures.
For this project, we will establish an in vitro porcine BBB model and perform cell invasion and adherence assays to identify characteristics of highly virulent reference strains as compared to commensal strains of S. suis. This model will be used to compare the adherence and invasion by various known or putative virulent and commensal strains of various serotypes and sequence types.

Techniques Used: Bacterial culture, cell culture, cell invasion and adherence assays, PCR, MLST

Eligible Funding: Boehringer Ingelheim, Dept. Veterinary & Biomedical Sciences


Project Title: Prevalence of Performing Lymphadenectomy with or without Sentinal Lymph Node Mapping for Prognostic and Therapeutic Treatment of Tumors in the Head and Neck : a Multi-institutional Study
Mentor(s): Goldschmidt
Contact: golds245@umn.edu

Project Description: Tumor staging including evaluation of locoregional draining lymph nodes and distant sites for presence of metastasis is a critical step in the oncologic work up. The ideal lymph node to sample for presence of metastasis is the sentinel lymph node. The sentinel lymph node is defined as the first lymph node within the lymph node basin that drains the primary tumor. In humans, where sentinel lymph node identification is standard practice for several histologic types, it has been shown that if the sentinel lymph node is negative for metastasis, then additional draining lymph nodes are also negative in over 90% of cases. Thus, pathology of the sentinel lymph node is vital information to acquire. However, sentinel lymph node mapping is in its infancy in veterinary medicine. Thus, many institutions are currently removing both the mandibular and retropharyngeal lymph node bilaterally in all cases of oral tumors, which can carry significant morbidity. This study would identify what institutions are removing all lymph nodes, versus lymph nodes identified as "sentinel" on imaging, what complications they are encountering, and how the information is changing their clinical practice.

Techniques Used: Literature review, survey design, collating results, and performing statistical evaluation if possible

Eligible Funding: Dept. Veterinary Clinical Sciences, Skadron Family Scholarship


Project Title: Role of iNKT Cells in Pathogen Protection of the Developing Fetus
Mentor(s): Hogquist
Contact: hogqu001@umn.edu

Project Description: Lipid specific T cells are conserved throughout mammalian evolution. Despite being widely studied, it is unclear what essential functions these T cells (called invariant natural killer T cells or iNKT cells) serve. To understand this better, we exposed mice lacking iNKT cells to the microbes that are naturally occurring in mice through co-housing with pet store mice. While adult animals were healthy, developing pups lacking CD1d did not survive. This project will examine iNKT cells in the placenta and explore how iNKT cells protect the developing fetus.

Techniques Used: Flow cytometry, viral infection of animals, in vitro assays of immune functions (cytotoxicity and cytokine production), possibly RNAseq and immunofluorescence as well.

Eligible Funding: Boehringer Ingelheim, NIH T-35, Skadron Family Scholarship


Project Title: Genetic Determinants of Pediatric Solid Tumor Metastasis
Mentor(s): Largaespada
Contact: larga002@umn.edu

Project Description: My focus is on genetics, gene modification and tumor biology. Our group performs in vivo forward genetic screens for tumor development and metastasis in mice using the Sleeping Beauty transposon. Using this method, we’ve identified new genes and genetic pathways that cause colorectal cancer, osteosarcoma, hepatocellular carcinoma, leukemia, malignant peripheral nerve sheath tumors, and brain tumors. The findings are being used to test new therapies for osteosarcoma, and brain and peripheral nerve sheath tumors. We’re also using small molecule and genetic (CRISPR/Cas9) screens to find selective drug sensitivity or synthetic lethal relationships with loss of certain tumor suppressors. A special focus of our work are the benign and malignant tumors that develop in the context of the cancer predisposition syndrome Neurofibromatosis Type 1.

Techniques Used: Tissue culture, CRISPR/Cas9, mouse husbandry

Eligible Funding: Boehringer Ingelheim, NIH T35, Skadron Family Scholarshi


Project Title: Innovations in Cancer Immunotherapy
Mentor(s): Modiano
Contact: modiano@umn.edu

Project Description: Recent advances are helping us to understand how we can use the immune system to treat cancer safely and effectively. We have developed a novel drug, called eBAT, which consists of recombinant epidermal growth factor and urokinase linked to Pseudomonas exotoxin. eBAT has shown efficacy in the treatment of spontaneous canine hemangiosarcoma. One of its potential mechanisms of action includes targeting of myeloid immunosuppressive cells, potentially leading to re-activation of anti-tumor immunity in cancer patients. A complementary approach to activate the immune system is to disrupt the local tumor environment to create productive inflammation. This project will explore mechanisms to activate the immune system using eBAT and/or tumor disruption using laboratory animal models.

Techniques Used: Cell culture, microscopy, flow cytometry, cell killing assays, lab animal (mouse) handling and husbandry, and methods to track antigen specific tumor responses in vivo. In addition to project-specific methods, every student will be trained in routine laboratory safety, ethics in research, teamwork, and written and oral presentation.

Eligible Funding: Boehringer Ingelheim, NIH T35, Skadron Family Scholarship, Morris Animal Foundation, Dept. Veterinary Clinical Sciences


Project Title: Mechanisms of Radiation Induced Side Effects During Cancer Therapy
Mentor(s): Seelig and 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. Radiation therapy (RT) is currently recommended in more than 50% of patients with solid malignancies. Unfortunately, adverse events are common with radiation therapy and may negatively affect quality-of-life. Our research group investigates key changes that occur when normal tissue is irradiated, to help develop new treatments that mitigate unwanted toxicities such as 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. Pulmonary fibrosis can develop years following radiation treatments for head and neck cancers and lung cancers and may affect lung function. Radiation induced dermatitis, characterized by erythema, moist desquamation and pain, is an acute radiation toxicity that occurs in over 50% of patients who undergo curative-intent radiation therapy. The biological effects of radiation can occur within the treated field or, as is the case in CRCI or lung fibrosis, can affect cells and tissues outside of the targeted area. We hypothesize that these adverse effects of RT result from significant, persistent inflammation and the increased production of pro-inflammatory cytokines, and the creation of reactive oxygen species that injure unirradiated cells. Therefore, the overarching objective of our research is to explore the mechanisms underlying the adverse effects profile during and after RT. In CRCI, we are principally interested in the role of microglia, including their activation profile in response to directed and bystander radiation. In pulmonary fibrosis, we are interested in evaluating serial changes that occur in irradiated lung over time after radiation exposure. In radiation-induced dermatitis, we are engaged in the optimization of a mouse model of this condition so as to define the clinical, microscopic (pathologic), and molecular responses to direct RT injury.

Techniques Used: Flow Cytometry, RT-PCR, Immunohistochemistry, Immunocytochemistry

Eligible Funding: Boehringer Ingelheim, NIH T35, Dept. Veterinary Clinical Sciences, Skadron Family Scholarship


Project Title: Design and Optimization of a Flow Cytometric Analysis Platform for Canine Bone Marrow
Mentor(s): Seelig and Granick
Contactdseelig@umn.edu

Project Description: Canine hematopoietic neoplasia is a clinically and biologically diverse set of diseases that are characterized by diseases with uniquely short (e.g., CD34+ acute leukemia; 9 days) and uniquely long (e.g., T-Zone lymphoma; 637 days) survival times. The identification of these unique subtypes often requires a combinational testing approach, including cytology and histopathology. However, many of these unique subtypes, including acute leukemia and T-zone lymphoma, are best diagnosed using flow cytometry. Flow cytometry (FC) is a rapid, objective diagnostic tool that can uniquely provide both diagnostic and prognostic information using samples collected with minimal invasiveness. However, the diagnostic utility of FC is limited by the existence of validated antibodies and tissue-specific reference intervals (i.e., what is considered “normal”). To date, the vast majority of the published literature describing canine FC has documented its utility using either peripheral lymph node or peripheral blood samples. As such, there is limited published data on the flow cytometric findings in canine bone marrow using a modern and diagnostically-relevant panel of antibodies. Owing to the importance of bone marrow flow cytometry for the diagnosis and classification of canine leukemia and in the staging of canine lymphoma, these studies will benefit diagnostic clinical pathologists and veterinary oncologists. The objective of this study is to develop a diagnostic approach for the flow cytometric characterization of canine bone marrow and to generate species-specific reference intervals for nucleated cell populations using tissue-specific and clinically relevant antibodies. These studies will include the validation of antibodies specific to the bone marrow, including CD117 (c-kit) and myeloperoxidase. To do so, we will perform FC on healthy and diseased bone marrow samples from canine patients presenting to the UMN-VMC and UMN-VDL necropsy service. We hypothesize that we will identify unique, tissue-specific RIs that can be used to accurately diagnose hematopoietic neoplasia of the bone marrow.

Techniques Used: Diagnostic and Research applications of flow cytometry,
Bone Marrow Histology and Cytology, Immunohistochemistry

Eligible Funding: Boehringer Ingelheim, NIH T35, Morris Animal Foundation, Dept. Veterinary Clinical Sciences, Skadron Family Scholarship


Project Title: Developing a Novel Cellular Immunotherapy to Treat HIV
Mentor(s): Skinner
Contact: skinn002@umn.edu

Project Description: Nearly 37 million people are infected with HIV. There is no cure or effective vaccine for HIV, and as such there is a need to develop improved therapies. HIV replication is concentrated in B cell follicles, where low levels of CD8 T cells allow ongoing replication. We hypothesize that increasing HIV-specific CD8 T cells in follicles will lead to immune control of infection. We welcome a student to join our team working to engineer virus-specific T cells to home to B cell follicles to functionally cure HIV. We are actively performing pre-clinical studies in animals to test the efficacy of the approach.

Techniques Used: PCR to track therapeutic cells in tissues from treated animals, confocal image analysis of the tissues.

Eligible Funding: Boehringer Ingelheim, NIH T35, Dept. Veterinary & Biomedical Sciences


Project Title: Cancer Immunotherapies: Killing Cancer Cells with Enhanced Leukocytes
Mentor(s): Walcheck
Contact: walch003@umn.edu

Project Description: Human and veterinary oncology face similar challenges, including 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 by leukocyte-mediated cytotoxicity. Our lab is using leukocytes derived from cell lines, induced pluripotent stem cells, and the peripheral blood of humans and dogs to engineer them with enhanced receptors. These receptors are better at recognizing therapeutic antibodies and stimulating the leukocytes to kill cancer cells.

Techniques Used: Techniques the Summer Scholar will be exposed to include cancer cell cytotoxicity assays, leukocyte functional assays, cell culture, and flow cytometry among others. The Summer Scholar will work with trained researchers to learn cutting edge techniques and develop an independent project and novel data.

Eligible Funding: Boehringer Ingelheim, NIH T35, Dept. Veterinary & Biomedical Sciences

Emerging, Zoonotic, and Infectious Diseases

Emerging, Zoonotic, and Infectious Diseases

Project Title: The Role of Dengue Virus NS2B in Pathogenesis
Mentor(s): Aliota
Contact: mtaliota@umn.edu

Project Description: Human dengue virus (DENV) is thought to have originated in nonhuman primates, yet paradoxically human isolates of DENV do not replicate to high titers or generally cause clinical signs in rhesus macaque models. In humans, the DENV-2-encoded protease cleaves STING, reducing type 1 interferon responses and boosting viral titers. Indeed, both sylvatic and human isolates of DENV can universally cleave human STING but are not capable of cleaving nonhuman forms of STING, and this may at least partially explain why dengue disease has been so difficult to model in nonhuman primates. However, it has recently been shown that conversion of residues 78/79 to the human encoded amino acid sequence renders all primate STINGs sensitive to viral cleavage. As a result, this project will experimentally measure type 1 interferon induction in macaque cells by DENV, and use molecular virology and reverse genetic approaches to engineer human DENVs with the ability to cleave nonhuman primate STING. Better understanding the genetics of host tropism will improve upon animal models of dengue disease and provide insight into DENV adaptation and evolution.

Techniques Used: Cell culture, Molecular cloning, Flow cytometry, Classical virology approaches, RT-PCR, and Immune assays

Eligible Funding: NIH T35, Dept. Veterinary & Biomedical Sciences


Project Title: Immune Responses to Viruses Drives Genetic Variation in Swine
Mentor(s): Cheeran and Schroeder
Contact: cheeran@umn.edu

Project Description: Both PRRSV and Influenza A viruses undergo dynamic genomic changes in the swine population making vaccination control program ineffective in preventing viral spread. We hypothesize that the inadequacy of the adaptive immune responses to vaccines, commonly used for control, drive the selection of viral variants that have increased fitness in a new vaccinated host. This project will track the evolution of viral quasi-species generated using in vivo and in vivo experimental models. The goal will be to quantify the pressure imposed by antibody and T cell responses on viral epitopes recognized by pigs. We propose that the unique immune pressures imposed by vaccination with heterologous strains will generate a distinctive viral variant profile.

Techniques Used: Immune assays (measures of humoral and cell-mediated immune responses in pigs), de novo sequencing, evaluation and analysis of viral sequence variation

Eligible Funding: Boehringer Ingelheim, Dept. Veterinary Population Medicine


Project Title: Development of Novel Vaccines Against Bacterial Infection of Swine
Mentor(s): Ly and Liang
Contact: hly@umn.edu

Project DescriptionLawsonia Intracellularis causes proliferative enteropathy in swine. A modified live bacterial vaccine is available, but there are considerable limitations that prevent the widespread use of this vaccine. We plan to develop a new generation of vaccines for L. intracellularis that is based on our novel Pichinde viral vector (PICV), which can overcome some of the known limitations of a modified live bacterial vaccine. Specifically, we plan to generate PICV-based vaccines to express different L. intracellularis proteins and to assess the ability of the monovalent, bivalent, and trivalent vaccines to protect swine from L. intracellularis infection.

Techniques Used: Molecular biology, Virology, Immunology, and Vaccinology
virus and cell culturing, molecular cloning, RNA/protein analyses, animal vaccination, necropsy, histopathology.

Eligible Funding: Boehringer Ingelheim, NIH T35, Dept. Veterinary & Biomedical Sciences


Project Title: Development of New Vaccines to Protect Turkeys Against Hemorrhagic Enteritis Virus Infection
Mentor(s): Ly and Liang
Contact: hly@umn.edu

Project Description: Hemorrhagic enteritis (HE) is an economically important disease of commercial turkeys. The etiologic agent of HE is hemorrhagic enteritis virus (HEV), an avian adenovirus. The vaccines currently available to the commercial poultry producer are effective in preventing disease outbreaks; however, they are immunosuppressive creating opportunities for opportunistic infections and vaccine failures. The main purpose of this project will be to develop and test a new vaccine for HE that are based on a viral vaccine vector (Pichinde virus, PICV) recently developed by our laboratory.

Techniques Used: Molecular biology, virology, immunology, and vaccinology, virus and cell culturing, molecular cloning, RNA/protein analyses, animal vaccination, necropsy, histopathology.

Eligible Funding: Boehringer Ingelheim, NIH T35, Dept. Veterinary & Biomedical Sciences


Project Title: An Investigation into PRRS Viral Evolution using State of the Art Diagnostic Technologies
Mentor(s): Kevill and Schroeder
Contact: jkevill@umn.edu

Project Description: Porcine reproductive and respiratory syndrome (PRRS) viruses are responsible for the largest health-related deaths of U.S swine and cost the swine industry approximately $1 billion each year. PRRS is capable of infecting pigs of all ages and sex. The major disease symptoms include; anorexia, fever, lethargy, respiratory problems, and a range of reproductive issue's such as; late-term abortion, stillborn piglets, and weakened surviving piglets. Vaccinations are often administered and increase herd immunity however, PRRS re-infection and re- emergence are common. The reason why re-infection and re-emergence are common is due to the nature of the PRRS virus, which is a (+)ssRNA virus. (+)ssRNA viruses readily mutate and recombine to form novel viral variants that can evade the host immune response. The aim of this project is to use state of the art technologies to diagnose and determine PRRS viral variation in historic samples, enabling us to see how the virus has evolved over time. The methods used will include RT-PCR and Next Generation Sequencing (NGS). The data generated will provide important information about viral evolution and may explain why PRRS re-infection and re-emergence are common. The student will gain skills at the forefront of molecular biology and genomic data analysis.

Techniques Used: How to work in a lab safely, sample processing, RNA extractions, reverse transcriptase PCR, how to generate sequence data using a MinIon, how to quantify viral samples to establish viral titers, and how to analyze genomic data.

Eligible Funding: Boehringer Ingelheim, Dept. Veterinary Population Medicine


Project Title: Next-Generation Biosurveillance: Rapid Molecular Diagnostics in the Field
Mentor(s): Larsen
Contact: plarsen@umn.edu

Project Description: The miniaturization of genomic technologies allows for the deployment of mobile sequencing laboratories capable of generating vast amounts of DNA and RNA sequence data in remote locations. Such laboratories can be used to facilitate a range of One Health biosurveillance projects aimed at 1) monitoring pathogens circulating in wildlife, livestock, and humans, and 2) providing a molecular barcode of putative host or vector species. To this end, Dr. Larsen has outfitted a mobile sequencing lab based on third-generation nanopore sequencing technology. The Summer Scholars Project will focus on the deployment of miniaturized molecular technology throughout Minnesota in order to test feasibility for rapid on-site molecular diagnostics and to advance One Health biosurveillance initiatives in the State. The project will focus largely on the molecular surveillance of native and invasive species of rodents that are known reservoir species for multiple zoonoses (e.g., hanta virus, Salmonella spp, E. coli, Lyme disease, etc.). Fieldwork will be conducted throughout regions of Minnesota that have been identified as hotspots for emerging zoonoses. The mobile laboratory will be utilized to screen for presence/absence of relevant zoonoses and to sequence specific host genes for molecular identification. Field-based results will be confirmed using traditional molecular approaches (e.g., Sanger sequencing) and by performing phylogenetic analyses. The resulting data will be used to inform ongoing One Health initiatives and will provide a valuable record of pathogen/host genetic variation.

Techniques Used: The student will learn a variety of molecular techniques, including DNA and RNA extraction, PCR, and next-generation sequencing library construction. The student will also learn field-based techniques for the collection and morphological identification of invasive and native species of rodents (mice and rats). These techniques will include habitat selection for live-trapping, setting, baiting, and monitoring live traps, rodent euthanasia, metadata collection, tissue collection, and proper sample-preservation.

Eligible Funding: Boehringer Ingelheim, Dept. Veterinary & Biomedical Sciences


Project Title: Interactions Between Coronaviruses and Aminopeptidase N
Mentor(s): Li
Contact: lifang@umn.edu

Project Description: Aminopeptidase N (APN) is a cellular receptor for many coronaviruses. This project investigates the interactions between coronaviruses and APN using biochemical and cell biology approaches.

Techniques Used: Protein expression and purification, protein biochemistry, cell biology

Eligible Funding: Boehringer Ingelheim, NIH T35, Dept. Veterinary & Biomedical Sciences


Project Title: Reducing Transmission of Bovine Leukemia Virus in Dairy Cattle through Application of Phylodynamics
Mentor(s): Wells
Contact: wells023@umn.edu

Project Description: Bovine leukemia virus (BLV), a retrovirus found on nearly all US dairy operations, causes economic losses through premature culling and reduced milk production of infected cows. It also poses risks to the dairy industry through public health and market access concerns. BLV transmission occurs within herds through blood-borne routes from subclinically infected to susceptible cattle. Though much is known about disease transmission, successful disease control is difficult. We propose to use genotype clustering data available through whole genome sequencing (WGS) of BLV to identify and prioritize routes of transmission using phylodynamic approaches. Samples have previously been collected from an infected dairy herd. We will use WGS for complete genome characterization and to produce SNPs needed for characterization of transmission trees within an infected herd. This will involve integration of WGS and epidemiologic data to evaluate the strength of association with hypothesized transmission pathways within herd (dams to calves through in utero and colostrum/milk, dams to multiple heifer calves born near time of calving through consumption of pooled colostrum or milk, heifer calves to other heifer calves through fomites, biting insects or vaccinations, cows to cows through fomites or biting insects, as well as seasonal risks). Successful completion of this proposed research project will support development of more effective control programs for BLV, part of our longer term goal to reduce between- and within-herd spread of cattle diseases, ultimately reducing financial losses and increasing the competitiveness of US cattle producers.

Techniques Used: Infectious disease epidemiology as relates to bovine leukemia virus (BLV), Inferential epidemiologic analysis, Phylodynamics analysis using WGS data 


Eligible Funding: Boehringer Ingelheim, Dept. Veterinary Population Medicine

Population Systems

Population Systems

Project Title: Use of pre-partum metabolic markers to predict post-partum hyperketonemia
Mentor(s): Caixeta, Godden, Knauer
Contact: lcaixeta@umn.edu

Project Description: The objective of this study will be to develop a multivariable metabolic index (MI) using metabolic markers measured during the pre-partum period to predict the probability of the development of hyperketonemia (KET) during the subsequent lactation in Holstein dairy cows. 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. Although, diet and milk production greatly affect the levels of BHB in circulation during early lactation, a recent studies showed that abnormal pre-partum metabolic and inflammatory responses increases the chances of the development of KET in the post-partum. Thus, in this study, blood samples will be collected from dairy cows at approximately 260 ± 3 days of gestation (21 days prior to estimated calving date) for the measurement of multiple metabolic and inflammatory markers (i.e., total protein, albumin, blood urea nitrogen, calcium, aspartate aminotransferase, alanine aminotransferase, gamma-gluatamyl transpeptidase, non-esterified fatty acids, total bilirubin, BHB, glucose, triglyceride, total cholesterol, sorbitol dehydrogenase) and at days 3±1 and 7±1 for the determination of BHB and diagnosis of KET. We plan to enroll 700 cows in order to identify at least 120 hyperketonemic cows. Animals will be kept under normal conditions at their home herd and will be followed during the first 60 DIM to determine the incidence of diseases. Our results will equip veterinarians and producers to establish optimal herd-level management before the occurrence of KET.

Techniques Used: The student is expected to become proficient in collecting blood as well as develop a better understanding of the management and care of dairy cows around parturition. Additionally, the student working in this project will be responsible for record keeping and analysis of information extracted from on-farm management software.

Eligible Funding: Boehringer Ingelheim, Dept. Veterinary Population Medicine


Project Title: Portable, Multiplex Giant Magnetoresistance (GMR) Biosensor for Simultaneous Detection of Porcine Respiratory Disease Complex (PRDC) Pathogens
Mentor(s): Cheeran and Krishna
Contact: cheeran@umn.edu

Project Description: Porcine respiratory disease complex (PRDC) poses a major challenge to both veterinarians and pork producers. For the pork industry with an annual gross production output of ~$20 billion, respiratory diseases affect up to ~50% of the herd at any given stage of production. There is a need for enhancing the diagnostic capacity for PRDC in production systems. Giant magnetoresistance (GMR) chip along with magnetic nanoparticles (MNPs) have become a powerful tool for highly sensitive and rapid biomolecule detection. The fabrication and integration of GMR biosensors are compatible with multiplexing. GMR-based sensing platforms were found to be matrix insensitive and the absence of ferromagnetism property in biological samples allow detection of a magnetic signal with less background noise. in this project, we propose to develop and validate a rapid, accurate, and affordable test for simultaneous detection of multiple PRDC pathogens using GMR sensors. We have successfully tested a portable handheld GMR platform to detect Influenza virus in swine, with high sensitivity and specificity in the laboratory. In the present project, we will functionalize the GMR biosensor array with the capacity to simultaneous detect influenza virus, PRRSV, PCV2, and Mycoplasma hyopneumonia. This array will be tested using multiple clinical sample matrices to determine its feasibility for use as a pen side assay. This pen-side diagnostic capability could enhance our current detection, surveillance, and control strategies for infectious diseases.

Techniques Used: ELISA, Hybridoma development and selection, Western blots, GMR testing and data analysis

Eligible Funding: Dept. Veterinary Population Medicine


Project Title: Simulating Disease Transmission on Dynamic Contact Networks
Mentor(s): Craft and Gilbertson
Contact: craft004@umn.edu

Project Description: Disease transmission within animal populations is affected by contact frequency and social structure, often represented as contact networks. In wildlife, population density and environmental factors like prey availability and season can influence a population’s connectivity, which may result in temporally dynamic transmission impacts. The goal of this project is to assess how dynamic contact networks in an empirical wildlife system affect disease transmission and epidemic outcomes. This computational project will have implications for wildlife management and conservation.

Techniques Used: The summer student will learn how to perform basic to intermediate coding in the software environment R. The student will learn how to construct contact networks based on empirical data, as well as how to simulate disease transmission on these networks and assess epidemic outcomes.

Eligible Funding: Dept. Veterinary Population Medicine, Morris Animal Foundation


Project Title: The Influence of Land Use and Seasonality on Pathogen Prevalence in Wild Rodents in Minnesota
Mentor(s): Craft, Larsen, and Mistrick
Contact: craft004@umn.edu

Project Description: As the interface between natural and human-dominated environments is increasingly blurred by urbanization, wildlife pathogens of significant concern to human and livestock health are becoming more common. In Minnesota, the mosaic of urban and agricultural areas interspersed with natural forest and grassland habitat facilitates complex interactions among humans, livestock, and wildlife. Wild rodents of the genus Peromyscus are particularly effective at utilizing human-altered landscapes and they thrive in semi-urban environments. Two species of Peromyscus are found throughout Minnesota (P. leucopus and P. maniculatus) and both are natural reservoirs of Giardia intestinalisCryptosporidium parvum, and Borrelia burgdorferi, all pathogens of growing concern for human and livestock health.
The objective of this project is to conduct a pilot study focused on the prevalence of pathogens of human and livestock health concern in wild rodent populations across a range of habitats at the Cedar Creek Ecosystem Science Reserve (CCESR) and the Itasca Biological Station (IBS). Mice will be trapped using a mark-recapture method throughout the summer season and blood and fecal samples will be collected to screen for the presence or absence of G. intestinalisC. parvum, and B. burgdorferi using molecular diagnostics.
We will investigate the questions: How does land use impact pathogen prevalence in wild rodents? Does prevalence change seasonally? Through investigating prevalence across a range of habitats, we hope to gain insight into the processes driving pathogen prevalence in these systems and to better understand the drivers relevant to disease dynamics at the interface of human, animal, and environmental health.

Techniques Used: The student will learn how to conduct research in a field environment to sample for presence/absence of pathogens in wildlife. The student will gain experience handing and processing wild Peromyscus mice, including anesthetization techniques, retro-orbital eye bleeds, and collecting general body measurements. The student will learn how to perform basic coding in the software environment R to conduct statistical analyses on the collected data.

Eligible Funding: Dept. Veterinary Population Medicine, Morris Animal Foundation


Project Title: Multi-Strain Disease Modeling in Swine Production Systems
Mentor(s): Craft and Michalska-Smith
Contact: michalsm@umn.edu

Project Description: Many of the diseases that affect the global swine industry, including influenza, porcine reproductive and respiratory syndrome, and porcine epidemic diarrhea virus, consist of multiple strains or genotypes which co-circulate within the population. These strains might differ in their properties, such as infectiousness, course duration, etc. Moreover, they have the potential to interact through both (partial) shared immunity (an antagonistic relationship) and through an increased risk of co-infection due to immune suppression (a positive relationship). Yet, especially in the complicated system of modern, disseminated pig production, the effects of multiple strains coexisting on overall disease dynamics are poorly understood. Sophisticated models of swine disease dynamics have recently been published, providing a solid foundation for single-strain dynamics. This project seeks to bridge the gap between previous modeling attempts and a multi-strain reality. This project will be primarily theoretical in nature, developing a model of multi-strain disease dynamics on a network of swine farms, but will rely heavily on previously collected data of disease prevalence for validation and parameter fitting.

Techniques Used: Basic programming (in R) and data-management practices.
Foundational disease modeling frameworks, especially compartmental models, and their extensions. Model fitting and model selection procedures.

Eligible Funding: Dept. Veterinary Population Medicine


Project Title: Development of a Sole Ulcer Induction Model in Holstein Cows: the Next Step in Lameness Research
Mentor(s): Cramer, Guedes, and Caixeta
Contact: gcramer@umn.edu

Project Description: Lameness is a pervasive prevalent problem in the dairy industry that affects both animal welfare and farm profitability. A cow with lameness experiences negative changes in her physiology, behavior, and affective state. Sole ulcers are a common, painful cause of lameness. Intervention research on sole ulcers is complicated by their chronic, multifactorial nature, requiring very large-scale longitudinal clinical trials. Our project will address this problem by developing and testing a sole ulcer induction protocol using 48 pregnant Holstein heifers in 2 stages. In the first stage, 24 heifers will be allocated to 6 different groups in a 2-way factorial design, testing the effect of lying time and block application on sole ulcer development. Animals will be enrolled from 60 days pre-calving to 120 days post-calving. Two weeks before calving 4 out of 6 groups of cows will have hoof blocks placed strategically on either 1 or 2 lateral hind hooves to increase pressure under the flexor tuberosity of the 3rd phalanx for 4-6 weeks. Within these groups, cows will be allocated into either restricted lying time (by 6 hours/day) or not. Samples will collected at various time points to describe the developmental process of sole ulcer using various behavioral, metabolic, inflammatory and physical measures. The development of a validated sole ulcer induction model will greatly improve our ability to evaluate sole ulcer intervention strategies.

Techniques Used: Hoof trimming, block application, cattle handling, blood sampling,

Eligible Funding: Dept. Veterinary Population Medicine


Project Title: Validation of the IceRobotics IceQube Tri-axial Accelerometer for Measuring Standing Time, Lying Time, and Steps in Dairy Calves
Mentor(s): Knauer
Contact: knaue020@umn.edu

Project Description: The utilization of technologies to monitor activity are becoming more common on commercial dairy farms and are being used for many management activities including disease detection and reproductive management. While these activity monitors are successfully being used on lactating dairy cattle and breeding age heifers, they are still perhaps underutilized in younger dairy animals. The objective of this study is to validate the IceQube tri-axial accelerometer in dairy calves and heifers to 6 months of age. Data collected from the IceQube (test) will be compared to data collected from video monitoring (gold standard) to determine if the IceQube is a valid measure of standing time, lying time, and step count in growing dairy heifers. The student will conduct this research in a field laboratory setting on a commercial dairy farm in MN, and have the opportunity to present the results of their project at the annual meeting of the American Association of Bovine Practitioners in the fall of 2020.

Techniques Used: With the assistance of the PI, student will be responsible for study design, sampling, data management, analysis, and reporting. The student will gain appreciation for methods used in validity testing, as well as the nuances and challenges of conducting behavioral research in dairy calves and heifers.

Eligible Funding: Dept. Veterinary Population Medicine


Project Title: The Role of the Teat Epithelial Microbiome in Mastitis
Mentor(s): Noyes and Caixeta
Contact: nnoyes@umn.edu

Project Description: This project entails collecting weekly teat skin samples from heifers on various dairy farms in MN. Total DNA from all samples will be extracted and the 16S rRNA gene will be amplified. Amplified DNA will be sequenced and the resulting data will be used to conduct a microbiome analysis of the teat epithelial microbiome. The microbiome data will be compared to mastitis outcomes in the sampled heifers, to try to determine whether certain microbiomes predispose heifers to mastitis.

Techniques Used: Epidemiological study design; on-farm sample collection; use of DairyComp and similar software programs; molecular biological techniques (DNA extraction, PCR amplification, next-generation sequencing library preparation, next-generation sequencing); statistical, bioinformatic and microbiome data analysis; exposure to microbiome research.

Eligible Funding: Boehringer Ingelheim, Dept. Veterinary Population Medicine


Project Title: The Risk of Death: Mortality Rates in Anesthetized Raptors
Mentor(s): Ponder, Allweiler, and Willette
Contact: ponde003@umn.edu

Project Description: In the veterinary literature, birds are often identified as being at a higher risk of mortality associated with anesthesia than other species such as dogs and cats. The few studies and minimal primary literature used to support these statements often have weak case definitions and assumptions that influence the conclusions. At The Raptor Center, we routinely anesthetize multiple species of birds on admission (ill/injured), during recheck exams (increasingly healthy) and during surgery, with associated medical records. Although anecdotal, we believe raptor anesthesia to be as safe as other animals based on our experience of very rare anesthetic-related deaths. We also recognize that the term “bird” reflects many species with many different characteristics.  We propose to contribute to evidence-based decision-making by undertaking a well-controlled retrospective study to look at mortality rates associated with anesthesia in birds seen in wildlife rehabilitation centers. The summer scholar role in this project would be to Undertake a systematic literature review of avian anesthesia, Design and implement a retrospective study on mortality associated with avian/raptor anesthesia utilizing data from The Raptor Center (and possibly other rehabilitation centers), and correlating mortality rates with ASA status, length of general anesthesia (surgery), and signalment (species).

Techniques Used: Literature search/systematic review, development of hypothesis, retrospective study design, data collection/processing, data analysis, scientific writing and communication, clinical handling, avian restraint, anesthetic protocols, anesthetic monitoring (cross-species).

Eligible Funding: Boehringer Ingelheim, Dept. Veterinary Population Medicine, Morris Animal Foundation


Project Title: Identifying Antimicrobial Resistance Determinants of Public Health Concern in Canada Geese
Mentor(s): VanderWaal and Johnson
Contact: millere@umn.edu

Project Description: There is growing evidence that antimicrobial resistant bacteria can spread from anthropogenic sources into natural ecosystems. When these contaminated environmental resources are utilized by humans, there is the potential risk of antimicrobial resistance (AMR) spillback into humans and domestic animals. Canada geese (Branta canadensis) may facilitate AMR spillback given their high densities at environmental sites used by humans (e.g. beaches, parks) and considerable evidence of resistant and pathogenic bacterial strains in goose feces. As such, the overall goal of this project is to characterize AMR in Minnesota populations of Canada geese, with a focus on elucidating the potential public health risks surrounding the presence of AMR-positive geese at the human-environment interface. We will use shotgun metagenomic sequencing to characterize the resistome and associated genetic determinants of mobility and pathogenicity from Canada geese at various geographical locations throughout Minnesota (Aim 1). We will then use the same sequencing data to identify potentially pathogenic strains of Escherichia coli and assess the biological capacity for goose-human transmission (Aim 2). Finally, we will analyze data from Aims 1 and 2 using spatial epidemiology to identify areas of Minnesota with potentially elevated public health risk of AMR spillback (Aim 3). Taken together, this project will improve our understanding of the role wild animals at the human-environmental interface may play in the maintenance and dissemination of clinically-relevant AMR.

Techniques Used: Student’s time will be divided between fecal sample collection from various locations around Minnesota (50%) and laboratory work to process fecal samples for culturing and sequencing (50%). They will participate in determining collection locations and help create the sample collection protocol. Student will learn basic microbiology techniques, including culture and isolation of Enterobacteriaceae from fecal samples and DNA extraction.

Eligible Funding: Dept. Veterinary Population Medicine


Project Title: Health Assessments of Muskrats Translocated to Voyageurs National Park
Mentor(s): Wolf
Contact: wolfx305@umn.edu

Project Description: The National Park Service is translocating muskrats into Voyageurs National Park over a 2-year period to evaluate their effectiveness in the biological control of invasive cattails. A subset of translocated individuals is being implanted with radiotransmitters to evaluate post-translocation movement and survival. As part of that study, we are collecting preliminary data on these individuals to describe overall health and identify potential predictor variables for post-translocation survival. The successful student will assemble and analyze health data sets gathered on muskrats over a 2-yr period. There may also be opportunity for participation in one week of field activities.

Techniques Used: Basic clinical epidemiological analyses and statistics, some wildlife immobilization and handling skills.

Eligible Funding: Morris Animal Foundation, Dept. Veterinary Population Medicine