Summer Scholars Project Proposals

Summer 2022 Project Proposals

Comparative Biosciences

Comparative Biosciences

Project: Molecular Mechanisms of Genome Instability Syndromes
 Dr. Anja Bielinsky ([email protected]), Biochemistry, Molecular Biology & Biophysics

Description:  Our laboratory is focused on understanding diseases that are caused by defects in maintaining a stable genome. These include cancer and cancer predisposition syndromes and congenital diseases caused by mutations in DNA replication and repair genes. Our overall goal is to uncover how a particular mutation alters the function of a protein and how this alteration, in turn, impacts the biological process that this protein is involved in. One recent example is the rare disease of natural killer (NK) cell deficiency caused by the dysfunction of the replication protein MCM10 (minichromosome maintenance protein). Reduction of MCM10 function leads to a premature halt in the development of NK cells but not other lymphoid precursors. Our goal is to explain why this is the case.

Techniques Used:  CRISPR-mediated genome editing and in vitro differentiation of induced pluripotent stem cells to identify the mechanisms underlying the biological alterations caused by specific disease variants.

Project: Protein Phosphatase Phlpp1 in Controlling Bone Loss Associated with High Fat Diet
Dr. Elizabeth Bradley ([email protected]), Dept. Orthopedic Surgery

Description:  High fat diet induces (HFD) many physiological changes, including bone loss in mouse models.  HFD is also associated with declines in bone mass in humans.  Genetic ablation of the protein phosphatase Phlpp1 enhances bone mass accrual in transgenic mouse models.  We also know that Phlpp1 controls responses to inflammatory mediators induced during obesity and in mice placed on HFD.  The goal of this project will to be determine if Phlpp1 protects against HFD-induced bone loss.  Using curated samples, the student would assess bone mass via histology (e.g., sectioning and staining tissues) and perform serum ELISAs for the determination of markers of bone formation/resorption as well as inflammation.  Experiments to determine how Phlpp1 deficient cells respond to inflammatory mediators (e.g., qPCR) will complement these studies.

Techniques Used:  Paraffin sectioning of embedded tissues, histology (e.g., TRAP staining, Masson’s trichrome, immunohistochemistry), serum ELISAs, qPCR.

ProjectRole of Gut microbiome in 2 Models of Hypertension
Dr. John Collister ([email protected]), Dept. Veterinary & Biomedical Sciences              

Description:  Hypertension is the most significant risk factor for myocardial infarction and stroke, the first and third most frequent cause of death in the United States.  New strategies are required to control high blood pressure and in the present project, we will 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 central hypothalamic organum vasculosum of the lamina terminalis (OVLT) in the hypertensive response to both Angiotensin II (AngII) and DOCA (water soluble form of aldosterone) treated rats.  We will examine the intestinal microbiome in two 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 hypertensive rats? 2) Does OVLT lesion restore the microbiome while attenuating hypertension in the rat? 3) Does ileal transplantation from OVLT lesioned rats prevent or treat hypertension?  The following hypothesis will be tested:  OVLT lesion prevents the changes in gut microbiota and increased blood pressure in hypertensive rats. OVLT lesioned instrumented rats will be treated with either AngII or DOCA, and cross-transplanted with ileal content during the hypertensive treatment.  Continuous measurements of blood pressure will be made via radio-telemetry.  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/or placement of the blood pressure transducer catheter in the abdominal aorta.  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 harvesting brain and gut tissues for histological and microbial genomic analyses, respectively.

Project: Resultant Vector Acceleration for Activity Monitoring in Normal Dogs and Dogs with Pain from Chronic Osteoarthritis
Dr. Mike Conzemius ([email protected]), Dept. Veterinary Clinical Sciences              

Description:  Activity monitors (AMs) offer a mechanism to objectively estimate chronic pain in dogs in their natural environment. While many AMs are commercially available, none are validated in the dog and none can undergo quality assurance that would be needed for a regulatory study. We recently completed direct observation validation using resultant vector acceleration in a population of normal dogs performing scripted activities of various intensities. This final stage of validation is to document activity variance in a population of normal dogs and to determine if differences exist compared to a population of dogs with pain secondary to chronic osteoarthritis.

Techniques Used:  Accelerometry, clinical study design, data collection and evaluation.

Project: The Role of Lysosomes in Regulating Drug Resistance in Canine Hemangiosarcoma
Dr. Erin Dickerson ([email protected]), Dept. Veterinary Clinical Sciences              

Description:  Our lab is focuses on the development and translation of new therapies to treat canine hemangiosarcoma. We recently found that lysosomes play a key role in regulating resistance to chemotherapy in hemangiosarcoma cells by sequestering these drugs away from their cellular targets. More resistant cells appear to exhibit enhanced lysosomal biogenesis and increased lysosomal numbers. Our goals are to understand the underlying biology used by cells to regulate lysosome biogenesis and to identify combinatorial approaches to block these pathways. The Summer Scholar will be involved in these novel studies to help advance our understanding of the development of drug resistance in hemangiosarcoma.

Techniques Used:  Cell culture, immunoblotting, cell viability assays, flow cytometry, siRNA transfection, data analysis and presentation.

Project: Understanding the Genetic Basis of Pelger-Huet Anomaly in Australian Shepherd Dogs
Dr. Steven Friedenberg ([email protected]), Dept. Veterinary Clinical Sciences               

Description: Pelger-Huet anomaly is an inherited disorder in Australian shepherds that causes benign abnormalities in the neutrophils and eosinophils of affected dogs. The disease is believed to follow an autosomal dominant inheritance pattern. Dogs with one copy of the mutated gene show classic changes in their white food cells, however dogs with two copies of the inherited mutation are believed to die in utero. The causative gene has been identified in humans, but has never been identified in dogs. In this project, the summer scholar will research the causative gene in humans (lamin B receptor) as well as other genes in the same pathway to develop a list of candidate genes for sequencing. The scholar will then sequence these genes in affected and unaffected Australian Shepherds to identify the causative mutation in dogs. Depending upon these findings, a whole genome sequencing approach may also be employed.

Techniques Used:  Primer design, understanding genetic databases, basic bioinformatics skills, PCR, Sanger sequencing, DNA isolation and extraction, blood smear evaluation, possibly interpretation of next-generation sequencing data.

Project: Developing Immunosuppression-Free Strategies for Transplantation
 Dr. Melanie Graham ([email protected]), Dept. of Surgery, Medical School              

Description:  Our lab has a unique focus on two closely related issues: 1) Developing methods to understand immunometabolism towards innovative therapies for diseases with high public health impact, e.g. diabetes, obesity, and infectious disease, and 2) Identifying the general reasons why animal models don’t always predict human outcomes to improve their accuracy using refinement to elevate animal welfare.  We are developing immunosuppression-free strategies to transplant cells and solid organs using immune tolerance strategies or device-based approaches.  Using islet cells, the insulin producing cells of the pancreas, as an example we have achieved of prolonged diabetes reversal in primates after porcine xenotransplantation, the induction of robust tolerance to allografts, and have recently shown that cells survive for over 6 months using a bioartificial pancreas without any immunosuppression.  Taken together these technologies present a major paradigm shift in transplantation therapy using unlimited tissue sources (xeno- or stem cell- derived).

Techniques Used:  PBMC functional assays; ELISA/RIA; bioenergetics; cell culture; cell and solid organ transplant; flow cytometry; care and monitoring of animals used in research; digital spatial profiling.

Project: Lipid Metabolism and Osteoarthritis
Dr. Alonso Guedes ([email protected]), Dept. Veterinary Clinical Sciences              

Description:  This project focuses on understanding how lipid metabolism contributes to osteoarthritis (OA) pathophysiology. Specifically, our studies seek to determine both the therapeutic efficacy of cyclooxygenase (COX) and soluble epoxide hydrolase (sEH) inhibition in OA and understand the enzymes’ role in OA development/progression. Inhibition of arachidonic acid metabolism with COX inhibitors is first-line therapy for OA in animals and humans. sEH is an enzyme also involved in lipid metabolism that is receiving increasing attention for its role in pain relief and tissue repair. Our overall objectives are i) to elucidate the effect of a COX/sEH dual inhibitor on mobility and cartilage damage in a mouse model of OA and ii) characterize the expression of COX, sEH and related biomarkers in equine OA joint tissues. The proposed research is innovative because it will target the symptomatic and structural aspects of OA and is expected to provide strong evidence-based proof of principle for further development and future clinical trials of dual COX/sEH inhibitors.

Techniques Used:  Mouse models of OA, behavior assessment of pain and mobility, immunohistochemistry, western blots, PCR.

Project: MRI Biomarkers of Canine Intervertebral Disc Degeneration
Dr. Casey Johnson ([email protected]), Dept. Veterinary Clinical Sciences              

Description:  Degenerative disc disease (DDD) is a major health problem for both dogs and humans. Degeneration of the intervertebral discs in the spine can lead to back pain and disc rupture, which can compress and damage the nerves of the spinal cord. There is a clinical need for new, noninvasive imaging techniques to assess the health of discs and predict whether a disc will rupture so that preventative treatment can be applied. Since dogs spontaneously develop DDD, they have significant comparative advantages for studying DDD and its treatment. The goal of this project is to investigate noninvasive, quantitative magnetic resonance imaging (MRI) techniques to detect and characterize disc degeneration in dogs. The summer scholar will compare quantitative MRI measures to the biochemical composition of canine intervertebral discs. A series of canine vertebral column specimens from the thoracolumbar region of the spine will be imaged ex vivo at 3T MRI and then subsequently analyzed with biochemical assays and histology.

Techniques Used:  Quantitative MRI, biochemical assay, and histology data collection and analysis; scientific communication (presentation and writing).

Project: Generating Exogenic Organs and Cells in Chimeric Animals for Regenerative Medicine 
Dr. Walter Low ([email protected]), Stem Cell Institute, Dept. Neurosurgery              

Description:  The transplantation of organs and cells can save lives and reduce disease morbidity.  The severe shortage of donors for human organs, however, has resulted in many patients on waiting lists dying before an organ becomes available.  Recent advances in gene editing and blastocyst complementation have raised the possibility of generating exogenic organs and cells in chimeric host animals as bioincubators for transplantation therapies.  This project addresses the hurdles that currently impede the generation of human organs and cell in animals and its translation into the clinical setting for transplantation in organ and cell replacement.

Techniques Used:  Gene editing with CRISPR/Cas9, tissue culture of human stem cells, embryo manipulation,  immunohistochemistry, microscopy, flow cytometry, animal surgery, RNAseq.

Project: Shine On – Early Detection, Monitoring, and Prevention of Hemangiosarcoma
Dr. Jaime Modiano ([email protected]), Dept. of Veterinary Clinical Sciences                

Description:  We have collected data from dogs with known diagnoses to establish a training set for the Shine On suspicion test (Shine On-phase-1), from dogs with a confirmed diagnosis of hemangiosarcoma that were monitored during treatment to evaluate relapse (Shine On-phase-2), and from healthy dogs older than 6 years of age to be used as a test (validation) set to evaluate risk status for development of hemangiosarcoma or other cancers (Shine On-phase-3). Ongoing work is collecting follow-up data from all the participants in Shine On-phase-3 and retesting a subset of dogs that will be eligible for prevention with the drug eBAT (Shine On-continuation phase).

Techniques Used:  Flow cytometry analysis and applications of statistical methods and models to interpret clinical data.

Project: Animal Models of Pediatric Orthopedic Diseases 
 Dr. Ferenc Toth ([email protected]), Dept. of Veterinary Clinical Sciences              

Description:  The primary focus of the lab is to develop large animal models of human orthopedic disorders. Ongoing projects aim to improve the currently available animal models of Juvenile osteochondritis dissecans (JOCD) and Legg-Calve-Perthes Disease (LCPD). To accomplish these goals the lab evaluates both open and minimally invasive surgical techniques to induce lesions mimicking the human disease then conducts extensive MRI evaluations to follow lesion progression. Final evaluation usually involves histopathology.

The lab also has an emerging line of research involving stem cell mediated repair of focal cartilage defects which provides additional opportunities for training.

Techniques Used:  Surgical procedures, post-operative care of the research animals, MRI and image processing and data evaluation.

Project: Alterations in Mitochondrial Function in Dystrophic Cardiomyopathy 
 Dr. DeWayne Townsend ([email protected]), Dept. Integrative Biology and Physiology              

Description:  Muscular dystrophies are a group of genetic diseases that result in skeletal muscle disease. Many of these diseases also have a significant cardiac disease, which in many cases in the cause of death in these patients. Previous studies have demonstrated that limitations in energetic function may contribute to the pathology of the heart. This project will focus on identifying differences in mitochondrial structure, composition, and function in the hearts of mouse models of muscular dystrophy.

Techniques Used:  Immunohistochemistry, pathological evaluation of cardiac disease, western blotting, and proteomic methods.

Project: Understanding Canine Natural Killer Cells for Killing Cancer Cells
Dr. Bruce Walcheck ([email protected]), Dept. Veterinary & Biomedical Sciences              

Description:  Our lab is actively involved in developing cancer immunotherapies for humans and companion animals. A current focus is on engineering human and canine natural killer (NK) cells with enhanced receptors for better engagement of tumor cells. One of our generated receptors expressed in engineered human NK cells is currently being tested in clinical trials by Fate Therapeutics. We are attempting to apply this same approach in dogs. This involves characterizing canine NK cells, which are not as well understood as they are in humans and mice, as well as designing and testing recombinant receptors and tumor-targeting antibodies. The summer scholar will be involved in these novel studies to help advance our goal of developing effective immunotherapies for cancer in dogs.

Techniques Used:  Tumor cell killing assays, canine leukocyte isolation and characterization, cell culture, and flow cytometry.

Project: Modulating Macrophage Activation to Alter Behavioral Outcomes Following Traumatic Brain Injury
Contact: Dr. Maxim Cheeran ([email protected]), Dept. of Veterinary Population Medicine 

Description:  It is known that traumatic brain injury (TBI), even when it does not result in loss of consciousness, can increase the risks for developing behavioral abnormalities, like cognitive impairment or addiction behavior. The specific mechanisms responsible for these long term behavioral outcomes is currently unknown, although persistent inflammation may have a part to play. The early response to injury in the brain is mediated by infiltrating macrophages. While the macrophage numbers return to baseline after the initial trauma, these macrophages exhibit a proinflammatory activation profile for several weeks, which correlates with appearance of behavioral outcomes associated with TBI. This project investigates the role of these activated macrophages in the development of behavioral dysfunction. Our hypothesis is that classically activated macrophages (i.e. pro inflammatory) exacerbate neuronal dysfunction by remodeling synapses and inhibiting neurogenesis. Using this premise, we are testing ways to modulating the macrophage activation profile in mice to an alternately activated state (anti-inflammatory) to promote neurogenesis and rebuilding of damaged neural circuitry.

Techniques Used:  Flow cytometry, IHC, behavior assays, and RT-PCR.

Project: Neuroinflammation Associated with Sequential Traumatic Brain Injury in a Rodent Model
Dr. Maxim Cheeran ([email protected]), Dept. of Veterinary Population Medicine

Description:  A concussion is the most common type of traumatic brain injury (TBI) and most patients recover without significant CNS pathology. However, these patients are susceptible to the development of neurodegenerative and neuropsychiatric complications after a ‘second hit’ or repetitive TBI, which is common among soldiers and athletes. The present study hypothesizes that a second injury after an initial mild TBI will result in an enhanced neuroinflammatory response, which persists longer and results in greater behavioral deficits. We also posit that treatment with a stem cell therapy will decrease the window of susceptibility for the second hit to elicit an exaggerated immune response in the brain.

Techniques Used:  Flow cytometry, Immunohistochemistry, behavior assays, and RT-PCR.


Infectious Agents

Infectious Agents

Project: Pathophysiology of Emerging Arboviruses
Dr. Matt Aliota ([email protected]), Dept. Veterinary & Biomedical Sciences              

Description:  Zika virus is dangerous to pregnant women and their fetuses/neonates throughout the Americas and beyond. This risk will persist for the foreseeable future, even if it is eventually curtailed by natural immunity and widespread vaccination. Therefore, we strive to provide new tools for pregnant women and their health care providers to assess, and potentially reduce, the risk of congenital Zika syndrome. We combine aspects of immunology, virology, and molecular genetics to understand how viruses like Zika virus emerge and how they subvert host immune defenses. Example topics for summer scholars to develop their proposal around include: (a) molecular evolution of Zika virus during transmission, (b) the role of antiviral signaling at the maternal-fetal interface, or (c) the impact of pre-existing immunity on flavivirus pathogenesis.

Techniques Used:  Cell culture, molecular cloning, flow cytometry classical virology approaches, RT-PCR, and immune assays.

Project: Therapeutic RNA Editing to Halt Viral Pathogens
Contact: Dr. Kathleen Boris-Lawrie ([email protected]), Dept. Veterinary & Biomedical Sciences              

Description:  Coronaviruses and retroviruses are RNA viruses that use host machinery to produce virion proteins. RNA editing is a host process to combat virus proliferation. The molecular basis for RNA editing has been characterized. Two genetically separable domains function for RNA binding to substrate RNA and enzymatic conversion of adenosine to inosine residues in a process known as epigenetic modification. This project will document the RNA editing profile of the 5’-control sequences of avian spleen necrosis virus or human SARS CoV2 fused to reporter RNA. RNA binding domains fused with ADAR1 and APOBEC3G editing domains will be co-expressed in cells with the luciferase reporter RNAs. Editing efficiency and the therapeutic index will be measured using biological replicate assays and biostatistics.

Techniques Used:  Cell culture and transfection, Luciferase assays, DNA and RNA isolation and PCR. Electronic data management. Team science, scientific reading and trouble-shooting.

Project: Screening Natural Products to Halt Virus Propagation in Hosts
Contact: Dr. Kathleen Boris-Lawrie ([email protected]), Dept. Veterinary & Biomedical Sciences              

Description:  Both coronaviruses and retroviruses require host ribosomes to translate viral proteins. To be successful, the viral mRNAs compete with host mRNAs for ribosomes. Viral mRNAs access a specialized translation pathway that is normally reserved for host tumor suppressor mRNAs (JBC 2020). The structural basis for the novel RNA-protein interaction has been defined in detail (NAR 2021, PNAS in press). We are now poised to reverse engineer specialized translation to halt virus propagation. This project will screen natural products in Luciferase reporter assays and produce statistically robust data sets on lead compounds.

Techniques Used:  Cell culture and transfection, Luciferase assays, RNA extraction and RT-qPCR, electronic data management. Team science, scientific reading and communication, trouble-shooting.

Project: Molecular virology of HIV-1 and SARS-CoV-2
Contact: Dr. Alon Herschhorn ([email protected]), Dept. of Medicine        

Description:  We work on exciting research directions to understand and target HIV-1 and SARS-CoV-2 entry at the molecular and cellular levels. We use interdisciplinary approaches at the interface of synthetic and molecular virology, immunology and cell biology to gain new insights into complex biological processes with the aim of translating these insights into novel therapies and vaccines to treat and prevent viral infections. Potential topics include 1. mechanisms of HIV-1 and SARS-CoV-2 resistance to broadly neutralizing antibodies, 2. viral evolution, 3. mapping the vulnerabilities of viral entry, and 4. development of next generation vaccines for viral infections.

Techniques Used:  Molecular biology methods including PCR, cloning, and site directed mutagenesis. Viral neutralization assay. Tissue culture techniques.

Project: Impact of Bacterial Virulence Inhibitors on Transcriptomes of MRSA and Gut Microbiome of Mice
Contact: Dr. Yinduo Ji ([email protected]), Dept. Veterinary & Biomedical Sciences              

Description:  The continuing emergence of multidrug resistant bacterial pathogens has caused great public health concerns due to limited options of antibiotics for treatment of infections. An alternative strategy to combat resistant pathogens is to eliminate their virulence or ability of spread infection. We have identified a novel compound PMI5 that dramatically inhibited the hemolytic activity, invasion, and pathogenicity of MRSA in two mouse models of infection. Moreover, PMI5 had no impact on bacterial growth; however, its derivative PMI4 could inhibit bacterial growth. The reason why the two analogs (PMI4 and PMI5) possess distinct capacity of affecting bacterial growth is totally unclear. The objectives of this project are to determine the effect of PMI4 and PMI5 on transcriptome of MRSA and gut microbiome of mice.

Techniques Used:  All RNA-Seq and DNA-Seq data are generated in UMGC and available. Bioinformatic data analysis, Basic Unix Terminal Commands, RNA-Seq data analysis, microbiome data analysis (QIIME2).

Project: The Impact of R-Loops on Host-Pathogen Interactions
Contact: Dr. Kevin Lang ([email protected]), Dept. Veterinary & Biomedical Sciences              

Description:  R-loops are three stranded RNA:DNA hybrid intermediates that can stall replication forks, cause DNA breaks, and elicit the DNA damage response. Preliminary data from our lab suggest that during infection by bacterial pathogens, human cells must respond to R-loop formation on their chromosomes. This project will map R-loop formation genome-wide using DRIP-Seq (DNA:RNA hybrid Immunoprecipitation coupled with next generation sequencing) in human cells during infection. We will then modulate R-loop levels and measure the outcomes of bacterial infections. The results of this project will help elucidate the connection between host-pathogen interactions and DNA replication in human cells.

Techniques Used:  Tissue culture, bacteriology, bacterial genetics, molecular biology, deep sequencing analysis, genomics, DNA:RNA hybrid immunoprecipitation (DRIP-Seq).

Project: DNA Replication Fork Remodeling During Infection by Bacterial Pathogens
Contact: Dr. Kevin Lang ([email protected]), Dept. Veterinary & Biomedical Sciences              

Description:  DNA replication is an essential function of living cells. Replication is challenged by lesions on DNA, and DNA damage responses must repair these lesions to avoid genomic instability. Several bacterial pathogens generate lesions on DNA during invasion of host cells. The effects these processes have on active DNA replication forks in human cells is not well understood. This project will use iPOND (isolation of Proteins on Nascent DNA) coupled with mass spectrometry to quantify changes in protein composition at remodeled replication forks in living human cells infected with intracellular bacterial pathogens. The results of this project will yield insights into how some of the most common host-pathogen interactions affect core cellular functions.

Techniques Used:  Tissue culture, bacteriology, bacterial genetics, proteomics, iPOND, western blotting.

Project: The Impact of Oxygen Levels on the Mutation Rate of Bacterial Pathogens
Contact: Dr. Kevin Lang ([email protected]), Dept. Veterinary & Biomedical Sciences              

Description:  Mutations are the currency of evolution. The frequency of mutations drives the development of new traits, such as antibiotic resistance. Oxidative stress is a major source of lesions on DNA that lead to mutations. However, the mechanisms of mutagenesis in the absence of oxygen are not well known. This project will leverage novel deep sequencing methods to precisely measure mutation rates in bacterial pathogens grown in the presence and absence of oxygen. We will use Maximum Depth Sequencing, a method that circumvents the error rate of PCR amplification, and other technical artifacts generated during Illumina based sequencing techniques. The results from this project will provide a broader picture of evolutionary pressures experienced by bacterial pathogens.

Techniques Used:  Bacteriology, bacterial genetics, genomics, deep sequencing analysis, Maximum Depth Sequencing.

Project: Comparative Analysis of Retrovirus Replication
Contact: Dr. Lou Mansky ([email protected]), Institute for Molecular Virology                         

Description:  We conduct research on the comparative analysis of retrovirus replication. In particular, we will analyze the evolutionary conservation of key amino acid (a.a) residues for their role in retroviral replication among closely related viruses in the same and other genera of the retrovirus family. Analysis of site-directed mutants for their effects on virus replication will provide important clues for the importance of particular a.a. residues in replication, which may also provide insights for how they may also influence particle infectivity and capacity to be transmitted in nature.

Techniques Used:  Retroviral vector construction; mammalian cell culture; transduction of cells with retroviral vectors; flow cytometry; laser scanning confocal microscopy; site-directed mutagenesis; molecular cloning; nucleotide sequencing analysis.

Project: Discovery of Novel anti-Parasite Drugs for Apicomplexans
Contact: Dr. Roberta O’Connor ([email protected]), Dept. Veterinary & Biomedical Sciences              

Description:  Our lab focuses on discovering drugs with anti-parasitic activity in compounds sourced from marine organisms.  Our main interest is Cryptosporidium, a gastrointestinal parasite of worldwide significance.  However, we also screen against Toxoplasma gondii, as this parasite is easier to manipulate in the laboratory and can serve as a model for the more intractable Cryptosporidium.  The summer student would follow up on one of the hits we obtained from our recent screen of a marine compound library (the HBOI Peak library), that we have shown is effective against Toxoplasma as well as Cryptosporidium. The student would conduct assays to: a. determine the effective concentrations at which 50% and 90% of the Toxoplasma parasites are killed, determine if the compound is effective against extracellular tachyzoites; and characterize the morphology of parasites treated with this compound.  Depending on how quickly progress is made, the student may also investigate the compounds effects on host cell invasion.

Techniques Used:  Cell and parasite culture, immunofluorescence assays, drug susceptibility assays of intracellular and extracellular Toxoplasma parasites (luciferase based), and invasion inhibition assays.

Project: A Portable Magnetic Particle Spectroscopy (MPS) Platform for on-Field Detection of Influenza A Virus and Mycoplasma hyopneumoniae
Dr. Maxim Cheeran ([email protected]), Dept. of Veterinary Population Medicine
Dr. Maria Pieters ([email protected]), Dept. of Veterinary Population Medicine

Description:  Conventional methods for identification of pathogens causing porcine respiratory disease are time consuming and require specialized laboratory facilities. It also involves multiple tests to detect different pathogens. MPS technology can be used to develop an accurate, affordable, fast and easy-to-use test for pathogen detection, including simultaneous detection of multiple pathogens in a single test. We are in the process of developing a hand held MPS system that will enable early, rapid detection of two common porcine respiratory pathogens. In order to develop these handheld tests we have generated monoclonal antibodies (Mabs) to M. hyopneumoniae that do not cross-react with other commensal or other pathogenic species of swine mycoplasma. We will characterize these antibodies for cross-reactivity to M. hyosynoviae and quantify its ability to detect M hyopneumoniae in common clinical sample matrices used for diagnosis of respiratory infection in pigs. We expect that these Mabs when used in a testing system with improved specificity, may provide the swine industry with a more effective tool for screening and monitoring M. hyopneumoniae infection in swine production systems.

Techniques Used:  ELISA development; purification and production of monoclonal antibodies; western blot.

Project:  Immune drivers of PRRSV evolution
Dr. Maxim Cheeran ([email protected]), Dept. of Veterinary Population Medicine
Dr. Declan Schroeder ([email protected]), Dept. of Veterinary Population Medicine              
Dr. Kimberly VanderWaal ([email protected]), Dept. of Veterinary Population Medicine              

Description:  Genetic variation and the presence of multiple co-circulating genetic strains, or "lineages" is the norm for many pathogens, particularly for rapidly evolving viruses, like porcine reproductive and respiratory syndrome virus (PRRSV). Immune response to one lineage may not be completely protective to different lineages. This study will assess the cross-immunity generated by porcine B-lymphocytes to a related lineage. The goal of the project is to elucidate the potential for immune-mediated competition amongst co-circulating lineages and determine how immunity influences evolution of PRRSV.  Using an experimental infection approach, where quasi-species evolution will be tracked via next generation sequencing, the project will explore how immunity-driven factors (humoral immunity), such as vaccination or prior infection, may impose selective pressure on viral evolution and alter the transmission dynamics for multi-lineage pathogens. 

Techniques Used:  B cell assays like ELISA/ELISPOT, virological assays like plaque/focus forming unit assay, and virus neutralization assay, sequencing methodologies: including RNA extraction, sequence analysis methods.

Population Biology and Medicine

Population Biology and Medicine

Project: Disease Ecology Modeling
Contact: Dr. Meggan Craft ([email protected]), Dept. of Ecology, Evolution, and Behavior              

Description:  The student and I will develop a research project on disease transmission within animal populations. The goal of the research would be to understand the spread and control of infectious diseases in wild or domestic animal populations. This would likely involve testing hypotheses regarding infectious disease dynamics by using real world data in mathematical models. Model output can be used to predict disease dynamics and can guide and prioritize future disease monitoring and disease control strategies. In the past, summer students have participated in the design of the study and have capitalized on the opportunity to take the lead on writing a peer-reviewed manuscript.

Techniques Used:  Hypotheses formulation, study design and implementation, data sourcing and cleanup, statistical techniques using the program R, working with a diverse group of collaborators, writing a manuscript (optional).

Project: Improving the Way we Measure the Lameness Status of Cows in the Dairy Industry
Contact: Dr. Gerard Cramer ([email protected]), Dept. of Veterinary Population Medicine              

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. Current locomotion scoring in used to evaluate farms in dairy welfare assurance programs.  In this project, we will correlate those locomotion scores with more readily available hoof lesion data, automated lameness scoring using cameras and digital dermatitis scores. This will require farm visits to locomotion score herds and meetings with hoof trimmers to collect hoof trimming data.

Techniques Used:  Locomotion scoring, hoof trimming/lameness techniques, record analysis, data management.

Project: Reducing Mastitis in the Dairy Cow by Increasing the Prevalence of Beneficial Polymorphisms in Genes Associated with Mastitis Resistance
Contact: Dr. Brian Crooker ([email protected]), Dept. of Animal Sciences                       

Description:  Our premise is that previous selection practices have successfully increased the presence of genetic polymorphisms associated with increased milk yield but have decreased the presence of polymorphisms associated with disease resistance. We will use contemporary Holsteins and unique Holsteins that have not been subjected to selection since 1964 to determine differences in immune and inflammatory responses and mammary gene expression when unselected and contemporary Holsteins are subjected to intramammary challenges with E. coli or S. uberis. A pathogen will be infused into a quarter, and blood and milk samples collected for analysis to assess mastitis severity and the immune response. The genome of the study cows will be sequenced to identify potentially beneficial polymorphic differences between the genotypes. The long-term goal is to increase the presence of polymorphisms in the Holstein cow to strengthen immune function and increase mastitis resistance.

Techniques Used:  Animal handling and care, milking, blood and milk sample collection, and sample processing and analysis including ELISA cytokine assays.

Project: Investigating the Genetic Bases of Cardiac Arrhythmias in Racehorses
Contact: Dr. Sian Durward-Akhurst ([email protected]), Dept. of Veterinary Clinical Sciences              

Description:  Sudden death has a devastating impact on the racing industry and there are major safety concerns. Of the hundreds of horses that die on the track each year, the cause of death is not identified for 47% of them. Cardiac arrhythmias are thought to explain a large number of these cases. Most horses that develop arrhythmias have no underlying structural heart disease, making it virtually impossible to detect those that will develop arrhythmias. Arrhythmias in human athletes without underlying structural heart disease are almost exclusively caused by mutations in ion channel genes. These athletes frequently develop subtle ECG changes that are used for monitoring and to guide when retirement from high level athletic performance is recommended. Given the similarities between these arrhythmias in humans and horses, it is likely that mutations in genes responsible for cardiac arrhythmias in humans are associated with alterations in ECG measurements and the presence of arrhythmias in horses. This project will investigate if mutations in human arrhythmia causing genes are associated with changes in ECG measurements and cardiac arrhythmias in racehorses.

Techniques Used:  Communication with racehorse owners, trainers, jockeys, and grooms, collection and interpretation of ECGs, interpretation of cardiac ultrasound of apparently health racehorses with cardiac arrhythmias, DNA isolation, Agena Bioscience IPLEX bioassay genotyping, and statistical analyses.

Project: Applying a Multiomics Approach to Discovering Risk Factors for Urinary Stone Disease
Contact: Dr. Eva Furrow ([email protected]), Dept. of Veterinary Clinical Sciences                

Description:  Kidney stones are a painful health problem occurring in 9% of the population and accounting for an annual health cost of >$10 billion in the United States. Current therapies to prevent kidney stones have low efficacy, and recurrence rates are high. Like humans, dogs and cats naturally form urinary stones with many shared clinical features of the disease. Our laboratory uses a variety of approaches, including genomics, transcriptomics, metabolomics, and microbiomics, to discover risk factors for urinary stone disease in these natural animal models. The summer scholar will work with the PI to select a discrete aim pertaining to this larger goal, tailored to their interests.

Techniques Used:  Analysis of whole genome sequencing, prioritization of genetic variants, PCR, electrophoresis, DNA sequencing, and analysis of nutritional data in relation to microbiome data.

Project: Systems-Based Approaches in Aquatic Ecosystem Health
Contact: Dr. Amy Kinsley ([email protected]), Dept. of Veterinary Population Medicine              

Description:  Understanding complex interconnected systems at different scales is a difficult but critical component of addressing ecosystem health challenges. Therefore, ecosystem health uses transdisciplinary efforts to bring together scientists, veterinarians, public health practitioners, government and private sectors, and citizens to manage health at the intersection of human, animal, and environmental health. Such transdisciplinary efforts often require systems-based approaches, which take a holistic approach considering how the parts of a system interrelate and fit within the context of larger systems. This project will include a review of scientific literature that will be synthesized and prepared for publication. The study will identify methods for applying systems-based approaches to manage issues relevant to aquatic ecosystem health, such as pathogens, pollutants, and invasive species. The student will have opportunities to assist with other projects in the lab, including field-based and quantitative research focused on natural aquatic systems and aquatic animal health.

Techniques Used:  The student will learn systems-based approaches and ecosystem health; become familiar with common threats to aquatic ecosystem health; learn how to perform a literature search, including data collection, analysis, and visualization; and participate in scientific writing and communication.

Project: Viral Impact on Resistome Dynamics in Swine Fecal Metagenomic Data
Contact: Dr. Noelle Noyes ([email protected]), Dept. of Veterinary Population Medicine              

Description:  Antimicrobial resistance (AMR) poses ongoing public health risks and greatly contributes to economic loss in swine production systems. One way of assessing AMR in populations is through quantification of the antimicrobial resistance genes (ARGs) present in the microbiome, termed the resistome. While traditional AMR studies focus on the direct impact of antimicrobial use on resistome, few focus on how viruses and their associated mobile genetic elements (MGEs) can influence resistome dynamics. The principal goal of this project is to assess the influence of viruses on the fecal resistome of wean to finish swine using time-series metagenomic data. A secondary objective is to recover viral metagenome assembled genomes and compare sequence similarity with resistome patterns over multiple time points.

Techniques Used:  Bioinformatics, metagenome assembly, biostatistics assessment and interpretation, resistome visualization.

Project: ProgRESSVet: Education for Veterinary Services Capacity Building
Contact: Dr. Mary Katherine O’Brien ([email protected]), Center for Animal Health & Food Safety              

Description:  CAHFS is a designated collaborating center for capacity building by the OIE: World Organisation for Animal Health.  In this role, CAHFS designs outreach and education on topics such as risk analysis, global health policy, food safety, biosecurity, and other strategies for controlling infectious disease, particularly at the human/animal interface.  We have current projects in Colombia, Kenya, Uganda, and Vietnam. The ideal candidate is interested in the organizations and policies that support One Health, and someone who wishes to build skills to connect animal health research/sciences to practitioners in the field and the general public. Specific tasks may include reviewing eLearning materials to make recommendations for revision and improvement, finding or creating case studies and examples that enhance the learning for intended audiences, and translating materials for a) cultural appropriateness and/or b) additional languages as appropriate.

Techniques Used:  Creation of learning and outreach materials, best practices for online learning, case study development, systems thinking/One Health approach.

Project: Farm Connectivity and Risk Occurrence in Swine Farms
Contact: Dr. Kimberly VanderWaal ([email protected]), Dept. Veterinary Population Medicine              

Description:  Understanding the spread of pathogens between swine farms is critical for disease control, and different types of farm-to-farm connections may increase the risk of disease occurrence. Traditionally, farms are seen as “connected” if linked via animal movements (animal connectivity) or if located less than a certain distance from one another (spatial connectivity). However, “untraditional" sources of connection between farms may exist and may create opportunities for disease transmission, yet they are rarely quantified. This investigation will measure the association between disease occurrence on swine farms and different types of connectivity between farms.  We will leverage farm status data already collected via the Morrison Swine Health Monitoring Project (MSHMP),  and“untraditional” modes of connection between swine farms will be obtained from a collaborating pig producing company in a region of the US. The risk of disease occurrence (as measured via MSHMP) will be evaluated as a function of different connectivity types between farms. Models evaluating this association will be constructed by the student under mentor’s supervision.

Techniques Used:  The student will gain experience in epidemiology, transmission dynamics, spatial analysis, network analysis, and data visualization. They will be involved in development of statistical models and gain skills in hypothesis formulation, development of epidemiological studies, data collection and processing, analysis and interpretation of study findings.

Project: Generate Global Reference Interval Calculations for Marine Species
Contacts: Dr. Michelle Willette ([email protected]), The Raptor Center       
Dr. Rachel Thompson ([email protected]), Minnesota Zoo              

Description:  Aquariums around the world provide veterinary care for thousands of species, and some of those species have scant reference data available in the literature to aid with interpretation of hematology or chemistry results.  Species360 members can enter this data into their ZIMS electronic medical records and generate aggregated global reference information that is available to veterinary teams around the world.  Several Species360 members have historical records that can greatly augment the dataset for less studied species, but they lack the time to invest in data entry.  The scholar would transform paper or legacy database records into ZIMS for Medical supporting calculation of new reference intervals for pinniped, cetacean, sygnathid, and elasmobranch species. References for these species may not be available in any other aggregated form, and it will continue to grow as more data is added to ZIMS in the future, creating a dynamic and growing resource for aquatic veterinary medicine.

Techniques Used:  Best practices for data entry of medical test results into ZIMS, evaluation and comparison of test methodologies, calculation routines for reference intervals as defined by the American Society of Veterinary Clinical Pathology, introductions to Species360 Conservation Science Alliance team members and data management practices.  This is an international group who impact wildlife trade and conservation science worldwide and partner with groups such as IUCN,CITES, and the Max Planck Institute.

Project: Development and Evaluation of Chronic Wasting Disease Biosecurity Practices using a Risk Assessment Approach by Farmed Cervid Producers
Contact: Dr. Scott Wells ([email protected]), Dept. of Veterinary Population Medicine               

Description:  Chronic wasting disease (CWD) poses an existential threat to cervid farms due to continued disease spread.  CWD preventive practices are focused on postmortem surveillance and preventing transmission through regulatory policies and implementation of biosecurity practices.  However, transmission is possible through multiple exposure pathways to susceptible farms.  This project will address the critical need to develop and evaluate on-farm educational information and tools to facilitate adoption of biosecurity practices on cervid farms to reduce risks of CWD introduction. This will involve creation of web-based interactive CWD biosecurity educational learning modules including an on-farm risk exposure tool and evaluation of the impact of implementing biosecurity program on cervid operations that implement the biosecurity program.

Techniques Used:  Infectious disease epidemiology as relates to chronic wasting disease, development of biosecurity educational materials for use by cervid farms, analysis of study farm data to assess willingness to implement specific biosecurity practices to reduce risk of CWD transmission, communication with project team members, government agency representatives, and cervid farmers.

Project: Serological Patterns of Disease in Moose
Contact: Dr. Tiffany Wolf ([email protected]), Dept. of Veterinary Population Medicine                     

Description:  Serological disease screening can provide important insights into individual or population health. Since 2010 serological disease screening has been conducted on moose at time of capture and/or mortality in northeastern Minnesota on the Grand Portage Indian Reservation.  Moose are an important subsistence species for Grand Portage Band of Lake Superior Chippewa, yet the population has been declining.  We have captured and obtained serological disease data from 147 individual adult moose over 10 years.  We plan to investigate spatiotemporal patterns of disease, including examining relationships between disease and habitat use, survival and reproductive success.

Techniques Used:  R programming skills, reproducible research, data cleaning, correlation analysis, logistic regression, survival analysis, spatial analysis.