vet.ucalgary.ca · Web viewThese results, suggest that stirred suspension bioreactor processing could be used to generate large numbers of SKPs in a relatively short period of time

2016 UCVMResearch Day

April 1, 20168:00am – 3:00pm

Theatre 4 & HRIC Atrium

Registration (Theatre 4): 8:00am

Session 1: 8:15am – 9:15amChinju JohnsonGuilherme Rizzoto

2016 UCVM Research Day

Mina OjaghiAlysha Dance

Session 2: 9:30am – 10:30am

Paula Valenzuela-LeonAlysson Macedo da SilvaSepideh AbbasiWisoo Shin

Session 3: 10:45am – 12:15pm

Shruti SrivastavaSrinivasarao KarriChimone DaltonKrishna BhattAdam ChernickMarie Worobets

Lunch: 12:15pm – 1:00pmPoster Viewing (HRIC Atrium): 1:00pm – 3:00pm

Jessica Davis-Unger

Justin DuvalSarah EngbersCarrie FischerJami Frederick Persephone Greco-OttoAndrew HagnerShubha JainSrinivasarao KarriAlessa KuczewskiAmy LarkinVivian LeungMarta MarchiniMelissa MoggyEmily MorabitoSangay RinchenMonica Rincon GarciaMaria Alejandra Santa Wisoo ShinRobyn ShipclarkHolly SparksSimrika ThapaEmily Zhang

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PLATFORM PRESENTATIONS

Elevated testicular temperature alters semen parameters and sperm DNA integrity in bullsChinju Johnson, Ariana Dirk, John Kastelic and Jacob Thundathil

1. Department of Production Animal Health, Faculty of Veterinary Medicine, University of Calgary

The objective was to determine effects of elevated testicular temperature on morphology, motility, and DNA integrity of bull sperm. Ejaculates were collected from sexually mature Holstein bulls (n=3) twice weekly for 10 consecutive weeks. Ejaculates collected during Week 1 served as controls, whereas testicular temperature was elevated by scrotal insulation for 72 consecutive hours during Week 2. Semen samples were evaluated for sperm morphology and motility. In addition, DNA integrity of frozen-thawed sperm from these ejaculates was determined by sperm chromatin structure assay (SCSA). Elevated testicular temperature substantially decreased sperm parameters (motility and normal sperm morphology), and there was significant negative correlations between these sperm parameters and proportion of sperm with altered chromatin integrity (motility; r= -0.78; normal morphology: r= -0.86). We inferred that DNA of germ cells at various stages of development in the testis was more susceptible to elevated testicular temperature compared to that of epididymal sperm. In conclusion, exposure of testicular sperm to elevated temperature impaired spermatogenesis and sperm function in bulls. The interval from increased testicular temperature to appearance of compromised sperm in the ejaculate was dependent on its developmental stage at the time of increased temperature.

Effects of cryopreservation on content and activity of sperm Na/K-ATPase in beef bullsGuilherme Rizzoto, Gayathri Rajamanickam, Ariana Dirk, John Kastelic and Jacob Thundathil


We demonstrated that both content and activity of testis-specific isoform of Na/K-ATPase (ATP1A4) were significantly correlated to field fertility of dairy bulls. Using a cohort of mature beef bulls (n = 9) classified as satisfactory potential breeders, the objectives of this study were to: (i) compare content (flow cytometry and immunoblotting) and activity (colorimetric assay) ATP1A4 in fresh sperm; and (ii) evaluate effects of cryopreservation on content and activity of ATP1A4. Results demonstrated that sperm content of ATP1A4 differed among beef bulls. In addition, content of this protein was significantly correlated with its enzyme activity. Although cryopreservation procedures did not alter ATP1A4 content, it significantly reduced its activity in frozen-thawed sperm versus fresh sperm from the same ejaculates. In summary, beef bulls classified as satisfactory breeders differed in sperm ATP1A4 content and activity. Moreover, semen cryopreservation procedures altered ATP1A4 activity without altering ATP1A4 content. Although the link between ATP1A4 and field fertility of beef bulls remains currently unknown, ATP1A4 may serve as a fertility marker to better predict variations in field fertility of bulls classified as satisfactory potential breeders.

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Testes-specific isoform of Angiotensin Converting Enzyme (tACE) is involved in regulation of bovine sperm capacitationMina Ojaghi, John Kastelic and Jacob Thundathil


Bulls classified as satisfactory breeders based on traditional breeding soundness evaluation (BBSE) differ in fertility. Therefore, additional tests are necessary to complement BBSE to better predict field fertility of bulls. Since DNA of mature sperm is transcriptionally inactive, sperm functions are regulated by proteins present in mature sperm. Therefore, the overall objective was to investigate role of tACE in regulation of bull sperm capacitation. Specific objectives were to: 1) develop a tACE antibody specific for cattle; 2) characterize expression of tACE in bovine testicular tissue and sperm; and 3) determine the role of tACE in regulation of sperm capacitation. Our custom anti-tACE antibody detected tACE (110 kDa) from sperm protein extracts and immunlocalized this protein in the sperm head and principal piece region of the sperm tail. We demonstrated that tACE shedding occurred during bovine sperm capacitation and tACE activity in the capacitation medium was required for successful capacitation of bovine sperm. Future studies will be focused on evaluating a potential association between field fertility of bulls and tACE content in their sperm.

Nutritional modulation of reproductive potential in dairy bullsAlysha Dance, John Kastelic and Jacob Thundathil


Holstein bull calves often reach AI centers in suboptimal body condition. Early-life nutrition is reported to increase reproductive performance in beef bulls. The general objective was to determine effects of early-life nutrition on reproductive performance of Holstein bulls. We hypothesized that Holstein bull calves on high-nutrition diets during early life will have larger testes, greater sperm production, and be younger at puberty than those on low-nutrition diets. We determined that bulls fed the high-nutrition diet from 2-31 wk were younger at puberty, had larger testes and had higher concentrations of circulating IGF-I than low-nutrition bulls. Furthermore, high-nutrition bulls had more proliferating and differentiating Sertoli cells earlier in life than low-nutrition bulls and IGF-I promoted proliferation of bovine Sertoli cells in culture. Overall, we demonstrated that IGF-I had an important role in bovine reproduction and we identified key periods during bull development where it appeared that nutritional modulation was affecting testes development. Therefore, nutritional modulation can be recommended as a management tool to improve reproductive potential of AI bulls. In addition, we inferred that nutritional modulation may serve as a model for studying mechanisms controlling reproductive development in bulls.

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Autophagy in porcine germ cellsPaula Valenzuela-Leon and Ina Dobrinski

1. Department of Comparative Biology & Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary, Calgary, Canada

2. CONACyT (National Council of Science and Technology), Government of Mexico

Autophagy is a conserved cellular process by which cells sequester a portion of their cytoplasm and organelles into double-membraned vesicles called autophagosomes. Upon autophagosome maturation, the outer membrane fuses with the lysosome membrane causing degradation of the contents as well as the inner membrane by lysosomal enzymes to generate amino acids and other cellular building blocks for recycling by the cell. Autophagy also functions as quality control mechanism for proteins and organelles as it targets and degrades misfolded proteins or impaired organelles. The autophagic pathway is induced as a result of different stress conditions, such as starvation, hypoxia, and DNA damage. Knowledge of autophagy is extensive in somatic cell physiology, though its role in germ cell physiology is much less understood. Therefore, the objective of this study is to investigate the presence of autophagy in porcine germ cells. Testis tissue from 1 and 10 week old pigs and primary cells obtained from a 10 week old pig testis by enzymatic digestion and were analyzed for the presence of autophagy using antibodies for p62 and LC3 via immunostaining and Western blotting. The results showed that the autophagic pathway is not activated in germ cells in the testis and is activated only after the cells are obtained. Cells were then plated for 24 hours under different culture conditions, namely with and without serum, and in StemPro medium. Germ cells still undergo autophagy in all conditions, though in StemPro medium germ cells had fewer autophagosomes and a lower level of autophagy was confirmed in the Western blots. Maintaining porcine germ cells in culture for long periods of time has so far been largely unsuccessful. The use of autophagy as a marker for viability and overall germ cell wellbeing could aid in defining culture conditions more representative of the situation in vivo that may be required to maintaining these cells for a longer time in culture.

The Implications of Estrogen in Maternal Recognition of Pregnancy in HorsesMacedo, A.; Klein, C.

1. Department of Veterinary Clinical and Diagnostic Sciences, Faculty of Veterinary Medicine

The equine conceptus is surrounded by a thin but tough glycoprotein capsule between days 7 and 21 of gestation. Because of the capsule, the equine conceptus does not elongate as described in other species, keeping the characteristic spherical shape of the equine embryo. Moreover, the equine conceptus shows a continuous, self-induced mobility throughout the uterine lumen between days 6 and 16 after ovulation to spread the unknown maternal recognition of pregnancy factor throughout the uterine surface; consequently, ensuring a close communication between conceptus and endometrium. During this phase, the equine embryo can be anywhere in the uterine lumen; nevertheless, at the moment of first detection (Days 9-11 after ovulation), it is often found in the uterine body. The embryo migrates throughout the uterine lumen from one horn to another 10 to 20 times a day until it ceases mobility at around Day 16 after ovulation. Throughout preimplantation phase, numerous possible signaling molecules are secreted into the uterus by the conceptus, for example PGE2, alpha-hydroxyprogesterone and estrogens. Thus far, studies performed with estrogens were based on exposing cycling mares to estradiol instead of eliminating estrogen production by the conceptus; which may address remarkable information with regards maintenance of the corpus luteum.

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Investigating the fate of dermal stem cells in severe skin injuriesSepideh Abbasi and Jeff Biernaskie

1. Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine

It has previously shown that when the skin excisions are larger than 1 cm, de novo hair follicles (HFs) form in the middle of the wound bed after re-epithelization. This wound-induced hair follicle neogenesis parallels the development of HFs during embryogenesis. It has been demonstrated that the nascent HFs arise from epidermal cells outside HFs and not from the bulge stem cells within the pre-existing HFs. However, the origin of the mesenchymal compartments of these neogenic HFs is still unclear. We hypothesized that the mesenchymal compartments of the de novo HFs originate from aSMA+ (alpha-smooth muscle actin) population of the cells within the dermis. To explore this, αSMA-CreERT2-ROSAYFP mice received Tamoxifen at postnatal day 3/4 to label dermal stem cells (DSCs) and their progeny. Full-thickness excisions (>1cm) were made on the dorsal skin of the mice. The wounds are harvested at different time-points after wounding to assess the mesenchymal cells of the neogenic HFs. Preliminary findings show that aSMA+ DSCs contribute to wound healing, generating interfollicular dermal fibroblasts and repopulating newly formed HFs after severe skin injury. They also appear to re-assume their role as DSCs within the de novo HFs and maintain their integration even after three hair cycles.

Hair Follicle Dermal Stem cells (DSCs) show reduction in self-renewal capacity and terminally differentiate to exit the stem cell niche with increasing ageWisoo Shin, Sarthak Sinha and Jeff Biernaskie

1. Department of Comparative Biology and Experimental Medicine

Dermal stem cells (DSCs) reside within the hair follicle (HF) mesenchyme and function to repopulate the dermal papilla (DP) and dermal sheath (DS) with each HF regenerative cycle. With increasing age, the mice skin exhibits a significant reduction in HF density and delayed hair growth following injury. Based on this, we asked whether this age-dependent decline in HF regeneration is due to impaired DSC function. When we performed long-term fate mapping of DSCs using αSMACreERT2:ROSAYFP mice over 18 months, we observed a 22% decline in the number of YFP+ telogen HFs in aged skin. This suggests there is either a progressive loss of DSCs or the DSCs are regularly replaced. However, we did not observe any differences in the number of YFP+ DSCs per telogen HF in young versus aged mice suggesting the size of the stem cell pool remains the same in aged skin. In performing clonal fate mapping of individual DSCs in young versus aged skin, we found that aged DSCs show reduced capacity to self-renew and terminally differentiate more frequently, exiting the HF stem cell niche. Understanding the impact of age on DSCs may enable stem cell-based strategies to rejuvenate skin function following aging, injury and disease.

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In-silico investigation of parasite proteomes to predict immune mimicking proteinsShruti Srivastava 1 , Derek M. McKay2, and James D. Wasmuth1

1. Faculty of Veterinary Medicine, University of Calgary2. Cumming School of Medicine, University of Calgary

Helminths, parasitic worms, are major cause of mortality and morbidity in humans and livestock around the world. Infections are typically chronic, enabled by parasite induced changes in the immune responses, which includes a reduced inflammatory response. Proteomic analyses in several helminth species have implicated parasite-derived proteins as modulators of the host immune system, but their identity is unknown. Understanding the molecular components of this host-parasite interaction offers potential new treatments for severe auto-immune diseases, including: inflammatory bowel diseases, autoimmune encephalomyelitis, and arthritis. Here, we have taken a genomic-based approach to identify short peptides which potentially mimic host proteins, particularly components of the immune system. Our focus has been on the tapeworm species: Echinococcus, Taenia and Hymenolepis. To overcome the problems typical with eukaryote genomes (e.g. fragmented assemblies and missing annotations), we have searched the published gene models, transcriptomes and DNA sequence reads. To date, we have identified 35 peptides that are potential mimics, of which ten mimic proteins involved in the host immune system. These are undergoing structural modelling, the results of which will be presented.

Increasing the cellular levels of hydroxycholesterol interferes with PrPSc propagation in neuronal cellsSrinivasarao Karri and Sabine Gilch

1. Department of Ecosystem and Public Health, Faculty of Veterinary Medicine, University of Calgary

Prion diseases are transmissible, fatal neurodegenerative diseases of mammals, and are characterised by accumulation of isoform of cellular prion protein (PrPC), known as PrPSc (‘Sc’ for Scrapie) in the brain. Today, no curative therapy is available for these disorders. Recent findings published by our laboratory demonstrated that cholesterogenic gene expression is up-regulated in prion-infected compared to non-infected neurons. Other data suggests that interference with cholesterol metabolism inhibits PrPSc propagation. All these findings demonstrate the importance of host cholesterol metabolism for prion propagation. Our data in prion infected mouse neuroblastoma cells (22LN2a5), neuronal cells (22LCAD5, RML CAD5) show that down-regulation of cholesterol synthesis or increasing the levels of hydroxyl-cholesterol by using cholesterol analogues or modulators results in reduction of PrPSc propagation. Additionally, we have identified an orally available drug (Efavirenz), which activates cholesterol-24 hydroxylase and thereby reduces brain cholesterol levels, crosses the blood-brain-barrier, reduces brain cholesterol levels and strongly interferes with PrPSc conversion in different prion-infected cell lines. We will validate this effect in mouse bioassays in order to prove the therapeutic potential of increasing levels of 24-hydroxy-cholesterol in vivo.

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Exploring herpes virus as a tool to study wild animalsChimone Dalton and Frank van der Meer

1. Department of Ecosystem and Public Health

Nearly every animal is infected with at least one, if no many, species-endemic herpes virus (HV). HVs are double-stranded-DNA viruses that co-evolve with their natural mammalian hosts. Transmission of HV occurs directly and frequently in a population causing life-long, often asymptomatic infection. When compared to whole host genomes, HVs contain fewer genes making genomes shorter and simpler. Variation in HV genomes therefore become evident and less prone to confounding factors as seen with host mitochondrial DNA. HVs such as Herpes simplex virus 1 (HSV-1) have shown evidence to support the “Out-of-Africa” hypothesis of historical human migration. HVs have been implicated in wildlife, yet no one to date has explored these viruses to study historical animal evolution. We will focus on these wildlife-endemic HVs to study the evolutionary history of animals in Canada. HV will be examined as a phylogeographic tool in herbivores (Rangifer tarandus; caribou) and carnivores (Martes americana; marten). We hypothesize there is a strong correlation between the relatedness of endemic HV sequences, and the natural geographic distribution of its animal host. HV phylogeography may yet reveal new evidence relating to existing theories of animal invasion and colonization while reducing the dependence on complex host genomics or archeological evidence.

Characterization of antimicrobial and metal resistance in contemporary (2012-2015) and historic (1980s) clinical isolates of Histophilus somni from feedlot cattle in AlbertaKrishna Bhatt 1 , Edouard Timsit1, Andrew Potter2, Claire Windeyer1, Karen Liljebjelke1

1. Faculty of Veterinary Medicine, University of Calgary, Calgary, AB, Canada2. Vaccine and Infectious Disease Organization, University of Saskatchewan, SK, Canada

Histophilus somni is one of the causative agents of bovine respiratory disease and also causes other systemic infections such as myocarditis, pericarditis, pneumonia, pleuritis, arthritis, and thrombotic meningo-encephalitis. The purpose of this study is to determine antimicrobial and metal resistance in contemporary (2012-2016) and historic (1980-1990s) clinical isolates of H. somni. Antimicrobial and metal susceptibility was determined by broth micro-dilution and agar dilution method, respectively. Ninety-six percent of contemporary isolates (n = 63) were resistant to at least one of the antimicrobials tested, and 62 % were multidrug resistant. A significantly (p<0.0001) higher percentage of resistance was observed in contemporary isolates versus historic isolates against: gentamicin, neomycin, oxytetracycline, penicillin, and tylosin. Historic isolates were sensitive to all other classes of antimicrobials, except gentamicin and neomycin. Minimum inhibitory concentration (MIC90) for oxytetracycline, penicillin, tylosin, and chlortetracycline was increased by 32, 16, 8, and 4 times, respectively, in contemporary isolates versus historic isolates. Contemporary isolates had MIC values higher than 1 mM (MIC of reference strain), for Cu (64%) and Zn (59%). Results of this study will be useful in making effective treatment and management decisions in feedlots and will help reduce feedlot mortalities, production losses and risk of developing antimicrobial resistance.

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Bovine Viral Diarrhea Virus Quasispecies Structure in Persistent Infection Adam Chernick and Frank van der Meer


Bovine viral diarrhea virus (BVDV) is a ssRNA virus infecting cattle. Although most economic losses result from transient infections, persistent infections (PIs) are essential to maintaining ongoing transmission. PIs result from viral transmission from dam to fetus in utero early in gestation. These animals are immunotolerant to the virus and remain infected for life. Their high levels of viral shedding expose other cattle in the herd to BVDV. The nature of infection and lack of immune responses to BVDV in PI animals results in a viral quasispecies. We have studied this in a collection of PI animals using next generation sequencing on viral RNA obtained from mesenteric lymph nodes. We have developed a method of constructing and comparing codon- and amino acid-level variant frequency profiles. In this way we have shown that the overall profile of variability is generally conserved between PIs. The structural genes are more variable than the non-structural with a few important exceptions. Additionally, there are animal- and strain-specific differences in the amount of variability at some points in the genome. These results help us to better understand the evolution of BVDV in PI animals which are essential to the cycle of infection on a farm.

In-utero and post-mortem magnetic resonance imaging of ethanol-exposed fetal macaque brainM.L. Worobets, X. Wang, C.D. Kroenke, & C. Rolian

1. UCVM2. Oregon National Primate Research Center, Oregon Health & Sciences University, Division of

Neuroscience, Advanced Imaging Research Center (OHSU)

Prenatal maternal alcohol consumption is a leading cause of environmental related birth defects and mental retardation in the western world. Women who consume ethanol prior to maternal recognition of pregnancy represent a significant number of fetal alcohol spectrum disorder (FASDs) cases. Current detection of FASDs occurs only in mid- to late childhood when cognitive and behavioural symptoms first appear. Identifying new anatomical biomarkers will facilitate earlier diagnostics and treatment strategies. Non-invasive in-vivo magnetic resonance imaging (MRI) is used to characterize teratogenic effects of prenatal ethanol-exposure in human and animal models where decreases in cerebral cortical volume are observed. In-vivo MRI studies of cerebral cortical maturation must be carried out in-utero because the relevant period of development occurs prenatally in primates. We performed in-utero and postmortem fetal brain MRI procedures on macaque dams exposed to 1.5g/kg/day of ethanol or calorie-matched quantities of maltose/dextrin throughout the first 60 days of gestation. Fetal cerebral cortical surface areas at gestational day 110 or 135 were investigated using manual segmentation. Preliminary results suggest that smaller cerebral cortical surface areas are detectable in ethanol-exposed fetuses using in-utero MRI procedures. In-utero MRI shows promise as a prenatal diagnostic tool able to characterize the effects of maternal alcohol consumption.

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NOTES

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POSTER PRESENTATIONS

Risk factors associated with beef cattle lameness within southern Alberta feedlotsJessica Davis-Unger, Karen-Schwartzkopf-Genswein, Sonia Marti, Ed Pajor, and Karin Orsel

1. Department of Animal Health, Faculty of Veterinary Medicine, University of Calgary

Lameness within Alberta feedlots is a significant animal welfare concern. Lameness causes negative impacts to animal production, welfare such as pain and discomfort and also economics. Risk factors such as age, gender, breed, weather, season, pen size, pen condition, days on feed, and ration may impact the risk of lameness development. Understanding the relationships between risk factors and lameness will give producers valuable information to manage this disorder. Data was collected from 2005 - 2015 from 20 southern Alberta feedlots. The producer-collected data was done using the Medlogic program. Analysis will consist of multiple-step wise logistic regression models to determine the relationships between the risk factors and lameness. Our results can clarify what the important risk factors are, explaining the occurrence of lameness in Alberta feedlots. These results can show producers the implications of risk factors and lameness within the feedlot. It will aid producers in making changes to their management systems such as preventative or mitigation strategies. Strategies changes can improve animal welfare such as mitigating pain and discomfort caused from lameness. Understanding the magnitude of lameness within Alberta feedlots can help prioritize lameness as a concern of beef welfare in the beef industry.

The cardiopulmonary effects of dexmedetomidine, alfaxalone, and isoflurane combination anesthesia in pigsJustin Duval, Nigel Caulkett, Jessica Paterson, Soren Boysen

Objective: Determine the cardiopulmonary effects of partial intravenous anesthesia (PIVA) using constant rate infusions (CRI) of dexmedetomidine and alfaxalone with inhalant isoflurane. Study design: Prospective, non-randomized, non-blinded, using eight castrated male Landrace-White swine. Methods: Dexmedetomidine, alfaxalone, and azaperone was administered intramuscularly. Alfaxalone was used for induction. CRI’s of dexmedetomidine, and alfaxalone, with isoflurane and oxygen were used for maintenance anaesthesia. Respiratory rate (RR), heart rate (HR), systemic and pulmonary arterial pressure, central venous pressure, cardiac output, bispectral index, cardiac index, systemic vascular resistance, oxygen delivery, oxygen extraction ration, oxygen consumption, and arterial and mixed venous blood gases were recorded every 10-minutes for 60 minutes after titrating the expired isoflurane concentration to 1.1-1.5. Results: There was a significant increase in RR (P=0.0118), minute volume index (P=0.0405), VO2I/DO2I (ER) (P=0.0412), arterial diastolic pressure (P=0.0486), and significant decreases in PvO2 (P=0.0003), SvO2 (0.0457), hemoglobin (P=0.0104), CaO2 (P=0.0141), CvO2 (P=0.0028), pulmonary diastolic pressure (P=0.0248), and HR (P=0.0413) over time. Conclusions and clinical relevance: The few cardiopulmonary changes noted with this PIVA protocol were within clinically acceptable ranges for swine. This anesthetic protocol may be a suitable option for swine where the cardiovascular and pulmonary systems are of particular interest.

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Evaluation of a supra-glottic airway device for provision of general anesthesia in rabbitsSarah Engbers 1 , Amy Larkin1, Mahesh Jonnalagadda2, Melanie Prebble3, Nic Rousset3, Cameron Knight4, Daniel Pang1

1. Veterinary Clinical and Diagnostic Sciences, UCVM2. Animal Resource Centre, University of Calgary3. Western Veterinary Specialist and Emergency Centre4. Department of Ecosystem and Public Health, UCVM

Introduction: Rabbits have a high incidence of anesthetic-related death compared to other companion animals (Brodbelt et al., 2008). This is influenced by a variety of factors including difficulty in obtaining a secure airway (Eatwell, 2013). Methods: 13 healthy New Zealand white rabbits were enrolled in a block randomized trial comparing endotracheal intubation (ET) to a novel supraglottic airway device (V-gel) during general anesthesia. Computed tomography (CT) scans were conducted before and after insertion of each airway device and the rabbits were monitored (capnography, arterial blood pressure) throughout. After one hour of anesthesia, the rabbits were euthanised and necropsies performed. Results: Both devices successfully maintained general anesthesia. The tip of 6/7 V-gels protruded into the larynx, reducing cross-sectional area of the airway, though this area did not differ from the ET group. Airways were better sealed in the ET group. End-tidal CO2 and arterial blood pressure did not differ between groups.Significance: V-gel position within the oropharynx is sub-optimal, but an adequate airway is provided.

Evaluating and managing pain associated with surgical castration in young piglets using ParacetamolCarrie D. Fischer, Jessica Law, Egan Brockhoff, Joseph M. Stookey, and Claudia Klein

1. Faculty of Veterinary Medicine, University of Calgary, AB2. Prairie Swine Health Services, Red Deer, AB3. Large Animal Clinical Sciences, University of Saskatchewan

The pain experienced by food-producing animals during routine on-farm procedures, such as castration, is a growing welfare concern. A recent study described a novel model for assessing pain in piglets following castration, using navigation times through a linear chute system containing hurdles. Paracetamol (acetaminophen) is a therapeutic agent with analgesic and anti-pyretic actions. Its ability to mitigate pain associated with castration in piglets is unclear. The aims of this study were to use navigation time as a tool for assessing pain and to evaluate the analgesic effects of paracetamol. Navigation times in 2 to 5 day old male piglets were measured prior to, at, and following surgical castration. Paracetamol (60 mg) was administered orally at the time of castration (t=0h) or 15 minutes prior. Navigation times of untreated, castrated piglets were greater than uncastrated control piglets at t=0h. Navigation times of paracetamol-treated piglets were not significantly different from untreated castrates at any time point. There were no significant differences between uncastrated controls, sham-treated castrates, and castrates pre-treated with paracetamol. Results from this study suggest that navigation times alone may be useful for measuring pain levels immediately at castration, but not for post-operative pain assessments. The analgesic effects of paracetamol remain unclear.

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The effects of oral Yunnan Baiyao on blood coagulation in beagles measured using kaolin-activated thromboelastography and buccal mucosal bleeding timesJami Frederick, Søren Boysen, DVM, DACVECC, Catherine Wagg, DVM, DACVP (Clinical Pathology), and Serge Chalhoub, DVM, DACVIM (SAIM)

1. Faculty of Veterinary Medicine, University of Calgary

Objective: To examine effects of oral Yunnan Baiyao (YB) on hemostasis by measuring coagulation via buccal mucosal bleeding times (BMBT), and citrated kaolin-activated whole blood thromboelastography (TEG). Design: Randomized controlled crossover trial. Animals: 8 University owned beagles. Procedures: Beagles were given either placebo or 1000mg Yunnan Baiyao orally every 12 hours for 5 consecutive treatments. Baseline blood was drawn 24 hours before treatment. Blood was drawn again 2 and 24 hours after the last treatment. BMBTs were performed at each time point in duplicate. TEG analysis was performed in duplicate 60±5 minutes after sample collection. Results: There were no significant differences between control and treatment group BMBTs or TEG parameters at any time points. Significant differences were found between baseline and 24 hour time points within the treatment group for LY30 and LY60, and between baseline and 24 hour post-treatment time points within the control group for MA, G, LY30 and LY60 values. No adverse effects were noted. Conclusion: Oral YB, at the dose and frequency administered in this study, did not appear to have any clinically significant effects on measured coagulation parameters. Within treatment group differences were likely due to analytical error and not administration of YB as similar differences were seen in the control group. Further studies with a larger sample size, as well as direct measures of platelet function, are needed.

Physiological effects of pulling competition in heavy horsesPersephone Greco-Otto HBSc; Shannon Massie BScKin, CEP, MSc; Erin Shields HBSc, DVM; Edmond Pajor BSc, MSc, PhD; Renaud Léguillette DVM, PhD

1. Department of Veterinary Clinical and Diagnostic Sciences, Faculty of Veterinary Medicine, University of Calgary, Calgary, AB T2N 4N1, Canada. (Greco-Otto, Massie, Shields, Pajor, Léguillette).

Background: The Heavy Horse Pull is a competition where teams of two horses pull an increasingly heavy sled. Similar to wrestlers and other athletes, many of these horses undergo rapid weight loss in order to enter at a lower weight category. The prevalence and physiological effects of this practice are currently unknown in horses. Methods: 50 horses were divided into light, middle and heavyweight categories based on their arrival weights and then compete 1-3 days after. Bodyweight was measured upon arrival and pre-competition. Blood was sampled at arrival, pre- and post-competition.Results: Bodyweight increased significantly between arrival and pre-competition for all weight categories, with an increase of 7.2% (35.64±25.16 kg), 8.6% (69.10±26.00 kg) and 0.8% (12.70±15.22 kg) for light, middle and heavyweight categories, respectively. Change in body weight was correlated (r2=0.79, p=0.004) with ranking for middleweight horses. The ratios of weight pulled to team bodyweight were 2.7±0.3, 2.7±0.1 and 2.4±0.3 for the lightweights, middleweights and heavyweights, respectively. Blood chemistry indicated moderate to severe hemoconcentration in 16/34 horses on arrival, 11 of which were middleweights. Muscle enzymes were elevated on arrival in 6/34 horses and 11/20 horses after the competition. Conclusions: This study was the first to show the effect of pulling competition on physiological parameters in draft horses. The observed change in weight negatively affected the physiological parameters of many of the horses, but did not appear to impair performance.

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Gene expression profiling within the hair follicle dermal stem cell lineageAndrew Hagner, Whitney Alpaugh, Matt Workentine, Eko Raharjo, and Jeff Biernaskie

1. Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary

Within each hair follicle (HF), specialized mesenchymal cells provide instructive signals to epithelial cells to maintain continuous tissue regeneration. We recently demonstrated the existence of a self-renewing dermal stem cell (DSC) that functions to populate both mesenchymal compartments within the HF: the inductive dermal papilla (DP) and connective tissue sheath (CTS), which wraps around the transient portion of the follicle. In order to identify: 1) mesenchyme-derived activators of HF regeneration, and 2) DSC-lineage determinants, we developed a novel FACS strategy to isolate DSCs or their differentiated progeny (DP and CTS) from early growth follicles, then performed RNA-seq to identify gene expression signatures for each population. Signature genes were validated using qPCR and immunohistochemistry. Ongoing work is examining the functional role of certain candidate genes within hfDSCs and HF regeneration, in part using a conditional knockout strategy and topical application of candidate agonists. Together, our findings impart novel insights on the molecular regulators of the hfDSC lineage and the potential activation signals of epithelial progenitors at the onset of HF regeneration.

Cell Culture-based Models for studying Chronic Wasting Disease PrionsShubha Jain, Sampson Law, Sandi Nishikawa, Yuzuru Taguchi, Jeff Biernaskie, and Hermann Schätzl

1. Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary

Chronic wasting disease (CWD) of cervids is the most contagious prion disease. CWD prion infectivity is found in many tissues, body fluids and secretions. Cell culture models provide the opportunity to study the cellular and molecular biology of prion infections. Mostly scrapie-derived and mouse-adapted prions have been studied in cell models. Currently, only modified RK13 cells are available as persistently infected model for CWD prions. We want to develop new cell culture models for acute and persistent propagation of CWD prions. Our rational is to use cell lines derived from cervid tissue which endogenously express cervid PrPc. We have established skin-derived stem cells from antler tissue of Caribou as such stem cells. In addition, we use a non-transformed fibroblast cell line from Indian Muntjac deer, as fibroblasts are known to be well susceptible to prion infection. Using immunoblot and confocal microscopy analysis we found that both cell lines express normally glycosylated PrPc and that PrPc is located at the outer leaflet of the plasma membrane. We are now in the process of infecting these cells with CWD prions derived from brains of transgenic mice or deer and elk. PrPSc uptake and prion propagation is studied using immunoblot and confocal microscopy read-outs. Novel CWD prion propagating cells will be useful for elucidating the molecular mechanisms underlying the pronounced lateral spread of CWD prions.

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Increasing the cellular levels of hydroxycholesterol interferes with PrPSc propagation in neuronal cellsSrinivasarao Karri and Sabine Gilch


Prion diseases are transmissible, fatal neurodegenerative diseases of mammals, and are characterised by accumulation of isoform of cellular prion protein (PrPC), known as PrPSc (‘Sc’ for Scrapie) in the brain. Today, no curative therapy is available for these disorders. Recent findings published by our laboratory demonstrated that cholesterogenic gene expression is up-regulated in prion-infected compared to non-infected neurons. Other data suggests that interference with cholesterol metabolism inhibits PrPSc propagation. All these findings demonstrate the importance of host cholesterol metabolism for prion propagation. Our data in prion infected mouse neuroblastoma cells (22LN2a5), neuronal cells (22LCAD5, RML CAD5) show that down-regulation of cholesterol synthesis or increasing the levels of hydroxyl-cholesterol by using cholesterol analogues or modulators results in reduction of PrPSc propagation. Additionally, we have identified an orally available drug (Efavirenz), which activates cholesterol-24 hydroxylase and thereby reduces brain cholesterol levels, crosses the blood-brain-barrier, reduces brain cholesterol levels and strongly interferes with PrPSc conversion in different prion-infected cell lines. We will validate this effect in mouse bioassays in order to prove the therapeutic potential of increasing levels of 24-hydroxy-cholesterol in vivo.

The Development of a Bovine Leukemia Virus Control ProgramAlessa Kuczewski and Frank van der Meer

1. Supervisor: Frank van der Meer2. Department of Ecosystem and Public Health

Bovine Leukemia Virus (BLV) infects cattle and affects mainly dairy herds. It is present on 90% of all Albertan dairy herds. Besides the development of tumors in about 5% of infected animals, the virus disrupts the immune system of cattle, resulting in an impaired immune response. By causing a production limiting disease, the infection with BLV results in financial losses to producers and industry.The development of a producer-driven and developed Bovine Leukemia Virus Control Program for Alberta intends to raise awareness and reduce the prevalence of virus and disease. The combination of communication and knowledge exchange with a variety of stakeholders, a small implementation-trial and on-farm workshops will result in the creation of a high-quality, practical and sustainable control-program. After an individual risk assessment a specific action plan will be proposed that can be tailored to individual needs. An important part of this risk assessment is the detection of infected animals. The use of the testkit that was found to be the best among commercially available testing platforms will make this possible in a reliable way. To underline the necessity of a control program an economic assessment of the current situation in Alberta and Canada will be conducted.

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Comparison of the Rat Grimace Scale and a composite behavioural pain scale in ratsCassandra Klune, Amy Larkin, and Daniel Pang

1. University of Calgary Faculty of Veterinary Medicine2. Hotchkiss Brain Institute3. Markin4. NSERC

Currently, there are two spontaneous pain scales validated for rats; the Rat Grimace Scale (RGS) and a composite behavioural pain scale (CBPS). The RGS scores facial expression whereas the CBPS measures frequencies of postural changes (e.g. back arching, staggering, and writhing). These scales have never been directly compared. Our aim was to compare the scales in a post-operative pain model in rats. Female rats (n = 45) were randomized to receive saline, the non-steroidal anti-inflammatory drug meloxicam (2 mg/kg) or the opioid buprenorphine (0.05 mg/kg) 30 minutes prior to laparotomy. Rats were video-recorded 30 minutes pre-operatively and 30, 150, 270, and 390 minutes post-operatively. Videos were blinded and assessed using both scales. The saline treatment group showed a significant increase in pain scores post-operatively with both the RGS and the CBPS with peak pain scores occurred at 270 minutes post-operatively. CBPS scores did not increase following treatment with meloxicam or buprenorphine, indicating an analgesic effect. In contrast, RGS scores in rats treated with meloxicam increased significantly post-operatively and did not differ significantly from the saline control group, whereas pain scores remained low following buprenorphine treatment. These data indicate that the pain scales studied are non-equivalent.

Cage-side pain assessment in real time with the Rat Grimace ScaleVivian Leung and Dr. Daniel Pang

1. Department of Veterinary Clinical and Diagnostic Sciences, Faculty of Veterinary Medicine, University of Calgary, AB, Canada

The Rat Grimace Scale (RGS) is a validated pain scale for assessing spontaneous pain behaviour in rats via facial expressions. Facial expressions are scored using 4 action units: orbital tightening, nose/cheek flattening, ear and whisker changes. Each action unit is scored from 0-2, where 0 is normal (no pain) and 2 is abnormal (pain). Currently, scoring is performed on images from video recordings. While effective, the standard method is laborious and limits timely intervention, as scores are assigned hours-days later. This study evaluated usage of the RGS in real time (RT) to assess pain in an inflammatory pain model. Male and female Sprague Dawley rats (208-402g) were randomised to receive a subcutaneous injection of either buprenorphine (0.03 mg/kg) or saline (12/group), followed by intra-plantar 0.1% carrageenan 30 minutes later. Observations were made at baseline, 3, 6, and 9h post-injection. Observation periods lasted 10 minutes, with an observer performing RT scoring (every 30s). Concurrent video-recording for offline image capture (every 3 minutes) and scoring (IMG) was performed for comparison. Videos and images were randomised and blinded. Data from this study indicate that RT scoring is effective and feasible, potentially allowing the RGS to be applied for cage-side scoring.

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Investigation of an uncharacterized mutation in mice with osteochondrodysplasia-like phenotypeMarta Marchini 1 , Marek Kucka2, William H. Beluch2, Yingguang Frank Chan2, Campbell Rolian1

1. Faculty of Veterinary Medicine University of Calgary2. Friedrich Miescher Laboratory of the Max Planck Society, Tuebingen, Germany

Osteochondrodysplasia is a disorder that disrupts normal bone and cartilage growth and often leads to a short limb phenotype. Mutations in several genes are known to be implicated in osteochondrodysplasia, including Fgfr3, Sox9, and Gli3. Many are autosomal dominant and if recessive have low postnatal viability. Our lab has established a line of mice with an uncharacterized autosomal recessive mutation that exhibits an osteochondrodysplasia-like phenotype. Using microCT data and histology, we have begun to characterize the phenotype of the recessive mice. These mice are characterized by a ~30% length reduction of the tibia and ulna relative to control CD1 mice. An extra thoracic vertebra is variably present in homozygous recessives and heterozygotes. Histologically, the long bone growth plate in recessive mice shows no substantive differences when compared with wild-type. However, microCT and histology data suggest there is less trabecular bone, and it is more disorganized. This combination of gross skeletal and histological abnormalities has not been reported in any known osteochondrodysplasia mouse model. Preliminary genetic data suggest the mutation is localized to chromosome 1, and may involve a cell adhesion gene. Developmental genetic analyses are currently underway to further characterize this mutation.

A mixed methods study of management practices associated with cattle pain and stress in Western Canadian cow-calf operationsMelissa Moggy¹, Edmond Pajor¹, Wilfreda Thurston¹, Sarah Parker², Angela Greter³, Karen Schwartzkopf-Genswein⁴, John Campbell², Claire Windeyer¹

1. University of Calgary¹2. University of Saskatchewan²3. Alberta Farm Animal Care³4. Agriculture and Agri-Food Canada⁴

Public concern for animal welfare is driving change in the agricultural industry. The Canadian “Code of practice for the care and handling of beef cattle” was revised to reflect this change. Implementation of the Code is dependent on its feasibility and importance to producers. However, there is a lack of information regarding current management practices. The objective of this study was to describe pain and stress-associated practices implemented on farm and producer perceptions towards mitigation strategies. The quantitative aspect of this study involved a questionnaire delivered to 109 cow-calf producers in Western Canada. The qualitative aspect involved 15 respondents sampled based on questionnaire responses for interviews. The prevalence of mitigation strategies reported for dystocia and caesarean section were 54.8% and 100%, respectively. The majority of operations did not utilize pain mitigation for castration, dehorning, and branding (88.2%, 94.2%, and 3.9%, respectively). Thematic content analysis was used to explore themes concerning participant rationale behind the implementation of pain and stress mitigation practices. Themes included: access to information and resources, age of the animal, benefit to operation, cost and logistics, market demands, and personal conscience. The results of this study may provide direction for future policy-making, research, and extension efforts.

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Cow comfort, evaluated by lameness, leg injuries, and lying time, after facility and management changes in dairy barns in Alberta, CanadaEmily Morabito, Herman W. Barkema, Ed Pajor, Laura Solano, and Karin Orsel


Cow comfort is a key component to maintaining animal welfare, decreasing lameness, and decreasing injuries. In 2011, 81 freestall dairy barns in Alberta, Canada, were evaluated on animal based measures and environmental measures. The results of these evaluations were benchmarked, allowing producers to assess their farm’s cow comfort in regards to the Canadian Dairy Code of Practice, as well as farm averages across Canada. The purpose the current reevaluation trial (2015) was to determine what farmers have changed, and to quantify the impact of these changes. Fifteen farms were selected that made changes to their freestalls between 2011 and 2015, and were compared to 15 farms made changes to areas not related to the freestalls between 2011 and 2015. Average hours of lying time, as well as prevalence of lameness, knee injuries, and hock injuries in farms that made changes changed from 10.800.65(h/d), 20.333.96, 8.564.02, and 6.832.41% to 11.481.24(h/d), 11.071.29, 5.562.25, and4.021.96%, respectively (p<0.05). On farms that did not make changes, there were no differences between 2011 and 2015. Farms that made changes to the freestalls improved in cow comfort and animal welfare when compared to farms that made changes not related to the freestall area.

Qualitative risk assessment for the re-introduction of rabies into rabies free parts of BhutanSangay Rinchen and Susan Cork

1. University of Calgary, Department of Ecosystem and Public health, Faculty of Veterinary Medicine

Rabies is a fatal viral zoonosis responsible for thousands of human deaths and massive economic losses globally. Bhutan is a small Himalayan country where 70% of the population (est.700,000) depend primarily on Agriculture. Rabies was once endemic across Bhutan but through rigorous mass sterilization and vaccination of dogs, Northern parts of the country have not reported a human case for almost two decades. Southern parts still remain endemic. In the south there are about 16 outbreaks in dogs annually with spill over to cattle. This results in considerable economic losses as a result of livestock deaths and human post exposure treatment. Sixteen human deaths have been reported since 2006. Control of rabies in the south remains difficult because Bhutan shares its entire southern border with India where the disease is highly endemic and animal movement across the border is hard to regulate. Bhutan aims to eliminate rabies by 2020 and is currently working to declare the northern regions rabies free. However, with enhanced trade, growing complexity in the human-dog relationship, and more animal transportations across Bhutan, the risk of re-introducing rabies to the Northern regions remains high. A qualitative risk assessment was undertaken to identify possible pathways of rabies introduction and to identify disease mitigation options.

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Patterns of diversity of bovine viral communities (bovine virome) in fecesMonica Rincon Garcia and Frank van der Meer


Viruses are the most abundant biological entities on earth. They play key roles in the shaping of microbial communities in different environments and hosts. Studies of viral abundance and diversity will lead to novel insights into the functioning of gut microbiota in animals. We processed 4 bovine feces samples to generate metagenomic studies to compare diversity from sick and healthy cattle. The samples are being processed using virus particle purification methods which include filters with different pore sizes, gradient centrifugation and nuclease treatments prior to nucleic acid extraction. The use of a combination of signature gene characterization and next generation sequence will allow us to characterize and compare fecal viro-diversity by these two different procedures. The signature gene PCRs will discover the diversity of viruses starting with the Myoviridae, Herpesviridae and Reoviridae families and the gene Rotavirus. The NGS will allow us to compare the diversity by richness and abundance of known species and discover novel genes and viruses present in the samples. Non-parametric viral discovery curves will be used to assess the total number of viruses. A two-mode affiliation network will illustrate the connectivity between viruses, and finally phylogenetic trees will infer the viral composition of the feces samples.

The distribution of the European strain of Echinoccocus multilocularis in AlbertaMaria Alejandra Santa-Sepúlveda

1. Department of Biological Sciences, Faculty of Sciences2. Department of Ecosystem and public health, Faculty of Veterinary Medicine

Echinoccocus multilocularis is a zoonotic cestode considered as a global emerging pathogen. Recent findings indicate that the parasite is expanding its range and the European strain is circulating in Western Canada in intermediate and definitive host. This represents a high public health risk due to this strain is deemed to be more pathogenic than North American strain and there is evidence of its present in urban areas in Alberta. The aim of this study is to assess the spatial distribution of the European strain of E. multilocularis in coyote and fox populations in Alberta, the genetic variability of the parasite and its differences with the European clades to determine the time of invasion and to estimate the minimum number of invasions events that have occurred in Alberta, identifying possible sources of invasions. We expect to find a wide distribution of the parasite with higher prevalence of the European strain and different European genotypes with a relatively low genetic distance from the original European clades that would confirm that the invasions occurred in recent times. This will help to a better understanding of the epidemiology and ecology of the parasite that allow the assessing of zoonotic risk transmission.

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Hair Follicle Dermal Stem cells (DSCs) show reduction in self-renewal capacity and terminally differentiate to exit the stem cell niche with increasing ageWisoo Shin, Sarthak Sinha and Jeff Biernaskie

1. Department of Comparative Biology and Experimental Medicine

Dermal stem cells (DSCs) reside within the hair follicle (HF) mesenchyme and function to repopulate the dermal papilla (DP) and dermal sheath (DS) with each HF regenerative cycle. With increasing age, the mice skin exhibits a significant reduction in HF density and delayed hair growth following injury. Based on this, we asked whether this age-dependent decline in HF regeneration is due to impaired DSC function. When we performed long-term fate mapping of DSCs using αSMACreERT2:ROSAYFP mice over 18 months, we observed a 22% decline in the number of YFP+ telogen HFs in aged skin. This suggests there is either a progressive loss of DSCs or the DSCs are regularly replaced. However, we did not observe any differences in the number of YFP+ DSCs per telogen HF in young versus aged mice suggesting the size of the stem cell pool remains the same in aged skin. In performing clonal fate mapping of individual DSCs in young versus aged skin, we found that aged DSCs show reduced capacity to self-renew and terminally differentiate more frequently, exiting the HF stem cell niche. Understanding the impact of age on DSCs may enable stem cell-based strategies to rejuvenate skin function following aging, injury and disease.

Pattern of Thyroxin Induced Transformation in Axolotls (Ambystoma mexicanum)Robyn Shipclark B.Sc. and Jason Anderson B.Sc., Ph.D.

1. Faculty of Veterinary Medicine, University of Calgary, Alberta, Canada

The axolotl (Ambystoma mexicanum) is a salamander that never metamorphoses to a terrestrial adult stage, reaching sexual maturity in their aquatic larval stage (a condition called neoteny). Metamorphosis can be experimentally induced in axolotls with thyroxin, but the exact details of the cranial morphology of the transformed adult have not been previously described. To determine how cranial morphology changes across, and compares to other ambystomatids after, metamorphosis, we use 3D digital data from high resolution CT before treatment with thyroxin and the 4 weeks following treatment. Forcing the transformation resulted in regression of vomers, coronoids and palatopterygoids and expanded of ossification of bones such as the squamosals and quadrates. By contrast staining the soft tissues we added cranial muscles, nerves and blood vessels to the 3D models. Expansion of the interpterygoid vacuity and location of the retractor bulbi is similar to adult anurans which retract their eyes to help them swallow prey, and may represent a previously-underappreciated lissamphibian (or batrachian) apomorphy. Future studies using geometric morphometrics and kinematics can be used to quantify feeding specializations such as gape and compare them to patterns seen in the fossil record. The knowledge of the progression of tissue remodeling during metamorphosis can be applied to study regenerative medicine and developmental biology.

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Skin derived precursor cells (SKPs) expanded in computer controlled stirred suspension bioreactors maintain de novo hair follicle inductive capabilities in vivoBreanna S. Borys, Natacha A. Agabalyan, Holly D. Sparks, Kathryn Boon, Sepideh Abbasi, Jeff Biernaskie, Michael S. Kallos

1. Schulich School of Engineering, University of Calgary2. Faculty of Veterinary Medicine, University of Calgary

One major obstacle facing all exogenous stem cell-based therapies is in developing a safe and efficient bioprocess that is capable of generating sufficient numbers of cells, whilst maintaining their inherent functional properties and preventing transformation. Our previous work has demonstrated the existence of a dermal stem cell in adult mammalian skin, which can be isolated and expanded in vitro (called ‘skin derived precursor cells’ or SKPs). When transplanted to full thickness skin wounds, SKPs are ‐able to generate new dermal tissue (Biernaskie et al, 2009). Thus SKPs may represent a renewable source of cells to regenerate dermal tissue to improve skin wound healing. Here we asked whether expansion of adult rodent SKPs in stirred-suspension bioreactors is able to maintain their inherent functional properties. Adult GFP rat SKPs were expanded in computer-controlled stirred suspension bioreactors which provided a 2-fold increase in multiplication ratio relative to static culture. Equal numbers of SKPs from each condition were then transplanted in combination with epithelial aggregates in a de novo hair follicle induction assay or into normal skin of NOD SCID mice to assess homing ability and potential tumor formation. SKPs exposed to bioreactors exhibited similar capacity to induce new follicle formation and repopulation of hair follicles compared to SKPs grown in static culture. Phenotypic characterization is in progress. These results, suggest that stirred suspension bioreactor processing could be used to generate large numbers of SKPs in a relatively short period of time that maintain clinical functionality.

Modulation of protein quality control pathways as a novel intervention strategy in prion diseasesSimrika Thapa, Manel Ben Aissa, Basant Abdulrahman and Hermann Schätzl

1. Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary, Calgary, Alberta, Canada

Increasing numbers of neurodegenerative disorders pose tremendous disease and economic burden on our aging society. Prion diseases are prototypical protein misfolding diseases in human and animals caused by misfolding of the cellular prion protein (PrPC) into the infectious PrPSc isoform. These diseases can be transmitted between or within species, including zoonotic transmission. It is therefore critical to understand the molecular and cellular mechanisms of prion propagation and transmission to elucidate molecular strategies for disease control. Previous studies have demonstrated a direct influence of impairment in quality control mechanisms on prion conversion. In this study, we want to manipulate cellular quality control pathways in vitro and in vivo by stably over-expressing selected folding and sorting proteins which will represent a novel pathway counteracting prion propagation. We are using a cutting-edge lentiviral gene therapy technique to stably transfer therapeutic target genes into prion-infected cells and animal models and determine its effect on prion conversion. The candidate targets are proteins involved in proteasomal degradation (EDEM1, EDEM2 and EDEM3), folding (ERp57) and secretory cargo transport (ERGIC-53 and VIP36). The overall outcome of this work will provide new mechanistic insights into prion infections and define a novel therapeutic strategy against prion diseases and protein misfolding disorders.

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The influence of rater training on reliability and accuracy of the Rat Grimace ScaleEmily Zhang and Dr. Daniel Pang

1. Faculty of Veterinary Medicine, University of Calgary

The Rat Grimace Scale (RGS) was developed to score facial expressions from still images. Inter-observer reliability for the RGS was reported by the scale developers, but the role of observer training has not been evaluated. We aimed to assess the impact of training on inter-observer reliability. Images scored by an expert observer were used for structured training of 4 observers with no experience with the RGS. Multi-step training: 1. 42 images (S1) scored independently with the aid of a training manual, followed by group review with the expert rater. 2. New set of 150 images (S2a) scored independently. 3. Following group review, the same image set (S2b) was re-scored independently. Inter-observer reliability was assessed with an intra-class correlation coefficient (ICC) and results classified as: ‘‘very good’’ (0.81–1.0), ‘‘good’’ (0.61–0.80), ‘‘moderate’’ (0.41–0.60), ‘‘fair’’ (0.21–0.40), ‘‘poor’’ (< 0.20) (Altman, 1991). Group ICC scores (95% CI) improved from “moderate” for S1 (0.58 [0.43-0.72]), to “good” for S2a (0.68 [0.58-0.76]), with a significant increase to “very good” for S2b (0.85 [0.81-0.88], p<0.05). These data suggest an important training effect and highlights the critical role of training in successfully adopting this pain assessment tool.

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The Office of Graduate Education would like to extend our many thanks to the following people who helped to make our 2016 UCVM Research Day a success:

UCVM Faculty & Graduate Students

Distinctive CateringIT Department, Medicine

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All VMS Faculty and StaffThe Office of Graduate Education

Thank you!

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Documents

vet.ucalgary.ca · Web viewThese results, suggest that stirred suspension bioreactor processing could be used to generate large numbers of SKPs in a relatively short period of time