Veterinary Clinical Sciences - Theses

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    Isocitrate dehydrogenase 1 R132H: epidermal growth factor receptor and Ki-67 expression in canine gliomas
    Fraser, Anne Rosemary ( 2019)
    The diagnosis, histological classification and grading of canine gliomas can be challenging, particularly in the antemortem. In humans, it is essential to accurately diagnose gliomas as the treatment and prognosis vary between tumour histologic type and grade. Numerous biomarkers have been investigated to aid in the diagnosis and classification of human gliomas, including epidermal growth factor receptor (EGFR), Ki-67 and a point mutation in the isocitrate dehydrogenase 1 gene at codon 132 (IDH1 R132H). Only limited studies looking at these biomarkers have been performed in dogs, from which clear conclusion cannot be drawn. The prognosis for canine glial tumours is considered guarded to poor, with the current treatment options having limited success. Consequently, there is a lack of gold standard treatment for canine glioma. In humans, gliomas are being investigated at a molecular level to identify therapeutic targets and to improve responsiveness to treatment. Numerous molecular targets have been identified including EGFR and IDH1 R132H. Canine gliomas share many morphologic, histologic and immunohistochemical characteristics with human gliomas, suggesting a translational approach to the classification, treatment and prognostication of canine gliomas may be possible. The objectives of this research were: (1) to describe the signalment, presenting clinical signs, diagnostic findings, treatment and survival time of a series of dogs with glial tumours, (2) to investigate the series of canine gliomas for the IDH1 R132H mutation, and for EGFR and Ki-67 expression, utilising immunohistochemistry and (3) to determine if immunoreactivity is associated with tumour histologic type and grade. The clinical data of thirty-one dogs with histologically confirmed glial tumours was reviewed retrospectively. The formalin-fixed paraffin-embedded tumour specimens were evaluated for IDH1 R132H, pan-IDH1 (IDH1 wild-type and mutated IDH1), EGFR and Ki-67 expression. IDH1 R132H and pan-IDH1 expression were recorded as positive or negative. EGFR immunoreactivity was evaluated using a semi-quantitative score, and Ki-67 expression was expressed as the Ki-67 labelling index (LI). Patient signalment, clinical presenting signs and diagnostic investigation findings were similar to previous reports. The IDH1 R132H point mutation was not identified in any (0%) of the canine glial tumours, while all 31 (100%) tumours were positive for pan-IDH1 expression. EGFR expression was identified in 16/31 (51.6%) tumours and expression was significantly greater in high grade gliomas when compared to low grade tumours (P = 0.04). Furthermore, EGFR expression was significantly greater in gliomatosis cerebri when compared to oligodendroglioma (P = 0.002), astrocytoma (P = 0.01), and oligoastrocytoma (P = 0.04). Ki-67 expression was identified in 28/31 (90.3%) gliomas, and the Ki-67 LI was significantly higher in high grade tumours (P = 0.02). A significant moderate correlation between the Ki-67 LI and EGFR immunoreactivity (r = 0.47, P = 0.007) was identified. The IDH1 R132H point mutation was not identified in this series of canine gliomas, and may not be an appropriate biomarker to aid in the classification and treatment of these tumours. EGFR, however, may be a suitable therapeutic target, particularly for gliomatosis cerebri. The Ki-67 LI may aid in the development of a grading scheme for canine gliomas.
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    Inflammation and endothelial perturbation in canine abdominal surgery: the potential modulatory effect of lidocaine
    Donaldson, Liam Robert ( 2019)
    Complication rates following emergency laparotomy surgery are high, with organ dysfunction being a commonly encountered post-operative complication. Given the endothelium acts as the interface between the systemic circulation and the organs, its function is vital to maintaining organ health. The endothelium is in a constant state of flux, impacted largely by the local environment of which it is a part. In the presence of wide-spread systemic inflammation, inflammatory mediators precipitate change to the structure of the endothelial glycocalyx. These changes result in shedding of the endothelial glycocalyx and alteration of the endothelial phenotype. The endothelium may, as a result, lose the capacity to regulate vasomotor tone, and shift toward a pro-inflammatory and pro-coagulant state. This predisposes to reduced tissue oxygen delivery, and organ dysfunction may ensue. This thesis aimed to answer two key questions: does surgical trauma induced in canine patients undergoing emergent abdominal surgery invoke a systemic inflammatory response and subsequent endothelial activation? And if so, does lidocaine, a proposed immunomodulatory drug, mitigate this effect when given in the post-operative period? Chapter two provides a detailed review of endothelial structure and function, and current literature pertaining to systemic inflammation and endothelial activation in the context of abdominal surgery. Chapter two also examines the literature regarding the proposed mechanisms through which lidocaine acts as an immunomodulatory drug, and reviews publications that investigate the use of lidocaine as an anti-inflammatory drug in human patients after abdominal surgery. Chapter three is a randomized, blinded clinical trial quantifying the effect of emergency abdominal surgery on the concentration of markers of systemic inflammation and endothelial perturbation in canine patients in the post-operative period. The trial also assessed the potential use of lidocaine as a post-operative immunomodulatory therapy in dogs having undergone laparotomy. Fifty canine patients undergoing abdominal surgery were enrolled in the study. Patients were randomized into two separate groups: a study group receiving lidocaine intravenously, and a control group receiving 0.9% NaCl intravenously for a twelve-hour period following abdominal surgery. Blood samples were gathered prior to surgery, followed by six and twelve hours post-operatively. Concentrations of markers of systemic inflammation (IL-6) and markers of endothelial perturbation (VEGF and HA) were quantified via means of ELISA at each time point. Results revealed a significant increase in the concentration of markers of systemic inflammation and endothelial perturbation in post-operative blood samples. No immunomodulatory or endothelial preserving effect of lidocaine was appreciated.
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    Investigations on the pathophysiology of canine idiopathic epilepsy
    Milne, Marjorie ( 2018)
    The causes of canine epilepsy are poorly understood. The current aetiologic classification scheme for canine epilepsy includes the categories structural epilepsy, idiopathic epilepsy with genetic or suspected genetic cause, or idiopathic epilepsy of unknown cause. It is likely that dogs with idiopathic epilepsy of unknown cause have heterogeneous underlying pathologies, including subtle structural change that cannot be identified on conventional visual inspection of brain magnetic resonance images. This thesis examines the causes of epilepsy in a sample of Australian dogs presented to the University of Melbourne Veterinary Hospital. The general hypotheses explored by the studies described in this work are that some cases of canine idiopathic epilepsy have underlying structural brain pathologies similar to those identified in humans with epilepsy, and that canine idiopathic epilepsy is associated with increased cerebral levels of the excitatory neurotransmitter glutamate. The prevalence of epilepsy in dogs presented to the University of Melbourne Veterinary Hospital was 1.1%, and idiopathic epilepsy of unknown cause represented 75% of epilepsy diagnoses. Of the 25% of dogs with structural epilepsy, brain tumours were the most frequent cause (60%), followed by meningoencephalitis of unknown origin (11%). In dogs with idiopathic epilepsy, odds ratio analysis identified 21 breeds of dog with an increased risk of a diagnosis of epilepsy. The Hungarian viszla had a particularly strong association with this diagnosis, but in none of the dogs diagnosed with idiopathic epilepsy was a familial history of epilepsy reported. Based on retrospective, randomised and blinded, subjective review of brain magnetic resonance imaging scans of dogs with idiopathic epilepsy and controls, there was no convincing evidence of the magnetic resonance imaging findings of hippocampal sclerosis or focal cortical dysplasia. Brain MRIs were also evaluated using atlas-based segmentation and volumetry, based on a novel canine brain atlas developed for this research. Hippocampal atrophy was used as a biomarker for possible hippocampal sclerosis. Unilateral or bilateral hippocampal atrophy was identified in 15% of dogs with idiopathic epilepsy, based on identifying those dogs with a hippocampal formation volume below the lower 95% reference limit for hippocampal volume established in control dogs. Increased volume of the cerebral cortex was used as a biomarker for possible cortical dysplasia, and reduced volume of the cerebral cortex was a biomarker for cerebrocortical atrophy. Dogs with idiopathic epilepsy had statistically significant reductions in cerebrocortical volume in the left and right olfactory, temporal, occipital, and right parietal lobes. The second hypothesis was that dogs with idiopathic epilepsy had elevated cerebral glutamate, the major excitatory neurotransmitter in the brain. This was explored non-invasively in dogs with naturally occurring epilepsy, using proton magnetic resonance spectroscopy to measure in vivo brain glutamate. There was no significant difference in either glutamate or glutamate to creatine ratio between dogs with epilepsy and controls. Further work is required to establish whether hippocampal atrophy in dogs with epilepsy is due to hippocampal sclerosis. This work could use both volumetry, T2 relaxometry, and histopathologic evaluation of brain samples. Future investigation of the mechanisms of cerebrocortical atrophy in epilepsy may involve cortical thickness measures to allow targeted correlation of regions of localised cerebrocortical atrophy with brain histology, and fibre tracking to map epileptogenic networks, exploring the connections between EEG identified seizure focus and regions of cortical atrophy. The role of brain glutamate may be further investigated using multi-voxel magnetic resonance spectroscopy at 3 Tesla, and glutamate transporter proteins and receptors may be investigated through laboratory techniques.
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    Viscoelastic coagulation changes in dogs with tiger snake envenomation
    Stanley, Monique Kirstie ( 2018)
    Snake venom induced consumption coagulopathy (SVICC) is an important yet poorly described clinical syndrome in the field of veterinary medicine. Publications referencing humans, on the other hand, are numerous in comparison, especially since the establishment of the Australian Snakebite Project. Since the introduction of Australian snake-specific antivenoms, SVICC has become the most common underlying reason for human fatality from snake envenomation in Australia.1 The annual canine snakebite caseload in Australia alone is vast and could be established as a model for the human condition. The overall objective of this study is to extend the scientific literature on SVICC in veterinary medicine, namely the role of thromboelastography (TEG) in tiger snake envenomed dogs. The study assessed the changing clot kinetics of canine whole blood after natural tiger snake envenomation using an established technique called thromboelastography (TEG). Specifically, we will be determining the TEG changes in dogs at several time points: T0 = time of presentation; T1 = 1 hour after antivenom administration; T18 = 18 hours after antivenom administration; and, finally, T24 = 24 hours after antivenom administration. Tiger snake venom caused alterations in TEG parameters, specifically the prolongation of R time (time to initiation of clot formation) during the first 24 hours of envenomation.2 No clinical benefit exists in using TEG over classical coagulation parameters in the identification of SVICC in tiger snake envenomed dogs. Hypercoagulability occurs for 24 hours after envenomation and VetSVICC appears to resolve 18–24 hours after antivenom, which is suggestive of a shorter period to apparent clinical recovery compared to humans, suggesting that VetSVICC may be a unique clinical syndrome. This project is a stepping stone to future research into both SVICC and tiger snake envenomation in veterinary medicine, including establishing an accessible and reliable diagnostic test for SVICC and further delineation of its importance regarding the severity of illness and outcome.
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    The effects of intravenous fluids on thromboelastrographic variables in dogs
    Chan, Adelina ( 2018)
    Intravenous fluid resuscitation plays a fundamental role in treating dogs in haemorrhagic shock, as it can rapidly replenish lost intravascular volume and improve tissue perfusion. However, a consequence of intravenous fluid therapy is interference with haemostasis, which has a detrimental effect to trauma patients that are already haemostatically compromised. In people, intravenous fluid therapy can effect haemostasis and subsequently can increase haemorrhage, transfusion needs and mortality. The effects of intravenous fluids on haemostasis in dogs have not been widely established. More recently, viscoelastic devices like thromboelastography have been used to examine haemostasis. Compared to conventional coagulation testing, viscoelastic devices have the advantages to be able to assess the speed and kinetics of clot formation, clot strength and even the breakdown of the clot. The aim of our study was to determine the effects of intravenous fluids on coagulation in dogs with the use of thromboelastography. The objective of our study was to determine the effect of dilution of canine whole blood with clinically relevant doses of common intravenous fluids on thrombelastographic variables. Our hypothesis was that in vitro dilution of canine whole blood from healthy dogs with intravenous fluids will induce dose-dependent changes in thromboelastographic variables consistent with hypocoagulability. Further, we hypothesized that the characteristics of the fluids, such as its ionic strength and osmolality, will effect thromboelastographic variables in addition to those of dilution alone. The results of our study showed that in vitro dilution of canine whole blood with commonly used intravenous fluids lead to thromboelastographic changes consistent with hypocoagulability in a dose dependent manner. Besides dilution percentage, viscoelastic changes were influenced by fluid characteristics, specifically ionic strength, osmolality and colloidal properties. In our study, 7% hypertonic saline had the most severe effects on coagulation, followed by 20% Mannitol then 3.4% hypertonic saline. Hydroxyethyl starch 130/0.4 had minimal effects on coagulation besides a dilutional effect. The differential effect of fluid characteristics should be taken into consideration when resuscitating dogs with large fluid volumes, but clinical studies are still required to further delineate the importance of different resuscitation fluids and volumes on haemostasis in dogs.