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Dogs were prospectively recruited following implantation of a St. Jude dual-chamber pacemaker system for 3AVB; dogs with evidence of sinus node dysfunction or clinically important arrhythmias other than 3AVB were excluded. All dogs were programmed to VDD at study entry. The average, lowest and highest heart rates during VDD pacing were determined; the average heart rate was programmed as the base rate for VVI and VVIR. The highest heart rate was programmed as the upper tracking rate for VVIR with a threshold of 2.5, slope of 16, and fast reaction and recovery times. The rest function was used for all pacing modalities and was programmed to the lowest heart rate during VDD pacing; if the lowest heart rate was at the lower rate limit, then the rest rate was programmed 20% slower. Dogs were randomly assigned to VVI or VVIR after initial VDD interrogation and echocardiography (2D, Doppler, TDI). Ambulatory ECGs were recorded for 36 hours and the owners were instructed to record the time and duration of 5 specific activities: (1) urination/defecation, (2) eating, (3) sleeping, (4) walking, and (5) play. After three months, echocardiography was repeated, the dog was crossed over to VVI or VVIR pacing, and ambulatory ECG monitoring was repeated. After three additional months, echocardiography was repeated, the pacing modality was programmed to VDD and ambulatory ECG monitoring was repeated. Atrial rate, ventricular rate, and atrialto-ventricular ratio were determined for each of the 5 activities after an initial 12 hours acclimatization period. This is an ongoing study and currently 6 dogs are enrolled where 5 dogs have completed the VVI phase, 4 dogs have completed the VVIR phase, 4 dogs have completed both the VVI and VVIR phases, and 1 dog has completed all VVI, VVIR, and VDD phases. Atrial rates and atrial-to-ventricular ratios for VVI, VVIR, and VDD at each activity are as follows: urination/defecation (156.0 AE 42.9, 2.01 AE 0.39),(134.4 AE 24.0, 0.95 AE 0.14), (105.7, 1.0); eating (116.4 AE 20.8, 1.51 AE 0.23),(126.2 AE 11.7, 0.97 AE 0.22), (95.7, 1.0); sleeping (99.1 AE 38.3, 1.32 AE 0.62),(79.8 AE 15.7, 0.88 AE 0.30), (51.5, 0.76); walking (183.2 AE 24.7, 2.4 AE 0.41), (157.4 AE 38.9, 1.09 AE 0.12), (91.4, 1.0); play (125.4 AE 38.3, 1.66 AE 0.61),(105.8 AE 17.0, 0.83 AE 0.07), (108.9, 1.0). Echocardiographic data is available for all 6 dogs in VDD, 4 dogs in VVIR, and 2 dogs in VVI. The left ventricle appears to be larger in diastole and systole for VVI (LVAd/Ao = 6.85 AE 0.76, EDV/Ao = 26.08 AE 2.80, LVAs/Ao = 2.74 AE 0.19, ESV/Ao = 9.23 AE 0.08) and VVIR (LVAd/Ao = 5.24 AE 0.57, EDV/Ao = 24.78 AE 3.04, LVAs/ Ao = 2.38 AE 0.47, ESV/Ao = 10.65 AE 3.21) than VDD pacing (LVAd/Ao = 5.14 AE 1.38, EDV/Ao = 23.13 AE 7.46, LVAs/Ao = 2.30 AE 0.84, ESV/Ao = 7.43 AE 4.27).
Medical records of 18 small breed dogs that had bronchial stent placement for chronic coughing secondary to bronchial collapse were reviewed. A hierarchical multiple linear regression analysis was conducted to predict lifespan after bronchial stent placement based on age at time of stent placement, severity of MMVD and the presence of pulmonary hypertension.
Eighteen dogs had bronchial stents placed over a period of 7 years. Age at the time of stent placement ranged from 6.5 years to 14 years of age (M = 10.47 AE 1.85). Breeds represented included Cavalier King Charles spaniel (2), beagle (2), Chihuahua (4), Pomeranian (3), toy poodle (2), Yorkshire terrier (2), Maltese (1), Coton de Tulear (1) and shih tzu (1). There were 11 males and 7 females. Twelve dogs (66.67%) had evidence of moderate to severe MMVD and 4 (22.22%) had evidence of pulmonary hypertension. Six dogs (33.33%) had CHF prior to stent placement and 6 dogs (33.33%) had CHF after stent placement. Syncope was reported in 6 dogs prior to stent placement and 5 dogs after stent placement. The average lifespan after stent placement was 203.56 AE 250.72 days. Three dogs are currently alive post-stent placement (1013, 559 and 411 days).
A hierarchical multiple linear regression was calculated to predict lifespan after placement of a bronchial stent based on age at the time of stent placement, the presence of pulmonary hypertension, and severity of MMVD. In stage one, age at the time of stent placement significantly predicted lifespan after placement of a bronchial stent (b = À7.10, P < .02); with lifespan decreasing by 7.10 days for each additional month of age at the time of stent placement. The presence of pulmonary hypertension did not significantly predict lifespan (b = 235.32, P = .10). Severity of MMVD did not contribute significantly to the model in stage two (b = 101.38, P = 0.43).
Thirty-two dogs of 10 small breeds with varying TRFVs (1.9-5.28 m/s) and PVR (2.9-33.0 WU) were studied. There was a significant effect of observer (P < 0.05), respiration (P < 0.01), and image quality (P < 0.05) on outcome variables whereas imaging position (dogs lying versus standing) did not cause differences (P > 0.20). Sedation significantly increased mean TRFV (P < 0.05, from 3.15 AE 0.71 m/s to 3.51 AE 0.83 m/s; increase observed in 55% dogs) whereas 6-MWT had no effect (3.16 AE 0.75 m/s, P > 0.05). HR declined after sedation (mean reduction 13 AE 19 bpm; P < 0.05) and increased after 6MWT (mean increase 30 AE 28 bpm; P < 0.05) but had no correlation to TVFV or PVR. 6-MWT did not influence TRFV and PVR.
Thirty-two cats were enrolled (25 male, 7 female; all neutered), with a median age of 9.6 years (range 1.1 to 18.0 years). Thirtyone (97%) cats survived to discharge and 28 cats (88%) survived to the 7-10 day re-evaluation. Median NT-proBNP concentrations at admission and hospital discharge were 1,713 pmol/L (range, 160 to 3,784 pmol/L) and 902 pmol/L (range, 147 to 3,223 pmol/L), respectively. Median NT-proBNP concentration at re-evaluation was 1,124 pmol/L (range, 111 to 2,727 pmol/L). Median NT-proBNP concentrations decreased significantly from admission to discharge (P < 0.001) and from admission to re-evaluation (P < 0.001), but not from discharge to re-evaluation (P = 0.28). Median survival time was 109 days (range, 1 to 606 days), with 6 cats (19%) still alive at the time of analysis. Cats that received pimobendan had a significantly greater decrease in NT-proBNP concentrations [À821 pmol/L (range, À2,163 to À267 pmol/L) from admission to discharge compared to cats that did not receive pimobendan [À561 pmol/L (range, À2137 to 889 pmol/L); P = 0.045]. Cats whose owners had difficulty administering medications (n = 11) had a shorter survival time [median 60 days (range, 2 to 234 days)] compared to owners who did not report difficulty [n = 21, median 115 days (range, 1 to 606 days)]; however this was not statistically significant (P = 0.12).
Medical records were reviewed for French and English Bulldogs presented to Oregon State University for heart murmur evaluation or imaging prior to balloon valvuloplasty (BVP). Dogs were included if they had severe PS (pressure gradient >80 mmHg estimated via echocardiography) and confirmation of their coronary artery anatomy by either angiography or computed tomography (CT). Echocardiographic, angiographic, and CT images were reviewed. The pulmonic annulus (PA), aortic annulus (Ao), PA:Ao ratio, valvular anatomy, and coronary anatomy were assessed on each imaging modality by three observers (CS, KS, DDS) and averaged. Tricuspid regurgitation (TR) was assessed on echocardiographic and angiographic studies. Breed medians or means were compared by Mann-Whitney U test or unpaired t-tests as indicated. In the dogs that had both CT and angiographic studies, values were compared by paired t-tests.
Early stages of heart disease are characterized by reduction in energy reserves with impairment of myocardial contractility and relaxation. However, these alterations are difficult to measure with existing clinical imaging modalities such as echocardiography. Recent advances in phosphorous ( 31 P) magnetic resonance spectroscopy (MRS) enable changes in energy reserves to be measured non-invasively.
Utilizing a 7 Tesla (T) MRI scanner, 31 P magnetic resonance spectra were collected with an ungated, free breathing three-dimensional chemical shift imaging sequence. In each dog, three 2.8 cc voxels on the septal wall near midventricle were selected for analysis. The PCr/ATP ratio was computed without saturation or blood correction in each voxel and averaged.
Although there was a range in severity of MR in each group, all dogs were asymptomatic, untreated and stage B1 or B2 during the observation period. All data were collected at each time point. Using each dog as its own control for echocardiographic evaluation, the end-point LVDd increased in diameter (35.1% in 18 weeks dogs, 19.9% in 6 weeks dogs) and left atria dimensions increased (27.3% in 18 weeks dogs and 31.2% in 6 weeks dogs) and the fractional shortening increased (22.6% in 18 weeks dogs, 24.3% in 6 weeks dogs). Compared to baseline, the LVDs dimension increased (18.7% at 18 wks in the 18 weeks dogs, and 8.6% in the 6 weeks dogs). At endpoint catheterization, the forward LV stoke volume decreased (35.2% in 18 weeks dogs, 22.2% in 6 weeks dogs) in all dogs.

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Two groups of dogs of a variety of breeds were recruited including 5 control dogs (euthanized for severe non-cardiac disease; age range 4 months-12 years), and 6 dogs with severe MMVD (euthanized for refractory CHF; age range 9-14 years). Samples were collected from the right atrium (RA), right ventricle (RV), left atrium (LA), left ventricle (LV) and interventricular septum (IVS). Immunohistochemical staining of all tissues with a polyclonal rabbit anti-human antibody against corin was carried out, and computational quantification was used to compare the staining density between the two groups and between cardiac regions.
Our results indicate that corin expression might be a rate-limiting step in dogs with CHF due to MMVD. A lack of an increase in corin expression or activation can be postulated to be a possible important pathophysiological mechanism responsible for the natriuretic peptide paradox in dogs with CHF, and warrants further investigation. Figure 1 : Quantitative corin expression in myocardial tissue in control dogs, and in dogs with congestive heart failure secondary to myxomatous mitral valve disease. Aims of this study were to determine feasibility and repeatability of measuring anatomical regurgitant orifice area (AROA) using real-time 3D transthoracic echocardiography (RT3DE) in dogs affected by MMVD, and to determine if AROA obtained using RT3DE relates to the severity of MMVD assessed both with an echocardiographic scoring system (MRSS) and the American College of Veterinary Medicine (ACVIM) staging system. RT3DE datasets of the mitral valve were acquired in 81 privately owned dogs diagnosed with MMVD and AROA was traced and measured using dedicated software. Dogs were classified as mild, moderate or severe according to an echocardiographic MRSS, and as ACVIM Stage B1, B2 or C. Technique feasibility (as % of dataset measurable) and inter-and intra-operator repeatability were assessed. Differences in AROA between dogs in different MRSS and ACVIM stages were investigated.
AROA was measurable in 60 (74.1%) dogs. The inter-and intra-operator coefficients of variation were 26% and 21%, respectively. AROA was significantly greater in dogs with severe MRSS compared to dogs with mild MRSS (P = 0.04). There were no differences in AROA between dogs with mild versus moderate, and moderate versus severe MRSS (P = 0.84 and P = 0.28, respectively). There was no difference between AROA of dogs in different ACVIM clinical stages (P = 0.17)
In conclusion obtaining AROA using RT3DE is feasible and offers an additional, non-invasive technique to stratify MR severity in dogs with MMVD. The relatively low repeatability suggests standardization of the technique is desirable. Diagnostic and prognostic utility of AROA deserve further investigation. Three-dimensional (3D) saddle shape of the mitral valve annulus was first reported in 1987 by Levine and colleagues. Previous studies showed annulus height to commissural width ratio (AHCWR), an index parameter for saddle-shape of mitral annulus, is preserved across mammalian species (15-20%) . This suggests that nature conserves the saddle-shaped annulus for mechanical benefit. Menciotti et al described 3D transthoracic echocardiography (TTE) of normal dogs and AHCWR was 21 AE 0.05%. There are no reports on 3D transesophageal echocardiography (TEE) studies on dogs with myxomatous mitral valve disease (MMVD). This study aims to evaluate mitral valve annulus morphology using 3D TEE in dogs with MMVD.
Twenty-six dogs were included in the study, 2 dogs were excluded from the study due to the uninterruptable echocardiographic images. Thus, a total of 24 dogs were evaluated in this study. The study group included 7 dogs with ACVIM Stage B2, 10 dogs with Stage C, and 7 dogs with Stage D. Characteristics of each group were summarized below: AHCWR decreased as the disease progressed (Stage B2; 14.2 AE 4.2%, Stage C; 9.0 AE 4.2 5, Stage D; 9.5 AE 3.3%, Figure) . Stage D dogs showed significantly lower values (P < 0.05) compared to Stage B2 dogs. Non-planarity angle measurement tends to increase in value as the severity of MMVD worsened, however, there were no significant differences between the groups (B2; 157.7 AE 8.1°, C; 167.9 AE 9.2°, D; 164.4 AE 8.5°, Figure) . There were no significant differences between the groups for annular circumference and annular area values.

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Six intact hound cross dogs weighing between 19.5-23.8 kg were anesthetized using a standardized protocol and spontaneous ventilation. Each dog underwent MDCT, echocardiography, and TD. Dogs were randomized to the order of MDCT and echocardiography, and TD was performed last. CO using TD was measured in triplicate with <10% variation and minimal changes in heart rate. For imaging methods, CO was calculated by volumetric measurements of SV multiplied by the average heart rate. Echocardiographic left ventricular (LV) end-diastolic volumes (EDV) and end-systolic volumes (ESV) were measured on right long axis 1D images (Teichholz) and right long axis and left apical 2D images (single plane Simpson's method of disks, MOD) on 3 averaged consecutive beats. Real-time 3-dimensional echocardiographic (RT3DE) LV volumes were measured using 3D functional analysis software from both right long axis and left apical imaging planes. MDCT EDV and ESV were measured using specific LV functional analysis software on ECG-gated studies.
These results suggest certain echocardiographic techniques available for CO measurement are comparable to invasively measured CO. Specifically, 2D MOD using the right long axis and left apical 4-chamber views, or RT3DE from the right long axis view, had excellent agreement with TD. The excellent correlation between methods indicates a close relationship between select techniques, although they are not interchangeable. CO is underestimated by the noninvasive techniques studied here compared to TD. The differences between TD and noninvasive measures may be due to errors of the method, patient stroke volume variability, heart rate fluctuation, or inaccuracies of TD. One of the major limitations of this study is the small number of patients, which allows preliminary conclusions only. A larger prospective study is needed to delineate the benefits and constraints of these methods. Myxomatous mitral valve disease (MMVD) is the most common heart disease in the dog. Angiotensin converting enzyme (ACE) inhibitors are frequently recommended for management of dogs with MMVD and asymptomatic cardiac enlargement. However, the benefit of ACE inhibition in dogs before the onset of congestive heart failure (CHF) is controversial, with different studies showing conflicting results. A variable response to ACE inhibitor therapy has also been observed in human beings with heart disease and in some cases this has been attributed to a polymorphism in the ACE gene. We have previously demonstrated a polymorphism in the canine ACE gene, although the clinical significance of this finding is unknown.

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We conclude that dogs that are homozygous for the ACE variant have lower baseline levels of ACE activity but still demonstrate ACE inhibition in response to enalapril. Further study is warranted to evaluate the clinical importance of these findings. The Cavalier King Charles Spaniel is one of the most commonly reported breeds of dogs affected with myxomatous mitral valve disease (MMVD). Angiotensin converting enzyme (ACE) inhibitors are frequently recommended for management of dogs with MMVD. However, the benefit of ACE inhibition in MMVD before congestive heart failure has developed is controversial with different studies showing conflicting results. A variable response to ACE inhibitor therapy has also been observed in human beings with heart disease and has been attributed to a polymorphism in the ACE gene. We have previously demonstrated a polymorphism in the canine ACE gene in small breed dogs.

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PREVALENCE OF DIROFILARIA IMMITIS ANTIGEN IN CLIENT-OWNED PET DOGS BEFORE AND AFTER SERUM HEAT TREATMENT. Laura Nafe, Susan Little, Paul DeMars, Ryan Baumwart, Nalani Yamada, Eileen Johnson. Oklahoma State University, Stillwater, OK, USA Pet dogs are routinely screened for heartworm disease as a component of annual wellness evaluation. Recent evidence suggests that antigen-antibody complexes in canine serum may preclude accurate diagnosis of heartworm disease with commercially available Dirofilaria immitis antigen assays. The objective of this study was to determine whether normal dogs presenting to the Oklahoma State Veterinary Teaching Hospital Community Practice Service for annual veterinary care warrant heat treatment of serum to confirm a negative heartworm antigen test by 4DX Snap (Idexx). Serum and EDTA samples were collected from 201 clientowned pet dogs between December 2014 and October 2015. Dogs ranged in age from 10 months to 15 years (median 4 yrs) and included 108 males and 93 females. Complete survey results (available for 194/201 dogs) revealed 139 dogs (72%) received heartworm preventative regularly over the past year. All EDTA samples were evaluated for microfilaria and serum samples for D. immitis antigen before and after heat treatment using a commericial assay (DiroCHEK, Zoetis). Four dogs (2%) tested positive for D. immitis antigen on 4DX Snap Test (Idexx) and 2 were microfilaremic. On DiroCHEK antigen testing 2 dogs were positive before and after heat treatment, 1 dog was positive after heat treatment only, and 1 dog was negative before and after heat treatment despite being microfilaremic. One dog received heartworm preventative (Heartgard; Merial) regularly (2/12 doses missed). Heat treatment does not appear to improve detection of D. immitis antigen in asymptomatic pet dogs undergoing routine heartworm disease testing with 4DX Snap Test (Idexx). of the mechanisms behind increased concentrations of these cardiac biomarkers in azotemic dogs is warranted for correct interpretation of test results. The aim of this study was to investigate the association between measured glomerular filtration rate (GFR) and plasma concentrations of cTnI and NT-proBNP in stable canine CKD patients.
Median age of included dogs was 6 years (range 0.5-14 years). Median bodyweight was 19.5 kg (range 2-42 kg). There was no difference in age or weight between healthy and CKD dogs. Among dogs with CKD, 16 had a decreased GFR and 11 had a GFR within the normal range. Dogs with CKD and a GFR in the normal range had other evidence of kidney disease such as persistent renal proteinuria or morphological abnormalities.
Univariate analyses were performed to evaluate associations between dog characteristics and cTnI and NT-proBNP, respectively. Age (r = 0.59; P < 0.0001), body weight (BW; r = 0.29; P = 0.04), creatinine (r = 0.29; P = 0.04) and systolic blood pressure (SBP; r = 0.42; P = 0.06) were associated with cTnI concentration. Creatinine (r = 0.46; P = 0.001), GFR (r = À0.39; P = 0.0063), albumin (r = À0.35; P = 0.0141), and erythrocyte volume fraction (EVF; r = À0.35; P = 0.0072) were associated with NT-proBNP concentration.
All variables that were associated with cTnI or NT-proBNP concentration, respectively, with a P-value of <0.2 were included in the multiple regression analysis (except creatinine because of covariance with GFR). Age, BW and SBP were the variables remaining in the final model for cTnI (adjusted R-square 0.50, P < 0.0001). GFR, BW and serum albumin concentration were the variables remaining in the final model for NT-proBNP (adjusted R-square 0.29, P < 0.0001).

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The primary objective was to determine the minimum, average, and maximum heart rate and incidence of arrhythmias for healthy puppies. The secondary objective was to evaluate the correlation between Holter heart rate and age, weight or breed. We hypothesized that healthy puppies will have less arrhythmias and higher Holter minimum, average, and maximum heart rates than healthy adult dogs.
Eligible puppies had to be healthy and between the ages of 12-51 weeks. History, physical exam, and thorough auscultation were performed. An echocardiogram was performed on any puppy with a heart murmur on auscultation. Puppies with physiologic heart murmurs were included in the study while puppies with structural heart disease were excluded. A 24 hours 3-channel Holter monitor was placed on all puppies included in the study. Holter placement followed established methods from the WSU Veterinary Holter Service.
A total of 44 puppies representing 20 breeds were included in this study. Minimum, average, and maximum Holter heart rates were higher in puppies compared to healthy adult dogs. The number of supraventricular complexes seen in puppies were less than in healthy adult dogs. The range of ventricular complexes in puppies were higher than in healthy adult dogs. Minimum and average Holter heart rate decreased with increasing age while maximum Holter heart rate was not correlated with age. No correlation between minimum, average, and maximum Holter heart rates and weight was appreciated. There was a statistically significant difference between maximum Holter heart rate and different breed classifications by size (small versus medium versus large versus giant breed). No correlation between minimum and average Holter heart rate and breed classifications was identified in healthy puppies.
This study is the first study to evaluate normal ambulatory electrocardiographic values in healthy puppies. This information can be used to potentially screen puppies at an earlier age for cardiac disease. The number of ventricular arrhythmias considered normal for healthy adult dogs is thought to vary with dog breed. Whether or not this breed variation is true in puppies is unknown. Now that a normal baseline has been established for puppies, further studies can be conducted to evaluate for breed variation. Patients underwent LUS and TXR within 6 hours. LUS images were scored for presence of B-lines at 4 sites on each hemithorax. An individual LUS site was scored positive if >3 B-lines were observed within a single intercostal space at that site. TXR were scored for presence of alveolar-interstitial infiltrates at 4 sites on each hemithorax, analogous to LUS. An individual TXR site was scored positive if infiltrate was present in least 25% of the site. Medical records were evaluated for final diagnosis. Agreement in distribution of positive sites between LUS and TXR was compared using a Cohen's Kappa coefficient. Patterns of distribution of AIS among different final diagnoses were compared using Fisher's exact tests.

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A minimum of seven dogs in each group (normal versus CHF) were needed to demonstrate the statistical power with a probability of 0.9 and type one error of P < 0.05 based on previous publications. Blood was collected in EDTA tubes from 9 normal geriatric dogs (free of heart murmur and absent history of CHF) and 8 dogs with CHF secondary to MMVD (confirmed by clinical signs, chest x-ray and echocardiography). Plasma samples were subjected to isolation of miRNAs with a miRNeasy Plasma Kit (Qiagen) and subsequently reverse-transcription reaction with a miScript II RT Kit (Qiagen). Validated canine primers of 12 candidate miRNAs and miR-39 (spike-in control) were employed to quantify relative expression levels of each miRNA between normal and CHF dogs via real time PCR (miScript SYBR Green PCR Kit, Qiagen).
Canine specific miRNAs were successfully isolated from plasma and amplified. Plasma miR-21 (P = 0.015), miR-133 (P = 0.01), and miR-1 (P = 0.01) were differentially expressed in CHF dogs with statistical significance when compared to normal dogs. These miRNAs are known to be involved in extracellular matrix remodeling and the regulation of fibrotic genes such as TGF-b and Smad. The results suggest that alterations in the interaction between miRNAs expression profiles and their downstream target genes may play a role in cardiac progression to CHF in dogs with MMVD. Further investigation remains warranted to determine their diagnostic potential as molecular biomarkers for early detection of CHF in dogs with MMVD. In order to assess signalment and concurrent disease processes in dogs diagnosed with aortic thrombotic disease, a retrospective review was performed of records available through the Veterinary Medical Database of dogs examined between 1985 and 2011. Information on age, sex, breed, weight, and concurrent disease processes in dogs reported with AT were reviewed. Five control dogs without a diagnosis of AT were selected for each AT case.

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AT was diagnosed in 291 of 984,973 dogs included in the VMDB from 1985 to 2011. The odds of a dog having AT did not differ significantly by sex (P = 0.0593), age (P = 0.9833), or weight (P = 0.7766). Compared to mixed breed dogs, Shetland sheepdogs had a significantly higher odds (OR=2.44, P = 0.0027) of being diagnosed with AT. Protein-losing nephropathy was the most commonly reported disease occurring concurrently in dogs with AT (64/291).
Signalment data including age, sex, and weight were not significantly different in dogs diagnosed with AT compared to dogs not diagnosed with AT. Dogs with AT were commonly diagnosed concurrently with PLN. Contrary to previous reports, cardiac disease was not a common concurrent diagnosis in the dogs with AT included in this study. The function of right ventricle (RV) has gained increased recognition over the past years. In pulmonary hypertension (PH) patients, this led to the reconceptualization of the RV as part of right ventricular pulmonic circulation unit and the acknowledgement that RV function is a major determinant of prognosis in pulmonary arterial hypertension. With this in mind, we sought to investigate the RV systolic function in dogs with post-capillary PH ascribed to mitral valve disease (MVD). Twenty nine dogs with PH (2.5-17.2 kg; 7-15 years) and 51 dogs without PH (1.5-32.0 kg; 2-16 years) were recruited into this prospective cross-sectional observational study. The PH group was subdivided into mild PH (n = 19), moderate PA (n = 6), and severe PH (n = 4). Several breeds were represented. All animals underwent a complete echocardiogram, which included the measurement of RV fractional area change (RV FAC); tricuspid annular plane systolic excursion (TAPSE); RV free-wall longitudinal systolic velocity (tricuspid annular S´wave), and RV fractional shortening (RV FS % ). For improved statistical analysis and normalization of data according to animal size, a body weight-indexed TAPSE (TAPSE BW-indexed ) was created. Some standard echocardiographic indices of congestion and LV function, including early-to-late LV filling velocities ratio (E/A), isovolumic relaxation time (IVRT), E-to-IVRT ratio (E/IVRT), left atrium-to-aorta ratio (LA/Ao), and LV fractional shortening (LV FS % ) were recorded as well. The data was not normally distributed. The Mann-Whitney test showed no differences (P > 0.05) to exist for RV FAC, RV FS % , TAPSE, TAPSE BWindex , and tricuspid annular S´wave between dogs with PH and those without PH. Also, the analysis of the subdivided PH group showed no statistical difference between the three degrees of PH. Spearman correlation coefficients between RV systolic data and LV indices of congestion and function showed a significant positive correlation between TAPSE and E/IVRT (R = 0.3917, P = 0.0003), TAPSE and LA/Ao (R = 0.3298, P = 0.0029), and TAPSE BW-indexed and LV FS% (R = 0.2757, P = 0.0166). Finally, a significant positive correlation was documented between RV FAC and RV FS % (R = 0.3507, P = 0.0100), as well as between TAPSE and tricuspid annular S´wave (R = 0.4575, P < 0.0001). Because only a few animals with severe PH were recruited into this study, the absence of association between the severity of PH and the identification of RV systolic impairment needs further investigation. Nevertheless, we observed a downward trend for RV FS %, TAPSE, TAPSE BW-indexed , and tricuspid annular S´wave along with the aggravation of PH. Our study found correlations between a few congestion and LV function parameters, which may prove useful in monitoring RV function in dogs with severe MVD and overt signs of post-capillary PH. Since the majority of PH cases were mild, it is likely that only a few had reactive PH, which may explain why RV function of either group behaved very similarly. Also, it is not clear whether RV remodeling and impairment would be stronger if pre-capillary PH dogs had been included in this investigation. Decreased heart rate variability (HRV) has been found in dogs in congestive heart failure (CHF) and previous studies have acknowledged the use of HRV indices as prognostic indicators in patients with mitral valve disease (MVD). The vasovagal tonus index (VVTI) is an unconventional time domain indicator of HRV, which is mainly influenced by cardiac parasympathetic tone. In this cross-sectional observational study, we sought to investigate the VVTI in dogs with MVD. Electrocardiographic recordings of 30 dogs (7-16 years, 3.5-15.5 kg) previously classified into ACVIM stages A (controls), B, or C (10 dogs each group) were used to calculate the VVTI. For this, 20 consecutive R-R intervals were measured from each ECG recording (R-R1 to R-R20), and the index was obtained from the formula VVTI = LN [VAR (R-R1 -R-R20)], where LN: natural logarithm and VAR: variance. Bad quality ECG tracings and recordings from dogs with non-sinus rhythms or animals undergoing anti-arrhythmic treatment were not included in this investigation. Also, we recorded the BW-indexed LV in diastole and systole, wall stress index in diastole and systole, fractional shortening, left atrium-to-aorta ratio, mitral E wave, mitral E/A, isovolumetric relaxation time, and the E-to-IVRT ratio. All data underwent the Shapiro-Wilk test to check for normal distribution, while ANOVA, followed by Tukey's test, were used to compare the VVTI between groups. Pearson's test was used to search for linear correlations between the VVTI and the echocardiographic data. The mean values (with lower and upper 95% CI of mean) of VVTI for dogs in stages A, B, and C were, respectively: 8.45 (7.36 -9.54), 6.09 (4.47 -7.71) and 6.34 (5.01 -7.68). A significant difference was found between groups (P = 0.0189), with the mean VVTI being significantly higher in control animals as compared to dogs with stage B MVD (P < 0.05). When it comes to the relationship between VVTI and cardiac rhythms, a significant difference existed between animals in sinus rhythm (SR), sinus arrhythmia (SA), and sinus tachycardia (ST) (P = 0.0083). The lowest VVTI was documented for dogs in ST (5.82; 95% CI 4.14 -7.49), while the higher was found for animals presenting SR (8.27; 95% CI 7.40 -9.15). Significant negative correlations were found between VVTI and LA/Ao (R = À0.3699; P = 0.0443), as well as between VVTI and heart rate (R = À0.4864; P = 0.0064). Although no correlation existed between body weight and the VVTI, age and VVTI attained a significant negative correlation (R = -0.3827; P = 0.0369). The negative correlation between VVTI and heart rate is likely ascribed to the role played by the parasympathetic tone in VVTI, therefore producing higher values when slower rates and irregular rhythms are present. Even though uncontrolled conditions during ECG recording, including stress and agitation, may increase HR, the lower VVTI found in animals exhibiting sinus tachycardia may suggest a sustained sympathetic activation. Although further investigation is warranted, the confidence intervals of this study point to the VVTI <5 being a potential prognostic indicator for CHF, whereas a VVTI >7.7 is likely suggestive of reduced risk for congestion. This is supported by the correlation between VVTI and LA/Ao, as well as the difference between the means of control dogs (stage A) and stage B MVD animals. (6%), Poodle (6%) and Dachshund (6%), besides a mongrel dog (6%). There was a predominance of female dogs (71%). The most common clinical manifestations observed were syncope (71%), presyncope (24%), dyspnea (18%) and exercise intolerance (12%). Twelve percent of the dogs were asymptomatic. The most frequent electrocardiographic changes found were sinus arrest (75%), junctional escape rhythm (73%), supraventricular tachycardia (53%) and sinus arrhytmia (53%). Forty-seven percent of the dogs presented tachycardia-bradycardia syndrome. Only 24% of the animals received pacemaker implants, given the high percentage of animals unable to go through chirurgical/anesthetic procedures (29%), and the presence of asymptomatic animals. The dogs that received pacemaker implants lived for 20.4 months (mean value), whereas the ones that were treated conservatively lived for 10.8 months (mean value). These values showed benefit to those patients who received pacemaker implants, but was hindered by the high percentage of comorbidities in older animals, besides the small number of patients enrolled in this retrospective study.

C27 THE UTILITY OF SPECTRAL DOPPLER OF THE HEPATIC VEINS IN DOGS WITH TRICUSPID REGURGITATION.

Reveal LINQTM wireless cardiac device were obtained via donation following explantation from human patients and the battery life assessed prior to use in each canine patient using a Medtronic CarelinkÒ programmer with appropriate software loaded. An area on the left lateral thorax was clipped and alcohol was used to remove oils. Adhesive electro-conductive gel pads were applied underneath the device and it was placed within the 4th intercostal space parallel to the ribs and then slowly moved in a grid pattern to find the most consistent electrocardiogram (ECG) on the programmer screen. The device was then secured to the thorax using TegadermTM film, gauze, and ElastikonÒ. It was then taped around the thorax with 2 inch white tape and VetrapTM and the patients were fitted with a Holter monitor vest to reduce likelihood of device tampering or consumption of bandage materials and device. The device was programmed with the patient's information and the pre-programmed manufacturer heart rate parameters Patients wore the ICM for 4 to 5 days and owners were asked to activate the monitor manually for 4 total recordings during different activities and to keep a diary. Accuracy was assessed based on quality of ECG tracings (consistent ability to measure heart rate, visualization of entire p-QRS-T waves) and presence of a recording during manual activation. Two dogs were fitted with 24 hour Holter monitors as a control.
This study investigated the relationship between the measured AoSA and response to combined cutting and high pressure balloon valvuloplasty (CB/HPBV), in dogs with severe SAS. Retrospective evaluation of angiographic and echocardiographic video loops of 22 client-owned dogs of various breeds affected with severe SAS (PG mean 143.36 mmHg; range 80.49-322.5 mmHg) who underwent CB/HPBV were evaluated at the baseline time point immediately prior to the procedure. Two board-certified veterinary cardiologists and 1 novice evaluated these video loops to identify still frame images to be used to measure the AoSA. Each evaluator was blinded to the other 2 evaluators' selected images and subsequent measurements of the AoSA. All evaluators were blinded to initial PG and patient outcomes following the procedure. The percentage decrease in PG at 24 hours, 6 months, and 12 months after ballooning was then calculated and correlated to the measured AoSA at each recheck and for each imaging plane.
Medical records and echocardiographic studies from 2008-2015 were reviewed and measured. Cats were categorized into four groups: 1) L-CHF cats with pleural effusion, with or without pulmonary edema, 2) L-CHF cats with pulmonary edema only, 3) cats with pulmonary disease without evidence of cardiomyopathy, and 4) healthy cats. Pulsed wave Doppler recordings of pulmonic outflow were used to measure acceleration time (AT), ejection time (ET), AT:ET ratio, and maximum velocity through the pulmonic valve (PA Vmax). The main pulmonary artery diameter (MPA), aortic diameter (Ao), MPA:Ao ratio, left atrial diameter (LA), and LA:Ao ratio were measured from right parasternal short axis images, while right atrial (RA) maximum diameter, left atrial maximal diameter (LA Long ), RA:LA Long ratio, and RA:Ao ratio were measured from right parasternal long axis images. Studies were reviewed for the presence of tricuspid regurgitation (TR) and the maximum TR velocity (TR Vmax) was measured when available. Groups were compared by one-way ANOVA or Kruskal Wallis tests where appropriate.

N01

Nine client-owned DPs that had a neurologic examination consistent with a cervical myelopathy were prospectively evaluated. All dogs underwent standard MRI in a neutral position using a 3.0 T magnet. Following standard MRI, the patients were placed in right lateral recumbency on a positioning device and additional MRI sequences were obtained with the cervical vertebral column positioned in flexion and extension. Morphologic and morphometric assessments were performed using the neutral, traction, flexion and extension images. Morphologic analysis included a modified spinal cord compression score (previously published), direction of spinal cord compression, signal intensity changes within the spinal cord, and worst site of spinal cord compression. Morphometric assessment included spinal cord height, intervertebral disc width, spinal cord width, spinal cord area, and vertebral canal height. A Fischer's exact test was used to evaluate the morphologic data and mixed-effects linear regression was used to analyse the morphometric data. Interobserver and intraobserver analyses were performed.
Due to space constraints, only part of the results is presented. With neutral positioning, 14 total compressions were noted; 5/9 patients had 1 compression, 3 patients had 2 compressions, and 1 patient had 3 compressions. With flexion, 12 total compressions were noted; 1 patient had no compressions, 5 patients had 1 compression, 2 patients had 2 compressions, and 1 patient had 3 compressions. With extension, 26 total compressions were noted; 1 patient had 1 compression, 6 patients had 2 compressions, 3 patients had 3 compressions, and 1 patient had 4 compressions. In neutral, the majority of lesions were mild (11/14) and located ventrally (9/14). Extension was associated with worsening of compressions in 7 out of 14 sites and the presence of severe compressions (4/26). It was also associated with multiple directions of compression; there was ventral and dorsal compression at 12/26 sites with extension. Morphometric results revealed a decrease in spinal cord height at C6-C7 (midsagittal images) from neutral to extension (P = 0.003) and from flexion to extension (P = 0.001).
This study supports the use of kMRI to evaluate the dynamic component of DA-CSM. Results suggest that extension is most useful; not only did it demonstrate worsening of multiple compressions but it was also able to detect new compressions not visualized in a neutral position. SPACE (Sampling Perfection with Application optimized Contrasts using different flip angle Evolutions) is a T2-weighted threedimensional magnetic resonance imaging sequence. Currently, 2 dimensional T2-weighted spin echo imaging is the gold standard diagnostic imaging for thoracolumbar intervertebral disk disease. SPACE is potentially advantageous to standard MRI in that it is a single acquisition, faster, technically easier modality with higher resolution and the potential to reconstruct the image set in any plane of section. The purpose of this study is to determine the agreement between SPACE and T2-weighted standard spin echo imaging sequences in the presence of Hansen Type I intervertebral disk disease in the thoracolumbar spine of dogs to determine if SPACE is a viable imaging sequence in the canine patient. Our hypothesis was that the MRI sequence SPACE is able to positively identify Hansen Type 1 intervertebral disk disease with equal accuracy to standard T2-weighted spin echo imaging.

N02

This was a double blinded controlled study of dogs with an acute onset T3-L3 compressive myelopathy secondary to Hansen Type I intervertebral disk disease. All patients had standard preanesthetic screening and underwent magnetic resonance imaging of the T3-L3 spine using a Siemens 1.5T scanner. Images were obtained, in at least, the following sequences: standard T2-W sagittal and transverse, STIR dorsal, and SPACE. All dogs underwent a decompressive hemilaminectomy based on the imaging findings. The MRI sequences were randomized and three blinded individuals interpreted the MR images a total of three times. The evaluators were of different skill levels, including one ECVN diplomate, one ACVR diplomate, and a third year neurology resident. The standard T2-weighted and SPACE sequences were individually evaluated for site, side, and degree of compression as well as surgical recommendation and subjective confidence of imaging findings. 23 dogs met the inclusion criteria. There was statistically significant agreement for both site and side (P-Value <0.001) between the standard T2-weighted spin echo imaging and SPACE. There was complete agreement by all three researchers concerning both the site and side of disk herniation (comparing standard T2weighted standard spin echo imaging and SPACE) for 20 of the 23 dogs, and partial agreement for the other 3 dogs. Compressive Length Ratio (CLR) estimates by the two methods were similar, with more variability due to evaluator variability than to differences between the two systems, and similar internal (intra-rater) variabilities between the two systems. The degree of compression, as measured by the standard mild/medium/severe rating protocol shows significantly positive association between the two methods, although not as strong as that between the two methods for the CLR measurements.

N04

Our in vitro data suggest that MBZ and FBZ may be good candidates for treatment of canine glioblastomas. Further in vivo studies are required. Cerebrospinal fluid (CSF) flow in the subarachnoid space can be impeded by developmental anomalies of the subarachnoid space (i.e. diverticula) or acquired adhesions in older animals. Developmental lesions can occur in any type of dog and clinical signs typically present in animals less than one year old; acquired lesions appear to be most common in middle-aged and older small dogs, notably pugs. Clinical signs include progressive proprioceptive ataxia and, at later stages, fecal and urinary incontinence. Traditional surgical treatment of acquired subarachnoid adhesions (constrictive myelopathy) is associated with unreliable outcomes. Here we report on the surgical procedure and outcome using subarachnoid-subarachnoid shunting as a means of bridging the area of adhesion.
The purpose of this pilot study was to determine levels of 11dehydro-thromboxane B 2 (11TxB 2 ) (a stable hydrolysis product of TxA 2 ) in dogs with a variety of neurological and non-neurological diseases, using an enzyme immunoassay. Urine, serum, and CSF samples were stored immediately following collection at À20°C (or at 4°C for up to 7 days prior to À20°C storage).
11TxB 2 was reliably detected in urine, but not in serum or CSF. Storage at 4°C up to 7 days did not significantly affect urine TxB 2 levels, and concurrent corticosteroid or non-steroidal anti-inflammatory medication did not apparently affect levels within the study group as a whole. Overlap of 11TxB 2 urine levels was seen between clinically normal and neurological animal groups. Although some of the highest 11TxB 2 values were seen in animals with intracranial neoplasia, overlap with other neurological disease groups was present. Assay of 11TxB 2 is feasible; however, additional analysis of a large study population will be necessary to determine whether a threshold urine 11TxB 2 level may be useful as a diagnostic biomarker. Canine degenerative myelopathy (DM) is an adult-onset neurodegenerative disorder. Clinical progression is homogeneous within and across breeds, thus four distinct disease stages have been identified (Kanazono et al., 2013; Coates and Wininger, 2010) . DM shares similarities with some forms of amyotrophic lateral sclerosis (ALS), including underlying mutations in superoxide dismutase-1 (SOD1). SOD1-mutant microglia play a central role in ALS progression in rodent models. Microglia behavior is complex, as expression patterns of classic M1 and M2 molecules are variable throughout disease and between species studied. The mechanism(s) underlying microglial phenotype determination within ALS is not known. Fractalkine, a neuronally produced chemokine, has been shown to suppress SOD1 mutant microglial-mediated neurotoxicity in vitro. Thus, fractalkine is a possible contributor to microglial phenotype determination in ALS. Increased spinal cord microglia have been documented in end-stage DM-affected dogs, however, their response and phenotype throughout disease progression is unknown. The goals of this study were to (1) quantify and phenotype microglial cells in close proximity to lumbar spinal cord motor neurons of DM-affected dogs at each disease stage and compare these findings to age-matched, neurologically normal dogs and (2) correlate microglial phenotype with fractalkine protein levels in lumbar spinal cord throughout disease progression.
Spinal cord tissue was collected post-mortem from client-owned dogs donated to the University of Missouri for research purposes. Lumbar spinal cord microglia analyses were conducted with immunofluorescence and confocal microscopy. To identify microglia in close proximity to motor neurons (MNs), concentric circles with increasing radii of 6 lM were applied to MNs of DMaffected dogs at each disease stage (stage 1 n = 120; stage 2 n = 118; late DM (3&4) n = 149) and age-matched control dogs (n = 145). Cells with positive immunoreactivity for Iba-1/iNOS (M1 microglia), or Iba-1/arginase-1 (M2 microglia), were quantified in a double-blind manner. Data were analyzed via Kruskal-Wallis ANOVA on ranks with post-hoc Dunn's method. Fractalkine total protein was quantified via western blot analysis of lumbar spinal cord from DM-affected dogs (stage 1 n = 2; stage 2 n = 4; late DM n = 3) and age-matched control dogs (n = 3). Relative optical densities were normalized to control dogs. Data were analyzed via ANOVA with post-hoc Holm-Sidak method.
A progressive increase in total microglia closely associated with lumbar motor neurons was observed in DM-affected dogs (stage1: 19 AE 0.7; stage 2: 25 AE 0.9; late DM: 28 AE 0.9; P < 0.05) compared to age-matched controls (13 AE 0.4; P < 0.05). Total M2 microglia were increased in stage 2 (7.5 AE 0.5) and late DMaffected dogs (6.2 AE 0.5) compared to age-matched controls (3.3 AE 0.7; P < 0.05). No differences were observed in total M1 microglia throughout disease progression. Lumbar spinal cord fractalkine protein was increased in stage 1 (2.2 AE 0.2) and late DM-affected dogs (2.2 AE 0.3) compared to age-matched controls (1 AE 0.1; P < 0.05).
These data suggest that M2, but not M1, microglia are progressively recruited to motor neurons during disease progression. Increased lumbar spinal cord fractalkine correlates with progressive M2 microglia in DM-affected dogs. Canine degenerative myelopathy (DM) is an adult-onset neurodegenerative disorder with four defined disease stages. DM initially manifests as spastic upper motor neuron paraparesis and general proprioceptive ataxia (stage 1). Progressive neurodegeneration results in non-ambulatory paraparesis/paraplegia (stage 2) and thoracic limb paresis (stage 3). End-stage disease culminates in flaccid tetraplegia, widespread muscle atrophy and signs of brainstem dysfunction (stage 4). The clinical spectrum of DM is homogeneous within and across breeds. Phosphorylated neurofilament heavy (pNF-H), a major structural protein of myelinated motor axons, has shown promise as a prognostic biomarker in other diseases of the nervous system such as amyotrophic lateral sclerosis (ALS). The goals of this study were to 1) quantify the concentration of pNF-H in the CSF of DM-affected dogs throughout disease progression and compare these finding to age-matched, neurologically normal dogs with and without SOD1 mutations and 2) compare CSF concentrations of pNF-H between DM-affected and dogs with other chronic spinal cord disease that mimic DM.
Using an enzyme-linked immunosorbent assay (ELISA), we measured pNF-H concentration in CSF from aged control dogs (n = 10) and DM-affected dogs at all stages (stage 1 n = 9; stage 2 n = 6; stage 3 n = 6; stage 4 n = 3). Data were analyzed using Kruskal-Wallis ANOVA on ranks with post-hoc Dunn's method. Additionally, CSF pNF-H concentration was compared between DM-affected dogs (n = 24) and dogs with other chronic spinal cord disease (n = 7). Those data were analyzed by the student's t test. To evaluate diagnostic performance, a receiver operating characteristic curve was generated.
CSF concentrations of pNF-H were increased in DM-affected dogs at all disease stages compared to aged control dogs (P = 0.002); pNF-H concentrations did not differ between DM disease stages (P = 0.704). DM-affected dogs had increased CSF pNF-H compared to dogs with other chronic spinal disease (P < 0.001). Using a cut-off concentration of 1.5 ng/mL, sensitivity and specificity for diagnosis of DM were 0.83 and 0.95, respectively. Area under the receiver operator characteristic curve was 0.911.
These preliminary findings suggest that pNF-H in CSF is a sensitive and specific disease marker for DM. These data warrant further study of pNF-H in CSF and serum for diagnosis of disease and longitudinal monitoring of therapeutic efficacy in DM. Juvenile onset seizures are defined as seizures starting prior to 12 months of age for this study. The aims of this study were to 1) identify the common etiology and 2) describe the outcome (dead or alive) in cats with juvenile onset seizures. Inclusion criteria were 1) a confirmed or suspected seizure before 12 months of age and 2) complete medical records including a final diagnosis. The diagnosis was reclassified for each cat according to the 2010 ILAE guidelines. Fifteen cats met the inclusion criteria. Median age at onset was 24 weeks (range, 0.4 -40 weeks). Six cats (40%) were diagnosed with structural epilepsy, 5 cats (33%) were diagnosed with unknown epilepsy, and 4 cats (26%) were diagnosed with reactive seizures. Generalized seizures were documented in 9 cats, 4 cats had focal seizures and 2 cats had both focal and generalized seizures. Six cats (40%) had cluster seizures, and status epilepticus was documented for 1 cat. Overall 8 cats (53%) were alive, 6 cats were dead and 1 cat was lost to follow up. (Follow up 16 -173 months after diagnosis) Three cats died, or were euthanized, secondary to seizures or the underlying diagnosis and 3 cats died or were euthanized unrelated to their seizures or seizure diagnosis and 1 cat died due to unknown causes. Primary genetic epilepsy is the most common cause of seizures in dogs, reportedly affecting 0.5-5.7% of the population. There have been several investigations regarding the serum concentrations of trace nutrients, including copper, selenium, zinc, manganese, and iron in human epileptics and animal models. However, no research of this nature is available in dogs with primary genetic epilepsy. This is a prospective pilot study, designed to compare the serum concentrations of several trace nutrients in non-epileptic dogs compared to dogs with epilepsy.
We obtained fecal samples from dogs diagnosed with MUO and from controls, many of which were matched for breed, age and gender. Dogs that had been treated with antibiotics within the previous 4 weeks were excluded. The fecal microbiome was characterized using 16S rRNA sequencing by Illumina and qPCR assays targeting Faecalibacterium spp, and abundances were compared between affected dogs and controls.

N14

All but one dog became or remained ambulatory without assistance. One dog diagnosed with presumptive FCEM presented paraplegic without nociception, did not recover an ambulatory status after 310 days and was euthanized. Only 20% dogs with ANNPE and 14% dogs with presumptive FCEM recovered a completely normal gait according to their owners, including 29% of the grade 2/5 dogs, 12.5% of the grade 3/5 dogs and none of the dogs that were plegic at presentation. The percentage of dogs with persistent motor deficits was not statistically different between the two groups. The time to recovery of some motor function in the most affected limb when presented with mono-or paraplegia was significantly longer in dogs with presumptive FCEM compared to dogs with ANNPE (P = 0.025); however only two cases with this variable were present in the FCEM group. Dogs diagnosed with presumptive FCEM were significantly younger (P = 0.025) and lighter (P = 0.037) than dogs with ANNPE and had a longer intramedullary hyperintense lesion in sagittal T2weighted MRI images (P = 0.001). Dogs with ANNPE were more frequently reported to be performing physical activity (P = 0.001) and to vocalise (P < 0.0001) at the time of onset of the clinical signs and more frequently received anti-inflammatory drugs after diagnosis (P = 0.012) compared to dogs with presumptive FCEM.
In conclusion, dogs diagnosed with ANNPE affecting the thoracolumbar spinal cord segment developed impaired urinary/fecal continence significantly more frequently than dogs with presumptive FCEM. Impaired urinary/fecal continence can occur also in dogs that are ambulatory at presentation. The presence of persistent motor deficits is extremely common, particularly in dogs that presented non-ambulatory; however, no statistically significant difference was detected in the degree of recovery of motor function between the two groups. Morphological abnormalities of the caudal fossa are increasingly recognized as a cause of morbidity in many brachycephalic dogs but there is little information about these conditions in cats. The objective of this study was to investigate presence of similar morphological abnormalities of the caudal fossa of mesaticephalic and brachycephalic cats and possible association with clinical signs and final diagnosis. Feline brain MRI studies at two referral centres were reviewed for cases without visible disease that could alter anatomic landmarks or raise intracranial pressure. T2 weighted median plane images of the brain and if available spinal cord were reviewed for the presence of caudal cerebellar indentation, coning, herniation, syringohydromyelia, and fluid accumulation in the middle ear. Area of the cerebral hemispheres and cerebellum was also measured Blind measurements were taken by two observers and reviewed.
There were 53 feline MRI studies meeting the inclusion criteria. Cerebellar herniation was found in 48% of mesaticephalic and 78% of brachycephalic cats. Syringohydromyelia was not identified in any of the MR studies including only the first two cervical spinal cord segments (36 cats) or more regions of the spinal cord (17 cats). High signal on T2-weighted transverse images of the middle ear was detected in 8% of mesaticephalic and 17% of brachycephalic cat. Information on presenting complaint, results of neurological examination and final diagnosis in cats with and without cerebellar herniation was reviewed.
Mesaticephalic breed of cats show indentation of the caudal aspect of the cerebellum and caudal cerebellar coning similar to brachycephalic breeds of cats. The dimensions of the foramen magnum planum and the area of the cerebral hemispheres and cerebellum measured on a T2W sagittal MR image of the brain were also similar between the two groups. However, brachycephalic cats had a higher percentage of herniation of the cerebellar vermis through the foramen magnum. Herniation of the cerebellar vermis does not appear to be associated with syringomyelia in cats. Feline hyperesthesia syndrome (FHS) was first reported in 1979 and described as an accentuation of an otherwise normal behavior in cats consisting in episodes of tail chasing, biting or licking the lumbar area, flank, anal area or tail; skin rippling and muscle spasms; excessive and unusual vocalizations, wild and uncontrolled jumping and running and presumed hallucinations. FHS has been anecdotally reported in association with dermatological, behavioural, orthopaedic and neurological conditions. Treatment with phenobarbital, gabapentin, prednisolone, amitriptyline, fluoxetine, clomipramine and multivitamin supplements have been used with inconsistent responses. However, we are not aware of any clinical scientific investigation on Feline Hyperesthesia Syndrome.

N16

Thirteen cats were included in the study. The median age of presentation was 1 year (1-7 years). Eleven cats were attacking or over-grooming their tails and two cats their flank or perineal area. Tail mutilation was reported in 9/13 cats, rippling of the thoracolumbar skin in 10/13 cats and 6 cats presented with unusual vocalization during the episodes. The episodes occurred multiple times per day in 12/13 of the cases, and multiple times per week in 1/13. No consistent triggers were reported. Results of CBC and biochemistry were available for 12/13 cats, spinal radiographs for 7/13, magnetic resonance imaging for 6/13, cerebrospinal fluid analysis for 3/13, electromyography for 3/13, dermatological assessment for 7/13, and joint fluid analysis for one cat. The diagnostic work up led to the diagnosis of allergic dermatitis in 2 cases and immune-mediated polyarthritis in one case, but a definitive diagnosis was not reached in the remaining 10 cases. Treatment included corticosteroids in 7/13 of the cats, gabapentin in 8/13, clomipramine in 4/13, topiramate in 2/13, phenobarbital in 2/13 and cyclosporine in 6/13. The majority of the cases (10/13) received a combination of 2 or more drugs.
Our study confirmed previous anecdotal reports of FHS affecting young cats and the possibility of that FHS signs are secondary to inflammatory skin conditions. A definitive diagnosis was not reached in 10 cases. However, not all the cases had the same diagnostic work up and not all had a full behavioral and dermatological assessment. Based on the results of this retrospective study, specialists in neurology, behavior and dermatology have formulated a standardized questionnaire and proposed a rational diagnostic approach for investigation and treatment of this constellation of clinical signs in cats and as a platform for future prospective studies. Although pelvic limb paralysis can be partially circumvented by using carts to allow greater mobility, care of the urinary bladder is often more problematic. In addition to urinary incontinence and recurrent urinary tract infections, it is possible that many such affected animals can suffer from detrusor hyper-reflexia. This is a risk factor for chronic elevation of bladder pressure and secondary renal damage in both human and canine spinal cord injury patients.

N17

Several cystometric metrics were measured in this study: (1) intravesicular pressure at leak; (2) bladder holding capacity expressed as a percentage of the physiologic maximum holding capacity of 20 mL/kg; and (3) bladder compliance, which was determined as change in bladder volume divided by change in intra-vesicular pressure. Two standard points were used for the calculation of bladder compliance: (1) the intra-vesicular pressure and bladder volume at the start of bladder filling (both were zero); and (2) the intravesicular pressure and bladder volume at cystometric capacity or immediately before the start of any detrusor contraction that resulted in urine leakage.
We conclude that there is evidence that detrusor hyper-reflexia occurs in a small proportion of pet dogs paralyzed because of severe T3-L3 spinal cord injury. It is characterized by elevation in intravesicular pressure and reduction in bladder compliance and bladder holding capacity. This identification of a specific type of neurogenic bladder dysfunction permits timely interventions to mitigate renal damage. Potential therapies for treating detrusor hyper-reflexia can now be evaluated using these quantifiable bladder functional parameters derived from cystometry. Osseous-associated cervical spondylomyelopathy (OA-CSM) is a condition characterized by static and dynamic spinal cord compressions. The dynamic component of cervical spondylotic myelopathy in humans (hCSM) is evaluated with kinematic MRI (kMRI). The purpose of this study was to evaluate kMRI in dogs with OA-CSM using a positioning device that allowed controlled flexion and extension of the cervical vertebral column. Our hypothesis was that kMRI would reveal new compressive lesions not identified with standard positioning.
Ten Great Danes and 2 Doberman Pinschers with neurologic examination findings consistent with a cervical myelopathy were prospectively evaluated. All dogs underwent standard MRI in a neutral position (sagittal and transverse T1 and T2 weighted images) using a 3.0 T magnet. The patients were then placed in right lateral recumbency on a positioning device and the cervical vertebral column was first flexed and then extended. Sagittal and transverse T2 weighted images were acquired in both flexion and extension. Morphologic and morphometric assessments of the cervical vertebral column in neutral, flexion and extension were performed. Morphologic analysis included a modified spinal cord compression score (previously published), direction of spinal cord compression, signal intensity changes within the spinal cord, and worst site of spinal cord compression. Morphometric assessment included spinal cord height, intervertebral disc width, spinal cord width, spinal cord area, and vertebral canal height. A Fischer's exact test was used to evaluate the morphologic data and mixedeffects linear regression was used to analyze the morphometric data. Interobserver and intraobserver analyses were performed.
Our results support the use of kMRI in patients with OA-CSM to reveal new compressive sites, dorsal compressions and to enhance visualization of extradural compressive lesions, such as synovial cysts. A multitude of infectious agents are on the differential list for dogs with focal or multifocal neurological dysfunction. A number of Bartonella spp., including B. henselae, B. vinsonii subsp. berkhoffii, and B. clarridgeaie, are known to infect dogs. While multiple human cases of neurobartonellosis have recently been described, the role these organisms play in clinical diseases of the central nervous system of dogs has not been widely explored. The purpose of this study was to use polymerase chain reaction (PCR) to amplify Bartonella spp. DNA from cerebrospinal fluid (CSF) of naturally exposed dogs in endemic areas meeting criteria for inflammatory central nervous system (CNS) disease.

N19

Methods: Dogs with cytologically or histologically diagnosed tumors of any histology (excluding mast cell tumors) and any stage were allowed. A standard, open-label, phase I, 3 + 3 dosecohort escalation design was employed. The toceranib dose remained constant at 2.75 mg/kg administered orally every other day. Doxorubicin was initiated at 20 mg/m2 administered IV every 21 days for four doses. The doxorubicin dose was escalated by 5 mg/m2 each cohort until MTD or 30 mg/m2 was reached. DLT was defined as any grade 3 adverse event with the exception of hematopoietic AEs for which a grade 4 AE was considered dose limiting. Any grade AE that was refractory to supportive care or persisted beyond seven days was also considered dose-limiting.
T-test and Chi-square test were used to compare and match continuous variables (age, weight) and categorical variables (sex), respectively, between lymphoma and control groups. The cytokine concentrations were measured using a commercial canine multiplex magnetic bead-based (Luminex)Ò assay which measured IL-2, IL-6, IL-7, IL-8, IL-10, IL-15, IL-18, GM-CSF, TNF-a, IFN-c, IP-10, KC-like, and MCP-1. The serum levels of each cytokine were compared amongst the three groups (B cell lymphoma, T cell lymphoma, healthy control) using ANOVA or Kruskal-Wallis tests, as appropriate. Statistical significance was set at P < 0.05.
There was no significant difference between the lymphoma and healthy control groups regarding sex, age and weight. IL-6, IL-10, KC-like, and MCP-1 were significantly higher in dogs with lymphoma compared to healthy dogs (P = 0.01, P = 0.03, P = 0.01, P < 0.01, respectively). IL-10, KC-like, and MCP-1 were significantly higher in the B cell lymphoma group than in the healthy group (P = 0.01, P < 0.01, P = 0.01, respectively). MCP-1, IL-6 and IL-15 levels were significantly higher in the T cell lymphoma group than in the healthy group (P = 0.02, P < 0.01, P < 0.01, respectively). Finally, IL-6 and IL-15 were significantly higher in the T cell lymphoma group than in the B cell lymphoma group (P = 0.03. P = 0.02, respectively). There were no significant differences in serum IL-2, IL-7, IL-8, IL-18, GM-CSF, TNF-a, IFN-c, and IP-10 between the lymphoma and control groups.

O06

This study represents the first description of Bcl11b expression in the cat and in FOSCC. We demonstrated activation of the Src pathway by FOSCC cells making Src a strategic therapeutic target that could be used in a clinical setting. Exposing FOSCC cells to dasatinib has inhibitory effects on cell proliferation, EGFR, and consistently decreased production of VEGF. Based on these results, dasatinib may have both anti-proliferative and anti-angiogenic effects in FOSCC. Since EGF is able to partially rescue cells from the effects of dasatinib, future studies should evaluate dual target inhibition of Src and EGFR, EGFR and VEGFR, as well as combining Src inhibition with standard chemotherapy agents. Oral melanoma (OM) has a high propensity to metastasize to regional lymph nodes (LN) and lungs. Surgery and/or radiotherapy (RT) are effective local treatments, however most dogs succumb to distant metastasis. Immunotherapy represents an attractive strategy for this potentially immunogenic tumor. The objective of this multi-institutional retrospective study was to examine the clinical outcome of dogs with OM treated with ONCEPT TM melanoma vaccine, +/surgery and/or RT.
One hundred and twenty six dogs were included. All received ONCEPT TM . Sixty had adequate local control (ALC; complete excision or irradiation of microscopic disease). Fifteen were treated in the microscopic setting. Fifty-one were treated in the gross disease setting, of which 39 underwent RT. Median time to progression, median progression-free survival and median overall survival were 304, 244 and 617 days, respectively. Dogs with ALC had improved clinical outcomes. The following also correlated with favorable clinical outcomes: rostral location, stage 1, absence of LN metastasis, low mitotic index, absence of bony lysis, absence of gross disease. This is the largest reported series of dogs with OM treated with ONCEPT TM . Clinical outcomes are similar to those reported recently for dogs treated with surgery and/or RT alone. Several prognostic indicators were confirmed. A prospective, randomized, controlled study is needed to determine the clinical benefit of ONCEPT TM .

O08

ENHANCEMENT OF DOXORUBICIN EFFECTIVENESS WHEN COMBINED WITH SALINOMYCIN IN FISS CELL LINES. Lucia Borlle 1 , Brittany Zumbo 2 , Kelly R. Hume 1 . 1 Cornell University, Ithaca, NY, USA, 2 Michigan State University, MI, USA Feline injection site sarcoma (FISS) is an aggressive neoplasia that remains a challenge for clinicians. Radical surgery is the first choice treatment but is not always feasible. Chemotherapeutic responses have been documented but are generally of short duration; tumors often develop resistance quickly. Salinomycin is an ionophore that has been shown to inhibit cancer stem cells and to increase the sensitivity of human soft tissue sarcoma cells to doxorubicin chemotherapy. The aim of our study was to determine if salinomycin could be used to increase the sensitivity of FISS cell lines to doxorubicin. Two primary cell lines cultured from feline patients with histologically confirmed FISS were used for this purpose. Since the threshold response to doxorubicin differs, we ranked the cells according to the sensitivity to doxorubicin and used one sensitive line (C10) and one resistant line (B4). Cell viability assays (MTT) were used to determine the response to single agent and combination therapy. Three independent assays were performed, with samples evaluated in triplicate in each assay. Student's t-tests were employed to compare the results of individual doxorubicin concentrations with and without salinomycin combination therapy. Addition of 5 uM salinomycin resulted in a statistically significant (P < 0.05) reduction in cell viability compared to single agent doxorubicin at multiple concentrations for both cell lines. Our results highlight a potential chemotherapy protocol for the treatment of FISS. Further investigations need to be pursued to define the effect of this novel therapeutic combination and to validate our results in vivo. Canine Non-Hodgkin's lymphoma (NHL) occurs in approximately 30-100/100,000 dogs per year and accounts for up to 83% of canine hematopoietic cancers. Standard-of-care treatment for canine NHL comprises multi-drug chemotherapeutic protocols. 60-85% of dogs enter a complete clinical remission following induction chemotherapy, and median first-remission durations range from 140 to 385 days. Resistance to chemotherapy increases with each subsequent relapse and less than 10% of patients survive longer than 2 years from diagnosis. To prolong overall survival, more effective therapies are required to prevent relapse following successful induction chemotherapy. Directing the immune system to specifically target malignant cells is a powerful strategy in the treatment of cancer. We have previously developed a vaccine strategy that utilizes autologous CD40-activated B cells as antigen-presenting cells electroporated with autologous tumor RNA as the antigenic payload to stimulate anti-tumor immunity in dogs with NHL. In our first clinical trial, we showed that dogs vaccinated three times at three week intervals following successful induction chemotherapy had a tendency towards prolonged remission times and overall survival when compared to historical controls. Furthermore, vaccinated dogs that did relapse showed a statistically significant increase in overall survival following rescue chemotherapy. These results coupled with immune analysis from vaccinated patients suggested that while RNA loaded CD40-activated B cells primed an anti-tumor immune response, it was insufficient to maintain remission. Furthermore, we hypothesized that rescue chemotherapy boosted vaccine primed tumor-specific immunity and lead to prolonged second remission times. We therefore hypothesized that repeat administrations of RNA-loaded CD40-activated B cells given every 2 months following an initial priming course would maintain anti-tumor immunity and prevent relapse.

O10

The American College of Veterinary Internal Medicine Oncology and Small Animal Internal Medicine listserves were used to solicit data pertaining to cases in which toceranib was used in the treatment of feline mast cell neoplasia. Cases were included if the following data were received: signalment (age, gender, breed), anatomic classification of disease (cutaneous or visceral), previous and concurrent treatment, toceranib dose (mg/kg) and schedule, duration of therapy, best response, and documentation of adverse events.
Toceranib appears to be well-tolerated in feline patients with mast cell neoplasia. Biologic activity of this drug is evident in the studied cats, however further prospective studies are needed to fully elucidate its role in treatment of this condition. Gastrointestinal is the most common anatomical form of lymphoma in cats and systemic chemotherapy is indicated for the treatment. Although most of cats can achieve a complete remission with chemotherapy, relapse can occurs and may be associated to the multidrug resistance (MDR). Cell mechanisms of MDR include activation of transmembrane proteins, that reduce intracellular concentrations of different chemical compounds, and alterations in drug target. The main protein related to MDR phenotype is P-glycoprotein. Its overexpression results in reduced concentration of certain drugs within the cell, which is linked to the resistance. The aim of this study was to investigate P glycoprotein (P-gp) expression in cats with T-cell lymphocytic gastrointestinal lymphoma. Immunohistochemistry was performed in 40 samples using the monoclonal antibodies mouse anti-PAX-5 (PAX5-Invitrogen), polyclonal rabbit anti-CD3 (DAKO) and mouse anti-P-glycoprotein (clone C494-Enzo Life Sciences). P-gp immunoreactivity was analysed by degree of staining intensity, between 0 (no expression) to 3 (strong expression). Lymphoma was considered to be positive for P-gp when more than 10% of the neoplastic population expressed the protein. Three cats had a strong and 7 had a moderate expression, comprising 25% of the cases. The median of overall survival time was 24 months for these 10 cats; 60% had complete and 40% partial response to the treatment, and it was the same median observed for all cats studied. In conclusion, the expression of P-gp was observed in 25% of the cats with T-cell lymphocytic gastrointestinal lymphoma and this expression was not related to the treatment response or overall survival time. The purpose of this study was to describe initial experiences with a stereotactic radiation therapy (SRT) protocol (10 Gy x 3 daily fractions delivered with 6 MV photons via a linear accelerator) used as treatment for non-lymphomatous nasal tumors in dogs. A retrospective analysis of cases treated from August 2013 to October 2015 at NC State Veterinary Hospital was performed. Dogs were included if they had biopsy-confirmed nonlymphomatous nasal tumors. Dogs treated with chemotherapy were excluded. The gross tumor volume (GTV) was contoured using pre-and post-contrast 1-2 mm slice thickness CT image sets.
Twenty-five cases were included. Stages included T1 (n = 1), T2 (n = 10), T3 (n = 2), T4 (n = 12). None had evidence of distant metastasis. Nineteen had regional lymph node aspirates and metastasis was present in one. Tumor types included adenocarcinomas (n = 15), chondrosarcomas (n = 5), transitional nasal carcinomas (n = 3), intranasal squamous cell carcinoma (n = 1), and neuroesthesioblastoma (n = 1). Clinical signs improved in all cases. Twelve cases had recheck CT scans 3-4 months post-SRT and partial or complete tumor response was seen in all cases. Minimal acute toxicity was detected. The only late effect noted to date is oronasal fistula development in one dog 4 months post-SRT (this was anticipated due to tumor invasion into the palate at diagnosis). The median disease progression-free survival time has not been reached, and 52% of dogs are progression-free at 1 year. Nine dogs are deceased (median survival time 375 days) and 40% are alive 600 days post SRT.
Three fraction SRT has been used to treat non-lymphomatous nasal tumors in 25 dogs with limited adverse events. Continued accrual and follow-up will be necessary to confirm low toxicity, further elucidate effects of stage on outcome, and characterize clinical efficacy. Osteosarcoma (OSA) is locally invasive and highly metastatic. We hypothesize that standard-of-care carboplatin chemotherapy followed by continued administration of toceranib phosphate (TP) will result in improved survival in dogs with OSA as compared to historical controls that did not receive TP. In addition, we hypothesize that TP will decrease circulating levels of pro-angiogenic MMP-9, a downstream factor and surrogate marker for VEGF receptor inhibition.
The lack of replication-competent virus in saliva, urine, feces or peripheral blood satisfied Canadian Food Inspection Agency (CFIA) concerns regarding potential shedding, and enabled initiation of a clinical trial utilizing Maraba virus to treat cats with mammary cancer. In this trial a heterologous prime:boost strategy targeting tumour antigens will be tested. Introduction: Canine hemangiosarcoma is a highly malignant tumor, which is associated with poor long-term survival due to the development of early and widespread metastatic disease. Currently, little is known regarding the biology of canine hemangiosarcoma, and the mechanisms accounting for the highly metastatic nature of the tumor are poorly understood. In humans and rodents, monocytes have been shown to play key roles in metastasis through promotion of tumor cell extravasation, seeding, growth, and angiogenesis, as well as suppression of anti-tumor immunity. However, there has been little investigation into the role of monocytes in canine tumor metastasis. Therefore, we investigated the potential role of monocyte infiltration in the regulation of tumor metastasis in dogs.

EN01 GLUCOSE

HOMEOSTASIS DETERIORATES MORE RAPIDLY WITH AGE IN BURMESE CATS COMPARED TO NON-BURMESE. Margareth Rose Lederer 1 , Jacquie Rand 2 , Nubia Lopes 2 , John Morton 3 , Nick Jonsson 1 . 1 University of Glasgow, Glasgow, Scotland, UK, 2 University of Queensland, Gatton, Qld, Australia, 3 Jemora Pty Ltd., Geelong, Vic., Australia Burmese cats are predisposed to diabetes mellitus in Australia, New Zealand and United Kingdom, and the disease is 3 to 4 times more prevalent in this breed compared to domestic cats. Mechanisms predisposing Burmese cats to diabetes remain unclear. This study compared hormonal and biochemical variables associated with glucose and lipid metabolism between mixed-age, lean to overweight, clinically healthy Burmese and non-Burmese cats. Burmese cats older than 3.5 years had higher fasting glucose compared to non-Burmese (P = 0.02), with 1.3 mmol/L higher at 3.5 years and 3.5 mmol/L higher at 10 years of age (P < 0.001), but not at 2 years of age. Mean and 2-hr glucose concentrations during a glucose tolerance test were higher in Burmese than non-Burmese cats (P < 0.006 to 0.031), indicating relative glucose intolerance. Lean Burmese cats had 4.8 lU/mL higher fasting insulin concentrations than non-Burmese, suggesting they are insulin resistant (P = 0.031). At time 10 minutes during the GTT, Burmese cats had lower insulin concentration compared to non-Burmese, but were higher at 120 minutes (P = 0.066 and P = 0.046). This is consistent with diminished first phase, and increased and prolonged second phase insulin release, a pattern of insulin secretion described in prediabetic, diabetic, obese humans and obese cats. Fasting triglyceride concentration was 0.2 mmol/L higher (P = 0.015) and fasting NEFA concentration was 0.18 mmol/L higher (P = 0.016) in Burmese compared to non-Burmese. These results demonstrate that Burmese cats have more rapid deterioration of b-cell function with age, and impaired glucose homeostasis and lipid metabolism occur concurrently in these cats. Previous studies indicated that serum fructosamine level is decreased in hyperthyroid cats; however, its clinical utility in diabetic hyperthyroid cats was not reported. We hypothesized that hyperthyroidism in diabetic cats will result in a clinically significant decrease in serum fructosamine level compared to euthyroid diabetic cats. Data of serum total thyroxine (TT4) and fructosamine of hyperthyroid/euthyroid diabetic/nondiabetic cats from the New Zealand Veterinary Pathology and Colorado State University were retrieved and statistically analyzed. Serum fructosamine was significantly lower in hyperthyroid diabetic than euthyroid diabetic cats (mean 332 µmol/L 95% CI 291-379, n = 18 versus mean 527 µmol/L 95% CI 515-553, n = 186) while not different between hyperthyroid diabetic and euthyroid nondiabetic cats (mean 332 µmol/L 95% CI 291-379, n = 18 versus mean 321 µmol/L 95% CI 296-345, n = 128). There was a significant negative correlation between serum TT4 concentration and serum fructosamine and glucose concentrations (n = 659, P < 0.01, and n = 297, P < 0.01, respectively). Hyperthyroid cats (diabetic or not) had significantly (P < 0.05) lower serum glucose than euthyroid cats (diabetic or not). Diabetes and random variability accounted for 49.7% and 45.4% of the change in serum fructosamine; whereas, age (3.1%) and population (1.8%) had minimal impact on serum fructosamine. In conclusion, in the current study, serum fructosamine concentration was significantly decreased in diabetic hyperthyroid cats and was similar to the fructosamine concentration in euthyroid nondiabetic cats. In addition, approximately 45% of the change in serum fructosamine was independent of diabetes. Future studies should be undertaken to explore specific factors that have an impact on fructosamine level. Insulin aspart (Aspart) is a rapid-acting insulin analog that has been shown to be an effective treatment for diabetic ketoacidosis (DKA) when administered by intermittent subcutaneous (SC) injection in humans. In cats, rapid-acting insulin analogs have not yet been evaluated, and the most commonly used protocols for DKA involve delivery of regular insulin via constant intravenous infusion, which can be technically challenging. The aim of this study was to determine the pharmacodynamics (PD) and pharmacokinetics (PK) of Aspart following SC and intramuscular (IM) injections in healthy cats.

EN03

Aspart is rapidly absorbed following SC and IM injection in healthy cats. The effective and predictable glucose lowering effect of insulin aspart may be advantageous in the treatment of DKA in cats. Obesity is a major risk factor for the development of diabetes in cats. Recently, retinoid-related orphan receptor gamma (RORc) has been identified as an important transcription factor in the development of large insulin-resistant adipocytes. RORc can be inhibited by its ligand tetra-hydroxylated bile acid (THBA). Oral supplementation of THBA reduces adipocyte size, improves insulin sensitivity and prevents hyperglycemia in obese mice. The purposes of this study were to investigate palatability and possible side effects of THBA, and the effects of THBA supplementation on adipocyte size and insulin sensitivity in healthy cats.

EN04

Six healthy purpose-bred cats were fed 5 mg/kg/day of THBA for 8 weeks. THBA was given in a small amount of food to each cat individually before the daily ration was offered. Complete blood count, biochemical profile and insulin tolerance test (ITT; 0.1 IU/kg insulin aspart IV) were performed at weeks 0 and 8. After 12-18 hours of fasting blood glucose concentration was recorded and the rate constant for the disappearance of glucose (K ITT ) was calculated. Subcutaneous fat tissue samples were taken in the umbilical area by a 4 mm biopsy punch, and adipocyte size was determined by histological and image based analysis of at least 10 sections per animal. Data were analyzed by parametric tests (P < 0.05).
Oral supplementation of THBA was well accepted in all cats, and no side effects were noted. None of the cats displayed abnormalities on complete blood count and serum biochemical profile. Fasting blood glucose concentration and average size of white subcutaneous adipocytes were significantly lower after 8 weeks of THBA administration, whereas K ITT did not differ.
This retrospective study of cats aged ≥10 years, referred to a single UK veterinary teaching hospital (2006) (2007) (2008) (2009) (2010) (2011) (2012) (2013) (2014) , used electronic patient records to obtain owner-reported data regarding breed, coat colour, age and sex, and clinical data to classify cats as hyperthyroid/euthyroid. Bayesian multivariable logistic regression was used to evaluate breed, age and sex as risk factors for hyperthyroid status in all the cats (purebred and non-purebred). To avoid interactions with coat length or colour in purebreds, the effect of coat colour/pattern, colour dilution, base pigment, white markings and hair length were assessed only in non-purebred cats in a separate analysis. Variables with P < 0.2 in univariate analyses were evaluated in multivariable models, with final variables significant at P ≤ 0.05. Risk factors are quantified as odds ratio ((OR) P-value).
Of 3934 cats included in the final analysis, 885 were hyperthyroid and 3049 euthyroid. Multivariable results showed Burmese (OR 0.01, P = 0.005), Persians (OR 0.20, P < 0.001), Siamese (OR 0.32, P = 0.006), Abyssinians (OR 0.04, P = 0.035) and British shorthairs (OR 0.52, P = 0.014) had reduced risk of hyperthyroidism compared to domestic shorthairs. Domestic longhairs (OR 1.34, P = 0.015) showed increased risk of hyperthyroidism compared to domestic shorthairs, but coat colour, colour dilution, base pigment and white markings did not have a significant effect. Overall, females (OR 1.39, P < 0.001) had increased risk of hyperthyroidism compared to males, and, compared to 10 year olds, increased risk was found in cats aged 11-12 years (OR 1.45, P = 0.007), 13-14 years (1.83, P < 0.001) and 15-17 years (OR 1.50, P = 0.004).
To authors' knowledge this is the first study to report increased risk of hyperthyroidism in long haired, non-purebred, domestic cats. Consistent with a number of previous studies, increased risk of hyperthyroidism was found with increased age and female sex, and reduced risk in Burmese, Siamese and Persian breeds. However, this study also newly identified two further purebreds, Abyssinian and British shorthair, at reduced risk of hyperthyroidism, neither of which were exclusively colourpoint. Coat colouration was not found to be associated with risk of hyperthyroidism in the analysis of domestic cats, however, reliance on secondary data may have resulted in misclassification errors and coat phenotype may be a poor surrogate marker for melanin concentrations. Further studies are required to evaluate tyrosine availability as a potential factor in the aetiology of feline hyperthyroidism, and to investigate the apparent protective effect in certain purebreds and associations between hyperthyroidism and hair length. Medical records were searched for cats receiving 131-I therapy after documented feeding of the prescription diet for at least 14 days. Cats were excluded if the prescription diet had been supplemented with other foods, or if methimazole had been administered concurrently with, or subsequent to, the prescription diet. Owners were contacted directly to provide missing dietary information.
Mean serum concentrations of T 4 and fT 4 were significantly higher and TSH was significantly lower during levothyroxine administration compared to baseline in both groups. Mean serum concentrations of T 4 , fT 4 and TSH beginning 1 week after levothyroxine was discontinued were significantly different compared to values during levothyroxine administration but not compared to baseline values. When groups 1 and 2 were compared, there was no difference in mean T 4 , fT 4 , and T 3 concentrations between groups during or after levothyroxine supplementation. Suppression of the HPTA occurred during levothyroxine supplementation, with mean serum T 4 , fT 4 and TSH concentrations returning to the reference interval by 1 week after discontinuation in both groups. It appears that assessing thyroid function tests 1 week after longterm levothyroxine supplementation may reliably establish dogs as being euthyroid. Autoantibodies directed against the P450 side chain cleavage enzyme (P450scc) have been recently described in a proportion of dogs affected with hypoadrenocorticism, consistent with an immune-mediated disease process. In humans affected with Addison's disease, autoantibodies can have a predictive value, being detected prior to clinical signs developing, and have been shown to persist post-diagnosis. Furthermore, an autoantibody positive status post-diagnosis has been associated with successful remission of Addison's disease following B-cell depletion with rituximab, suggesting active pathology in these cases.
This study demonstrates persistence of P450scc autoantibodies in a subset of dogs affected with hypoadrenocorticism and seroconversion over one year post-diagnosis. Whilst P450scc autoantibody positivity has been associated with sex, with females having a higher prevalence, there was no sex difference in persistence demonstrated. P450scc expression in the ovary may act as an additional source of antigenic stimulation in female dogs, explaining the later seroconversion observed in one individual. However, paralleling humans with Addison's disease, antibody persistence in dogs with hypoadrenocorticism might represent persistent pathology, due to residual antigenic stimulation and autoimmune inflammation in the adrenal gland. While hypoadrenocorticism, or Addison's disease, can occur in a dog of any breed at any age, certain breeds, including Nova Scotia Duck Tolling Retrievers (NSDTRs), have an increased incidence of the disease. Specifically in the NSDTR, there appears to be at least two forms of Addison's disease in the breed: a juvenile onset form (JADD) occurring in dogs under 1 year of age, and an adult onset form occurring in dogs at 4.5 years of age on average. JADD differs from adult onset Addison's disease, as it can be a multisystemic illness and appears to involve an autoimmune component. This is supported by the identification of a CD3+ T cell infiltrate in histologic sections of a JADD affected adrenal gland sample and that many cases are often affected with other autoimmune diseases, including immune-mediated hemolytic anemia and thrombocytopenia, immune-mediated polyarthritis, and hypothyroidism. To identify a genetic basis of JADD, a genome-wide association study was performed using the Illumina Canine HD 173,000 SNP array with 14 NSDTRs diagnosed with Addison's disease less than 1 year of age and 33 healthy control NSDTRs over 6 years of age. All cases were definitively diagnosed through an adrenocorticotrophic hormone stimulation test. Genome-wide association analysis identified a 1.7 Mb associated haplotype on chromosome 27. Whole genome sequencing was performed on 2 NSDTRs with the associated haplotype, as well as 6 unaffected NSDTRs and 11 other dogs from 5 different breeds. Analysis of variants yielded 5 segregating SNPs in the associated region: 1 intergenic, 2 intronic, 1 coding, and 1 in the 3 0 untranslated region of a gene. The only coding variant, a missense mutation causing an amino acid change from a proline to a leucine, occurs in a currently uncharacterized gene, but is predicted to be damaging and deleterious based on sequence conservation. In summary, genome-wide association and whole genome sequencing analysis has identified a novel gene and mutation implicated in multi-organ autoimmunity and juvenile onset Addison's disease in the NSDTR. The adrenocorticotropic hormone stimulation test (AST), often used for the diagnosis of canine hyperadrenocorticism, has a wide range of sensitivity and specificity. Thus, we hypothesized that standardized negative feedback on the hypothalamic-pituitary-adrenalaxis will result in large variability in the post AST serum cortisol concentrations. Therefore, this study aimed to evaluate the variability in the post AST serum cortisol in healthy dogs and to statistically examine parameters that may influence cortisol responses.

EN10

Fourteen healthy Harrier Hound dogs (mean age 7.1yAE3.2; mean body weight 26.5 kgAE2.7) were randomized by a Latin square design to receive 5 treatments [placebo, 50 mg, 37.5 mg, 25 mg, and 12.5 mg of cortisone acetate (CA)] in each of the five 7-day treatment periods, which were separated by 14-day washout periods. An AST was performed 24 hours after the last dose of CA for each of the 5 treatments and serum cortisol was analyzed by an electrochemiluminescence immunoassay.
A significant dose dependent decrease in the mean (AESEM) post AST serum cortisol was observed (placebo 365 AE 13 nmol/L, CA 12.5 mg 347 AE 13 nmol/L, CA 25 mg 335 AE 13 nmol/L, CA 37.5 mg 326 AE 13 nmol/L, CA 50 mg 308 AE 13 nmol/L). There was a significant (P < 0.05) between-week variability in post AST serum cortisol in the placebo and 25 mg and 37.5 mg CA groups. The overall variability in the post AST serum cortisol concentrations was attributed to the individual dog (31%), random unexplained variability (31%), dose of CA (16%), age (15%), body weight (4%), and timing of treatment (3%).
Sixty-nine cases were identified. The most common diagnoses were IHC (48%) and chronic kidney disease (35%). Urolithiasis was diagnosed in 14% (43% calcium oxalate), neoplasia in 13% (46% lymphoma) and primary hyperparathyroidism in 3%. Other diagnoses included hyperthyroidism (8%), hypertrophic cardiomyopathy (4%), feline idiopathic cystitis (2%), and diabetes mellitus (2%). The average age at diagnosis of hypercalcemia was 10.0 AE 4.61 years. Most common clinical signs reported in the IHC group versus other diagnoses (in which clinical signs could be related to the primary disease, and not necessarily to hypercalcemia), were anorexia (21.7% versus 31.9%), vomiting (14.5% versus 26.1%), lethargy (10.1% versus 7.2%), and constipation (10.1% versus 1.4%).
Medical records for dogs in which an ACTH stimulation test had been performed for a clinical suspicion of hypoadrenocorticism between 2005 and 2015 were retrospectively reviewed. Dogs that had received treatment for hyperadrenocorticism or recent glucocorticoid therapy were excluded. Five hundred and fourteen dogs met inclusion criteria, including 163 dogs with hypoadrenocorticism (post-ACTH stimulated cortisol concentration ≤55 nmol/ L). An ROC curve was constructed for basal cortisol concentrations using commercially available software. The area under the ROC curve was 0.991. The ROC curves were nearly identical when evaluating dogs with and without electrolyte abnormalities. Accuracy was optimal at 22 nmol/L, which provided a sensitivity and specificity of 96.9% and 95.7%, respectively. At cut-offs of 55 nmol/L and 4 nmol/L, sensitivity and specificity were 99.4% and 99.7%, respectively. Using a previously estimated prevalence of 15%, cut points of 55 nmol/L and 4 nmol/L resulted in negative and positive predictive values of 99.8% and 97.9%, respectively. Even at lower prevalence rates, positive predictive values remained high at low cut points. Results of this study reaffirm the high sensitivity and negative predictive value of basal cortisol concentrations for the diagnosis of hypoadrenocorticism. However, the high specificity and positive predictive values at low cut points could be of major importance for veterinary practitioners. Prospective studies are needed to further substantiate these findings. Congenital hypothyroidism (CH) is associated with significant morbidity and mortality in pediatric felines. Thyroid hormones (TH) are essential for normal development of the nervous and skeletal systems. The paucity of information on TH levels and the inability to identify early histopathologic changes has led to CH underdiagnosis. The literature on TH levels in kittens is sparse and incongruent, comprised of mostly case reports. References state kitten total thyroxine (TT4) levels are 2 to 3 times higher than the adult cat TT4 level. They also make the assumption free thyroxine (fT4) levels are higher than the adult cat. An abstract was previously presented showing kitten TH levels did not exceed the adult cat normals. These values were measured with assays that are no longer used. The normal pediatric TH levels must be evaluated with the newer and currently available assays. The purpose of this study was to determine the TT4, fT4, total tri-idiothyronine (T3), free tri-idiothyronine (fT3) and thyroid stimulating hormone (TSH) levels in healthy kittens aged 2 through 16 weeks measuring levels at weekly intervals using the current available diagnostic methods.
The TH levels in pediatric kittens were measured with the newer assays and with the exception of TT4, all TH levels were within the ACRR. The TT4 did not elevate more than twice the upper limit of the ACRR at any time point and was consistently within the ACRR after 12 weeks of age. A peak in TT4, fT4, TT3 and fT3 was seen at 6 weeks and a peak in TT4 and fT4 was also seen at 10 weeks. TSH levels using the canine TSH assay have been validated for use in adult cats, but these values have not yet been reported in kittens. The TSH levels remained within the ACRR and it showed a similar peak at 10 weeks. These patterns likely occur due to an increased growth and development at 6 and 10 weeks. Conflicting results have been reported regarding lipid concentrations in dogs with diabetes mellitus (DM). Some studies have reported that low-density lipoprotein-cholesterol (LDL-C) is not significantly different in dogs with and without DM. The goal of this study was to report total cholesterol (TC), total triglycerides (TG), high-density lipoprotein-cholesterol (HDL-C), very lowdensity lipoprotein-cholesterol (VLDL-C), and LDL-C in fasted dogs with and without DM.
Hydrocortisone administration resulted in increased abdominal visceral fat mass on days 30, 60, and 90 compared with fat mass before administration (day 0). Additionally, visceral fat area at the L3 level also was increased during hydrocortisone administration. Serum leptin concentrations began to increase on day 1 of hydrocortisone administration and were significantly increased on days 1, 3, 7, 30, 60, and 90 compared with concentrations on day 0. Serum adiponectin concentrations on days 1, 3, 7, 30, 60, and 90 were decreased compared with concentrations on day 0. The mRNA expressions of leptin and adiponectin in the abdominal fat were increased on day 30 compared with expressions on day 0, but the expressions were decreased on days 60 and 90 compared with those on day 30. We identified a correlation between serum leptin and adiponectin concentrations and visceral fat distribution. This study showed that hydrocortisone administration affected visceral fat distribution and serum concentrations of leptin and adiponectin by dysregulating leptin and adiponectin expression. Recombinant DNA technology has been used to modify the structure of human insulin to yield analogs that are more rapidlyacting than regular insulin or have a longer duration of action than NPH insulin. The long-acting insulin analogs have been studied in cats and are routinely used to manage feline diabetics. More recently, the isoglycemic clamp method has been used to study the metabolic effect of a rapid-acting insulin analog in healthy cats. Generating time-action profiles to study the pharmacodynamic properties of insulin formulations does not require measurement of plasma insulin concentrations, but if measured, they provide useful pharmacokinetic data that can be used to validate the timeaction profiles. The Iso-Insulin ELISA (Mercodia AB, Uppsala, Sweden) is used to measure the concentration of insulin analogs in plasma. A previous study using actual feline serum samples with a wide range of insulin concentrations measured by a commercial ELISA failed to detect cross-reactivity of feline insulin with the Iso-Insulin ELISA.

EN18 STEROID HORMONE PANEL TEST AND TRILOSTANE AND MELATONIN THERAPY IN POMERANIAN DOGS

Food was withheld 18 -24 hours, and a cephalic vein catheter implanted. A venous blood sample was collected and the serum and plasma separated and frozen at À80°C. A glucose tolerance test (glucose 0.5 g/kg) was performed. Biochemical (screening, fasting and 2-hr blood glucose in the glucose tolerance test, triglyceride), hormonal (leptin, adiponectin, leptin:adiponectin ratio, insulin, glucose:insulin ratio) and enzymatic (fPLI, MCP-1) analyses were performed on plasma and serum samples. After extraction, analyses using GC-MS was performed. Linear modeling was used to identify statistically significant (adjusted P-value<0.05) metabolites associated with obesity and whether these responses were affected by sex and age. Pearson correlations were used to identify relationships between metabolites and other parameters.
A total of 49,930 SNPs were available for analysis. After excluding cats with low genotypic rate, 389 control DSH and 192 lean diabetic DSH cats were evaluated. Diabetic cats had a mean (SD) age of 11.62 (3.44) years; 123 (63%) were male, 71 (37%) female. Nondiabetic cats had a mean (SD) age of 14.83 (2.06) years; 216 (54%) were female, 183 (46%) male. Control cats were significantly older than diabetic cats (P < 0.0001; t-test). Five significant SNPs were identified: chrA2.4150731 (P = 1.4 x 10 À7 ); chrUn17.115508 (P = 7 x 10 À7 ); chrUn17.394136 (P = 3 x 10 À7 ); chrUn17.314128 (P = 3 x 10 À7 ) and chrUn17.7283 (P = 9 x 10 À6 ). The first SNP is located within chromosome A2; the others are located within a 0.8 Mb region towards the end of chromosome A3. The SNP in chromosome A2 is located 3 kb upstream of dipeptidyl-peptidase-9 (DPP9), a peptidase similar to DPP-4, involved in incretin inactivation. Within the identified region of chromosome A3, genes of interest include TMEM18 and ACP1; both have been associated with T2DM in humans, most likely causing insulin resistance. This is the first GWAS of DM in cats. A number of significant SNPs have been identified, some of which are located in proximity to genes that have been associated with T2DM in humans; others could be involved in pathophysiology related to DM. Further investigation of these candidate genes is warranted.
Disclosures: Disclosures to report. SNP chips for the GWAS were provided by the Morris Animal Foundation. Elevated serum canine pancreatic lipase immunoreactivity (cPLI) concentrations are highly specific for pancreatitis. Additionally, some studies in dogs with primary gastrointestinal (GI) disease have suggested a negative outcome in dogs with increased serum cPLI concentrations. GI disease, including foreign bodies (FBs) can mimic clinical signs of pancreatitis. GI FBs have also been proposed as a risk factor for the development of pancreatitis in dogs. To date the prevalence of pancreatitis in dogs with GI FBs is unknown. The primary aim of this study was to determine the prevalence of elevated cPLI concentrations (as measured by Spec cPL Ò ) in dogs with GI FBs, and to correlate Spec cPL concentrations with patient survival. A secondary aim was to compare Spec cPL concentrations among dogs based on age, FB type, FB location, and type of removal procedure.

GI01

Serum Spec cPL was measured in 49 dogs prospectively enrolled with endoscopically or surgically confirmed GI FBs from May 2014 to May 2015. Samples were obtained the day of the procedure prior to anesthetic induction. Dogs were retrospectively grouped based on age (≤4 years, n = 22; 5-8 years, n = 11; ≥9 years, n = 16), FB type (linear, n = 14; discrete, n = 35), FB location (stomach, n = 21; duodenum, n = 3; jejunum, n = 12; involving multiple segments, n = 13) and type of removal procedure (endoscopy, n = 19; surgery, n = 30). The prevalence of a serum Spec cPL above the upper limit of the reference interval (≥ 200 µg/L) or the suggested cut-off value for pancreatitis (≥ 400 µg/ L) was determined in all dogs. Serum Spec cPL concentrations were compared amongst groups using Kruskal-Wallis or Mann-Whitney U tests, as appropriate. Statistical significance was set at P < 0.05.
A Spec cPL≥ 200 µg/L was identified in 12/49 dogs (24.5%), 9 of which (18.4%) had a Spec cPL≥ 400 µg/L. Spec cPL (median, range) was higher in dogs 5-8 years (125 ug/L, <30-2885 µg/L) and ≥9 years (233 µg/L, 43-2816 µg/L) of age than in dogs ≤4 years (59 µg/L, <30-246 µg/L; P < 0.0001) of age. Spec cPL was higher in dogs with linear FBs (376 µg/L, <30-2885 µg/L) than in dogs with discrete FBs (68 µg/L, <30-837 µg/L; P = 0.0038). Spec cPL was higher in dogs with FBs affecting multiple GI segments (569 µg/L, <30-2885 µg/L) than in dogs with gastric FBs (60 µg/L, <30-185 µg/L; P = 0.0159). Spec cPL was higher in dogs undergoing surgery (108.5 µg/L, <30-2885 µg/L) than in dogs undergoing endoscopy (63 µg/L, <30-185 µg/L; P = 0.0322). Two of 49 dogs (4%) were euthanized during the course of the study. Both dogs had evidence of septic peritonitis at the time of euthanasia. In both dogs, Spec cPL concentrations were elevated (1292 µg/L and 2816 µg/L, respectively).
The overall prevalence of Spec cPL ≥ 200 µg/L and ≥ 400 µg/L in dogs with GI FBs was relatively low (24.5% and 18.4%, respectively). Spec cPL was higher in middle aged and older dogs, dogs with linear FBs, and dogs that needed to undergo surgery for FB removal. A Spec cPL≥ 400 µg/L was noted in both dogs that were euthanized. Due to this small number, additional studies would be needed to confirm if an elevated Spec cPL is associated with a negative outcome in dogs with GI FBs. Prednisone and other glucocorticoids (GCs) are commonly used as long-term medications to treat a multitude of immune mediated, inflammatory, and neoplastic diseases in dogs. Patients with many of these diseases may have clinical signs that overlap with those of patients with pancreatitis. The effect of long-term GC administration on canine serum Spec cPL concentrations has not been adequately assessed. Therefore, the aim of this study was to determine if long-term administration of GCs at supraphysiologic doses affects serum concentrations of Spec cPL in dogs with a variety of different diseases.
Dogs were prospectively enrolled at the Veterinary Specialty Hospital of San Diego between February 2015 and October 2015. Inclusion criteria for enrollment included prescribed long-term (≥ 3 weeks) administration of at least 0.5 mg/kg/day prednisoneequivalent of GCs. Serum Spec cPL concentration was measured prior to initiation of GC therapy so each dog served as its own control. A second measurement of serum Spec cPL concentration was obtained in all dogs after at least 3 weeks of treatment with GCs. Only dogs that had no dosage change within one week of sampling were evaluated. The dose of GCs recorded was the dose that was being administered at the time that the second serum sample was collected. Cases were excluded if there was poor owner compliance with GC administration (defined as missing more than two consecutive doses of GCs) or if there was administration of other medications known or suspected to increase serum Spec cPL concentrations (e.g. potassium bromide or phenobarbital) or GC administration within the 3 months prior to enrollment. Comparison of serum Spec cPL between pre GC administration and ≥ 3 weeks post GC administration was performed using Wilcoxon matched-pairs signed rank test. Statistical significance was set as P < 0.05.
One hundred and seven cases (5 cats and 102 dogs) with a total of 111 fishhooks were included. Single shanked hooks comprised 52/111 (46.85%) and multiple shanked hooks comprised 59/111 (53.15%) of cases. Most common locations were the pharyngeal/ oral region 38/111 (34.26%)), cervical esophagus (25/111 (22.52%)), and stomach (27/111 (24.32 %)). Three hooks were cutaneously embedded. Hooks with multiple shanks were 2.314 times more likely to be associated with esophageal damage than single shanked hooks (95% CI: 1.038-5.159; P = 0.0478). Endoscopic retrieval was successful in 60/69 (87.0%) cases in which it was attempted. Esophageal perforation occurred in 2 endoscopic cases and a total of 9 cases necessitated surgery. Survival was 100%.
Our study supports that endoscopic retrieval is highly successful and survival is 100%, even in cases requiring surgery. While multiple shanked fishhooks are more likely to cause esophageal damage, neither type of hook nor location affected outcome.

GI06

Predominant phyla throughout the samples were Proteobacteria, Firmicutes, Fusobacteria, and Bacteroidetes. Percentages of reads identified as Proteobacteria decreased aborally (P = 0.017; duodenum, 62%; ileum, 48%; colon, 9%; rectum, 6%), while those of Firmicutes increased (P = 0.032; 20%, 43%, 68%, 69%). On the gas chromatography platform, 530 unique compounds were detected, and 199 of these were assigned to named metabolites. 134 named metabolites had at least one site-pair comparison reaching statistical significance. Key fermentation compounds such as sugars and amino acids had varied site distributions: glucose increased progressively along the GIT along with 6-deoxyglucose (P = 0.047 and 0.034, respectively). Nearly all amino acids exhibited a decrease between the ileum and colon (e.g., alanine, citrulline, glutamine, glycine, lysine, methionine, tryptophan, and valine, all P < 0.03).

GI09

To be included into the study, the dogs had to be at least 1 year of age, have clinical signs of EPI, a serum cTLI concentration ≤2.5 lg/L, and be free from any other concurrent disease. Three naturally voided fecal samples collected over three consecutive days were frozen immediately after collection and pooled. Fecal samples were collected in a similar manner from healthy dogs without any clinical signs of gastrointestinal disease. Extracted DNA from fecal samples was used for Illumina sequencing of the bacterial 16S rRNA gene and analyzed using Quantitative Insights Into Microbial Ecology (QIIME). The analysis of similarities (ANOSIM) function in the statistical software package PRIMER 6 (PRIMER-E Ltd., Luton, UK) was used on the unweighted UniFrac distance matrix to determine if any groups of samples contained significantly different bacterial communities. There was a significant difference in fecal microbial communities when healthy dogs were compared to treated (P = 0.001) and untreated (P = 0.001) dogs with EPI. Quantitative Insights Into Microbial Ecology (QIIME). The analysis of similarities (ANOSIM) function in the statistical software package PRIMER 6 (PRIMER-E Ltd., Luton, UK) was used on the unweighted UniFrac distance matrix to determine if any groups of samples contained significantly different bacterial communities. There was a significant difference in fecal microbial communities when healthy dogs were compared to treated (P = 0.001) and untreated (P = 0.001) dogs with EPI. Alpha diversity was significantly decreased in untreated and treated EPI dogs when compared to the healthy dogs (P < 0.01). The families Bifidobacteriaceae (P = 0.006), Enterococcaceae (P = 0.035), and Lactobacillaceae (P = 0.001) were significantly increased in the untreated and treated dogs with EPI when compared to healthy dogs. In contrast, Lachnospiraceae (P < 0.001), and Blautia (P < 0.001) were significantly decreased in dogs with EPI.
Eight cats were used in a repeated-measures study. Cats were fed a maintenance diet (M-diet) from study days -21-0 then switched to a weight-loss diet (WL-diet) on day 0 without calorie restriction until day 7. On days 8-84, daily intake was restricted to achieve 1-2% weight-loss per week. On days 84-98, feeding the M-diet resumed while maintaining stable body condition score (BCS). Fecal samples were collected and stored at À80°C until analysis. Samples were collected on day À7 and 0 (Obese/M-diet), 5 and 7 (Obese/WL-diet), 77 and 84 (Lean/WL-diet) and 86, 89, 91 and 98 (Lean/M-diet) for microbiota and SCFA profiles. Body weight, BCS, muscle condition score, and body fat percentage were assessed throughout the study.
Microbiota and SCFA profile alterations were noted based on body condition. Further study is warranted to determine the impact of these changes on inflammation and metabolic disorders. Canine inflammatory bowel disease (IBD) is an immunemediated enteropathy likely triggered by environmental and immunoregulatory factors in genetically susceptible dogs. Previous studies suggest a pivotal role for intestinal bacteria in disease pathogenesis since luminal microbial composition is markedly altered at diagnosis. It has been suggested that probiotic bacteria are effective and promising agents for the treatment of IBD in humans and animals. However, the effects of probiotic bacteria that directly interact with the host's resident intestinal microbiota are poorly understood. The aim of the present study was to investigate the impact of multi-strain probiotic VSL#3 supplementation (Visbiome; Exegi Pharma) at a dose of 50 billion bacteria/kg/day on the fecal and intestinal mucosal microbiota in dogs with IBD. Nineteen dogs diagnosed with moderate-to-severe IBD (CIBDAI ≥ 6) were randomized to receive standard IBD therapy (ie, elimination diet and prednisone) with or without probiotic VSL#3 for 8 weeks. Dogs were evaluated clinically at 0, 3, and 8 weeks and endoscopically and histopathologically at 0 and 8 weeks. The fecal and mucosal microbiota were investigated before and during treatment using 16S rRNA based quantitative polymerase chain reaction (qPCR) and fluorescence in situ hybridization (FISH), respectively. Both cohorts of IBD dogs showed a reduction in GI signs after 8 weeks of therapy compared to baseline CIBDAI scores (P < 0.05). Using qPCR, the placebo group failed to show significant changes in any of the bacterial groups analyzed. In contrast, VSL#3 treated dogs showed a significant (P = 0.001) increase in total lactic acid bacteria (Bifidobacteria, Streptococci, Lactobacilli) at treatment weeks 3 and 8 versus baseline values. Bifidobacteria were also significantly (P = 0.018) increased at 3 and 8 weeks post-VSL#3 treatment. FISH analysis showed that VSL#3 altered the number and spatial distribution of most colonic mucosal bacterial groups including total bacteria and the number of Bifidobacteria, Fecalibacteria, Lactobacilli, Streptococci, and Enterobacteriaceae (P < 0.05 for each). The number of mucusladen and attaching Bifidobacteria and Streptococci were also significantly (P < 0.05) increased in dogs receiving VSL#3. In conclusion, probiotic therapy of IBD dogs with VSL#3 increases the number of fecal and mucosal probiotic genera, especially the lactic acid bacteria. IBD dogs receiving standard therapy (placebo) showed little change in the composition of microbial communities despite exhibiting clinical improvement. Chronic enteropathy (CE) is a multi-factorial disease, which involves aberrant immune responses to commensal bacteria or dietary antigens. Although macrophages have been shown to play an important role in human disease very little information is available in dogs. CD163 is a scavenger receptor specific for macrophages, and has been detected in canine duodenum. The aim of this study was to determine if intestinal CD163 positive cell (CD163 + ) populations alter after successful treatment of CE. Calprotectin was used as an additional marker for macrophages. Dogs were prospectively enrolled and underwent standard diagnosis and treatment trial for CE. Duodenal biopsies were taken before and after long-term resolution of clinical signs (≥2 months). Serial histologic sections were stained for CD163 (using AM-3K) and calprotectin. Positively stained cells in five villous and crypt regions were manually counted. The percentages of positively stained cells from the total nucleated cells per 10,000 lm 2 before and after treatment were compared with repeated-measure ANOVA.

GI13

These results suggest that macrophages may play a role in canine CE. Further characterization of macrophages is warranted to understand their role in CE. The term protein-losing enteropathy (PLE) is often applied to cases of chronic enteropathy (CE) when the disease process results in hypoalbuminemia. Multiple previous studies have identified hypoalbuminemia as a risk factor for a negative outcome in dogs with chronic enteropathy, yet it has not been determined whether the histopathology differs between CE dogs with and without hypoalbuminemia. The goal of this retrospective study was to compare histopathologic findings (as determined by the WSAVA scoring system) in dogs with CE with and without hypoalbuminemia.

GI14

Dogs with PLE secondary to CE had a greater percentage of lacteal dilation, injury to the villi, crypt distension, and lamina propria neutrophils than dogs with CE that had normal albumin.
In conclusion, the human granulocyte immunofluorescence assay used in this study was able to identify sero-positivity for ANCA in a high percentage of dogs with FRD. The data suggests that this assay could be used as an alternative to the canine assay for detection of ANCA in dogs. Citrulline, a non-proteinogenic amino acid, has recently been described as a biomarker of enterocyte function in humans. Decreased concentrations of plasma citrulline have been reported in patients with short bowel syndrome, Crohn's disease, viral enteritis, or in critically ill patients. In dogs, the relationship between serum citrulline concentration and small intestinal dysfunction has not been well characterized. Citrulline is mainly synthesized from glutamine in enterocytes and is metabolized in the duodenum and jejunum. Therefore, intestinal injury can lead to decreased serum citrulline concentrations. The aims of this study were to analytically validate a GC/MS method for the measurement of citrulline in canine serum and to assess the concentration of this amino acid in dogs with chronic enteropathy.

GI16

The assay evaluated here was shown to be linear, precise, accurate, and reproducible. Dogs with chronic enteropathy had significantly lower serum citrulline concentrations than healthy dogs. Further studies are warranted to assess the clinical utility of the serum citrulline as a biomarker for enterocyte function in dogs. Dogs with symptoms of CE and a serum cbl below 285 ng/L (reference interval: 244-959 ng/L) were included from Evidensia Specialist Animal Hospital, Helsingborg, Sweden, Helsinki University Hospital, Finland, and from two other Swedish Veterinary Clinics. Dogs were randomized to treatment with either daily PO cbl tablets during the whole study period (cyanocobalamin (Behepan) 1 mg/tablet; dogs up to 10 kg received ¼ tablet, dogs 10 to 20 kg ½ tablet and dogs ≥ 20 kg 1 tablet/day) or PE cbl according to a protocol currently suggested by a diagnostic laboratory (http://vetmed.tamu.edu/gilab/research/cobalamin-information#dosing). A block-randomized schedule was performed by an external statistician prior to the start of the study in March 2014. The study was approved by the Swedish Board of Agriculture and the Viikki Campus Research Ethics Committee, Finland. Serum cbl for follow-up was analyzed 28 AE 5 days and 90 AE 15 days after initiation of supplementation using an automated chemiluminescence immunoassay (Immulite 2000, Siemens Healthcare Diagnostics). 41 dogs of 22 breeds were included with a median age of 6.2 (range 1.7-13.1) All of the dogs had symptoms of CE. Concurrent medical treatment and diet was given based on clinical judgement. Intestinal biopsies from the small and large intestine confirming chronic inflammation were available from 25/41 dogs. There was no statistically significant difference in serum cbl concentrations at baseline between the two groups (P = 0.25, Mann Whitney test). Serum cbl increased in all dogs after supplementation. In the group receiving PO supplementation, median serum cbl concentrations was 245 ng/l (150-285) at inclusion (n = 22), 970 ng/L (564-2026) after 28 days (n = 22), and 1334 ng/L (768-3921) after 3 months (n = 20). The difference between baseline and 28 days was statistically significant (P < 0.0001; Wilcoxon matched-pairs signed rank test) as well as the difference between 28 days and 3 months (<0.006; Wilcoxon matched-pairs signed rank test). In the group receiving PE cbl, mean cbl concentration was 221 AE 43 ng/L at inclusion (n = 19), 1686 AE 757 ng/L after 28 days (n = 19), and 803 AE 275 ng/L (n = 13) after 3 months. The difference between baseline and 28 days was statistically significant (P < 0.0001; paired t-test) as well as the difference between 28 days and 3 months (P = 0.0003, paired t-test). Comparing the increase in serum cbl concentrations at 28 days to baseline, the increase was significantly higher in the PE group than the PO group (mean increase 1465 AE 745 ng/L and 888 AE 449 ng/L, respectively, P = 0.004, unpaired t-test). However, after 3 months, the median (range) increase in serum cbl concentrations compared to baseline group were significantly higher in the PO than the PE group (970 ng/L (538-3663) and 608 ng/L (38-997), respectively, p = 0.0001, Mann Whitney test).

GI18

For scintigraphy, 60-second static acquisitions were performed in right and left lateral recumbency at 0, 5, 15, 30, 45, and 60 minutes following a 4 mCi Tc99M-mebrofenin solid test meal (20% of daily energy requirements), and at 30-minute intervals thereafter. Mean total counts from a gastric region of interest were measured from both images. Geometric means of the decay-corrected counts were calculated, and the amount of gastric radioactivity was expressed as a percentage of the initial count. Using nonlinear regression analysis, 25%, 50%, 75% and 90% GE times were determined. Sonographic assessment of GE was repeated three times after the same meal (non-labeled). Using the same time points, mean cross-sectional area of transverse images of the relaxed antrum was obtained and expressed as a percentage of the maximal area. The area under the curve was calculated and 25%, 50%, 75% and 90% GE times determined.
These results suggested that mosapride enhances postprandial gallbladder motility. Although the mechanisms of this effect are still to be clarified, our results suggested that mosapride enhances gallbladder motility possibly via gallbladder 5-HT 4 receptor and motilin does not play a role in this effect. Further studies are needed to support our results. Tritrichomonas foetus (Tf) is a flagellated protozoan parasite that is recognized as a significant cause of diarrhea in domestic cats with a prevalence rate as high as 30%. No drugs have been shown to consistently eliminate Tf infection in all cats. Cysteine proteases (CPs) have been identified as mediators of Tf-induced adhesion-dependent cytotoxicity to the intestinal epithelium. Thus, CPs represent novel targets for the treatment of feline trichomonosis. However, cats also produce CPs that are part of life-critical systems. Thus, parasitic CPs need to be selectively targeted to reduce the potential for host toxicity. Previous studies have demonstrated the importance of a specific CP, CP30, in mediating bovine and human trichomonad cytopathogenicity. Thus, the aims of this study were to characterize the presence of CP30 in feline Tf isolates and to evaluate the effect of targeted inhibition of CP30 on feline Tf-induced adhesion dependent cytotoxicity.
In conclusion, the results of this study provide evidence that omeprazole may be relatively safe when administered to cats for 60 days or less, but slow withdrawal of the medication is recommended based on the hypergastrinemia and rebound hyperacidity demonstrated in this pilot trial. However, further studies involving more cats are needed to confirm these results. Little is known about the behavior of food material in the esophagus during transit to the stomach in dogs with ME. The aims of our project were to standardize a contrast video fluoroscopy technique to evaluate esophageal transit time in dogs with ME while sitting in a vertical position, and to use this information to determine an appropriate duration an individual patient with ME should remain vertical after drinking and feeding.
In dogs that were accustomed to the Bailey chair, performance of contrast video fluoroscopy was technically straightforward. Significant individual variation was seen between dogs for transit time of all three ingesta types. Ingesta consistency affected transit time, but the consistency that allowed the most rapid transit time varied by individual. Mean transit time for liquid was 15.4 mins with 58% (7/12) of dogs having no clearance of liquid by 20 mins. Mean transit time for meatballs was 24.6 mins with 67% (8/12) having incomplete clearance of meatballs at 30 minutes. Mean transit time for the slurry consistency was 26.3 mins with 75% (6/ 8) of tested dogs having incomplete clearance of food at 30 mins. Three dogs were identified as having gastroesophageal reflux and one dog was found to have a concurrent swallowing disorder. Based on individual imaging results, recommendations were made to alter management strategies for 8/12 dogs; change food consistency in 50%, change time kept upright in 25% of dogs, and alter pharmacologic therapy in 17%.
Fluoroscopy performed utilizing vertical positioning in modified Bailey chairs in dogs with ME provided clear information that allowed for individual evaluation of the most ideal food consistency, anticipated behavior of ingested water, and overall esophageal transit time for ingested food material. This information permitted informed decision-making with regards to what food type appeared to be best for individual dogs, whether providing drinking water at times separate to feeding was likely to be well tolerated, and how long the dog would need to remain in a vertical position after eating or drinking. The imaging also allowed for identification of concurrent disorders affecting food tolerance and allowed for more specific management strategies in the majority of dogs. The lack of a simple and widely available test for assessing gastrointestinal motility in dogs can make diagnosing disorders difficult. Ambulatory light-based imaging is an easy to use, novel gastrointestinal imaging technique performed by oral administration of a fully automated capsule-sized camera device (ALICAM) that is propelled by natural peristalsis. The time required for passage of the capsule through the stomach and small intestine can be determined based on correlation of images with an internal clock. The aim of this study was to evaluate the feasibility of using ALI-CAM to measure gastric transit time (GTT) and small intestinal transit time (STT) in healthy dogs.
The study subjects were 10 clinically healthy dogs between 3 and 8 years of age that weighed 17-32 kg. Dogs were fasted for 16-24 hours before and 8 hours after capsule administration. Images were downloaded and independently reviewed by 3 boardcertified internists. Each reviewer determined GTT and STT. The reproducibility of the results was compared between reviewers.
Ambulatory light-based imaging can be used to assess gastrointestinal transit time in healthy dogs. Further studies are needed to determine its role in dogs with gastrointestinal disease. Ambulatory light-based imaging is a gastrointestinal imaging technique performed by oral administration of a fully automated camera device contained in a capsule (ALICAM) that is propelled by natural peristalsis. Gastric transit time (GTT) can be determined by visualization of the capsule's passage from the stomach into the duodenum and correlating images with the clock function of the device. The aim of this study was to use the ALI-CAM system to compare the GTT in healthy dogs with the GTT in sick dogs with clinical signs that could be consistent with gastric hypomotility.
The results of this study suggest that gastric hypomotility occurs in a subset of dogs presenting with retching, regurgitation, and/or vomiting. Further studies are needed to determine if these dogs would benefit from prokinetic therapy. The purpose of this study was to test the safety and feasibility of a disposable and fully automated ingestible camera system that images the gastrointestinal tract in ambulatory dogs.
Five client-owned dogs were food restricted for 24 hours before and 8 hours after capsule administration. Capsules were administered using a direct pilling technique. Normal activity and access to water were permitted throughout the study. After recovery, capsules were inspected for damage. Images were then downloaded and independently reviewed by three board-certified internists.
Capsules were successfully administered to 5/5 dogs and were recovered within 24 to 36 hours. There were no adverse events. Median (min-max) study duration was 16 (8-18) hours. Capsules recorded a median (min-max) of 19,713 (8572-51,683) images. Median (min-max) gastric and small intestinal transit times were 89 (10-110) minutes and 134 (68-177) minutes respectively. Visualization of the mucosa and image quality were described as excellent for the stomach and small bowel. The colon was poorly imaged due to retained feces.
ALI is safe and feasible in dogs. It produced high quality images throughout the majority of the gastrointestinal tract. Visualization of the colon might be feasible with additional preparation. This non-invasive technique may expand the role of imaging in dogs with gastrointestinal signs. A number of studies have demonstrated a poor correlation between histopathology and the clinical severity of chronic enteropathy in dogs, as well as the potential contribution of various immunohistochemical (IHC) markers to enhance interpretation of intestinal biopsies. Although a standardized scoring system has been developed for feline enteropathy using clinical parameters and histopathologic severity, a similar investigation, including the utility of IHC markers, has not yet been performed in cats. The purpose of this study was to identify correlations between histopathologic severity, immuno-histochemical markers, and clinical scoring of cats with chronic enteropathy.

GI27

A modified feline chronic enteropathy activity index score (FCEAI), a standard panel of gastrointestinal serum parameters, a standardized quantification of histopathology, a quantitative fecal score, and a group of immunohistochemical (IHC) markers were determined in 29 cats presented for signs consistent with chronic enteropathy. Study participants were from a client-owned population of cats experiencing diarrhea and/or vomiting of at least 3weeks duration; had no evidence of a non-gastrointestinal disease based on history, physical examination, biochemical profile, complete blood count, urinalysis, total T4, and fPLI and fTLI; and had endoscopy with biopsies and histopathology of the small intestine as part of their diagnostic work-up. Serum cobalamin and folate concentrations were also determined. Histopathologic grading and IHC quantification was performed by a single, board certified pathologist blinded to any additional information regarding the case. Immunohistochemistry was performed to identify the prevalence of cellular staining for TGF-b, CD3, Foxp3, and MAC387. The degree of cellular staining was measured at 3 random and separate areas and those 3 values were averaged to give the final immunohistochemical score for each particular marker. Biochemical parameters, FCEAI scores, and histopathologic grading scores were tested for normality using the Kolmogorov-Smirnov test, and since the vast majority of data was not normally distributed, non-parametric statistics were used for analysis.
Inclusion criteria for dogs with EPI were: a serum cTLI concentration ≤ 2.5 µg/L, age ≥ 1 year, clinical signs of EPI, and no other concurrent disease. Healthy dogs had no clinical signs of gastrointestinal disease. Fecal samples were obtained for three consecutive days, pooled, and DNA was extracted using the Power-SoilÒDNA Isolation Kit (MoBio Laboratories) for analysis of lactic acid producing bacteria by quantitative PCR. Fecal samples were deproteinized and fecal D-and L-lactate were measured by enzymatic methods (R-Biopharm, D-/L-lactic acid kit). A Kruskal-Wallis test followed by Dunn's post-test was used to compare lactate concentrations and bacterial abundances between groups. Statistical significance was set at P < 0.05. D-lactate was significantly increased in both the treated (median . Lactobacillus and Bifidobacterium were significantly increased in both treated (P = 0.0007and P = 0.0019, respectively) and untreated dogs (P = 0.0120 and P = 0.0076, respectively) with EPI when compared to healthy controls.
Inclusion criteria consisted of chronic GI clinical signs (vomiting, diarrhea, anorexia, weight loss), exclusion of non-GI diseases and discontinuation of all medications for at least 2 weeks. Biopsy specimens were obtained by gastroduodenoscopy. Helicobacter species remained undetermined in 13.3% of the patients. No HP was found in animals examined. In most dogs, the endoscopic appearance of the gastric mucosa was normal. Overall, the most common endoscopic lesions were erosions or ulcers and increased granularity of gastric mucosa. Gastritis was diagnosed histologically in 36.7% of dogs positive for Helicobacter spp. and in 41.7% of dogs without evidence of Helicobacter colonization. Severe lymphocytic-plasmacytic gastritis (11.1%) and acute purulent-necrotic gastritis (1.5%) were found in single HB infections and moderate lymphocytic-plasmacytic gastritis (14.3%) in single HH s.s. infections. An association between the presence of mixed infection (including HH s.s. and HB) and the lower severity of gastric inflammation was detected when compared to single Helicobacter species infection. This is a large study evaluating association of the individual Helicobacter species with histologically confirmed gastritis in dogs. Dogs are frequently colonized by Helicobacter spp. other than HP, and may represent a reservoir for human infection with non-HP Helicobacter spp. The combination of cytology and rapid urease test performed on gastric biopsy samples represents a reliable diagnostic method suitable for clinical practice. Competitive inhibition may occur when 2 Helicobacter species are simultaneously detected in gastric biopsies, and the presence of one Helicobacter species may interfere with the virulence of other Helicobacter species. Ambulatory light-based imaging (ALI) is a new imaging modality that allows for non-invasive endoluminal visualization of the gastrointestinal mucosa. ALI is performed by oral administration of a capsule containing a fully automated camera (ALICAM) that is propelled by natural peristalsis. The capsule is retrieved from the dogs' feces and images are downloaded. Familiarity with normal and abnormal findings is essential to accurate interpretation of these studies, but these have yet to be documented in veterinary patients. The aim of this study was to present a gallery of normal and abnormal images of the canine gastrointestinal tract using ALICAM.
ALICAM studies from 65 dogs were evaluated retrospectively by a board-certified internist. The average study consisted of 21,003 images obtained over 16 hours. Patients evaluated included 55 dogs with clinical signs of gastrointestinal disease and 10 asymptomatic dogs. The patients ranged in age from 4.5 months to 13.7 years old and in weight from 7.8 to 72 kg. Images representative of normal mucosa and common lesions were saved and logged.
Single, naturally voided fecal samples from 27 adult dogs (≥1 year of age) and 15 puppies (<1 year of age) were collected from dogs with congenital and acquired cardiac diseases. Two dogs, one puppy and one adult, had serial fecal evaluations (i.e., before and after occlusion of their patent ductus arteriosus (PDA) with an Amplatz Ò canine ductal occluder (ACDO) device). Each dog in the study had an echocardiogram performed by a board certified cardiologist or a resident under the supervision of a board certified cardiologist. Fecal samples were frozen upon collection and fa 1 -PI concentration was measured using an in-house radioimmunoassay and was compared with an established reference interval for healthy adult dogs. A fa 1 -PI concentration of ≥21.0 lg/g feces was considered abnormal.

GI34 EVALUATION OF THE EFFECTS OF PRE-CONDITIONING

Lag time (lag), time to peak (ttpeak), peak thrombin generation (peak), and endogenous thrombin potential (ETP) were measured in 41 clinically healthy dogs. Both direct jugular venipuncture and winged-needle catheter-assisted saphenous venipuncture were used to collect samples from each dog. Storage stability at À80°C was assessed over 2 months in a subset of samples. Biological variability of CAT was assessed via nested ANOVA using samples obtained weekly from 9 dogs for 4 consecutive weeks.

HM02 DEVELOPMENT AND VALIDATION OF

A novel bleeding assessment tool, DOGiBAT, was developed for canine ITP comprising nine different anatomic site-specific bleeding grades ranging from 0 (none) to 2 (severe). An online training course was developed for application of the DOGiBAT to score clinical cases. A case-based quiz set of still images was used to assess the trainees' ability to apply the DOGiBAT tool. Additionally, to assess the efficacy of the training course, 70 veterinary student volunteers were randomized to take the quiz with (n = 35) or without (n = 35) the training. All students scored all sites from all cases and site scores were considered correct if they agreed with the investigators' pre-specified classification. The frequency of correct scores was compared between trained and untrained students. A logistic regression model was used to assess the association between training and score, while adjusting for correlated responses from sites within cases.
Our study suggests that the DOGiBAT is simple scoring tool for classification of bleeding severity in dogs with ITP. A short online training course was shown to improve the accuracy and consistency of DOGiBAT scoring. Adoption of the DOGiBAT scoring system for future clinical trials of canine ITP patients will facilitate more rigorous assessment of treatment-effect based on standardized bleeding outcomes. Aspirin is commonly used in an effort to prevent thrombus formation in dogs. High-dose aspirin (10 mg/kg, PO, q 12 h) reliably inhibits platelet function and thromboxane A 2 (TXA 2 ) synthesis, but can be associated with unacceptable side effects, including inhibition of prostacyclin synthesis, which counteracts the effects of inhibition of TXA 2 . Lower doses of aspirin, in contrast, inhibit platelet function and TXA 2 synthesis, have few side effects, and allow ongoing prostacyclin synthesis. However, low-dose aspirin therapy at standard dosage (0.5-1 mg/kg, PO, q 24 h), does not reliably inhibit platelet function, a phenomenon known as "aspirin resistance". Aspirin-associated platelet dysfunction has been shown to be highly dose-dependent, suggesting that traditional "lowdoses" of aspirin may be under-dosing canine patients. Our study used incremental increases in dosages to determine the dose of aspirin that consistently inhibited platelet function and TXA 2 synthesis without inhibiting prostacyclin synthesis.
These results indicate that relative expression of BAFF mRNA is increased significantly and specifically in dogs with pITP compared to both healthy dogs and dogs with other causes of thrombocytopenia, similar to previous findings in people. Thus, BAFF may be a useful biomarker to distinguish dogs with pITP from those with other causes of thrombocytopenia. Using these primers our group has also identified a canine macrophage cell line (030-D) that expresses BAFF and is utilizing them for protein identification and quantitation as well as further characterization of BAFF mRNA splice variants. In dogs, the antiplatelet drugs aspirin (ASA) and clopidogrel may be used in an effort to prevent thromboembolism. Response to these drugs might be variable as in humans, therefore platelet function testing may be warranted to monitor treatment effect. This study aimed to evaluate the effects of ASA, clopidogrel, or combination therapy, in healthy laboratory Beagles using three platelet function tests: Multiplate Analyzer (MP), Platelet Function Analyzer-200 (PFA), and Plateletworks (PW). Secondary goals were comparison of PW using two hematology analyzers: Vetscan HM5 (using impedance) and ADVIA 2120 (using flow cytometry); and evaluation of the novel INNOVANCE P2Y cartridge for PFA to evaluate clopidogrel-induced platelet inhibition.
Six dogs were given ASA 1 mg/kg/day and placebo, clopidogrel 2 mg/kg/day and placebo, or a combination of both drugs, for 1 week each. Drugs were given in a randomized crossover design (Latin square). Each dog was randomized to one of the three treatments in three different phases such that each dog received each treatment. Investigators were blinded to the treatments. A washout period of 2 weeks occurred between phases. Blood samples were collected on days 0 and 7 of each phase, and analyzed using MP (adenosine diphosphate [ADP], arachidonic acid [AA], collagen [COL] agonists), PFA (P2Y, COL-ADP, COL-Epinephrine [EPI] cartridges), and PW (ADP, AA, COL agonist tubes) using the two hematology analyzers.

HM06

One hundred and forty-six thrombocytopenic dogs were identified, with 32 dogs not surviving to discharge. Sixty-eight dogs were excluded due to detection of an underlying infectious or neoplastic disease. Of the dogs with presumptive ITP, 11 were lost to followup and 14 dogs were still receiving prednisolone or other immunosuppressive agents. Four of these dogs remained on long-term prednisolone due to relapse when the dose was tapered.
These results failed to show an association between relapse of presumptive primary ITP in dogs and vaccination, although the possibility cannot be excluded. Future studies assessing higher numbers of dogs would be required to further assess a potential association. Aspirin, or acetylsalicylic acid (ASA), is a cyclooxygenase inhibitor that prevents the synthesis of hemostatically active prostaglandins. At low doses aspirin is used to inhibit platelet function and ultimately thrombus formation. Some human patients appear non-responsive to these antiplatelet effects, termed "aspirin resistance." This same phenomenon has been identified in dogs and could be a contributing factor to the formation of thromboemboli in some at risk patients. Accurately predicting aspirin response prior to treatment could prevent inappropriate drug therapy and devastating thrombus formation in otherwise manageable diseases.
Medical records of 32 dogs diagnosed with all forms of lymphoma, 23 dogs diagnosed with IBD and 28 control dogs, presented to the Centre Hospitalier Universitaire V et erinaire between 2006 and 2014 were retrospectively reviewed. Each blood smear was blindly re-assessed by a board-certified pathologist. A comparison between the following 3 groups: healthy dogs, dogs with IBD and dogs with lymphoma, as well as within the lymphoma group (stages 1,2,3 versus 4/5, sub-stage a versus b, and small versus intermediate/large cells) was performed for the following hematologic parameters: hematocrit, hemoglobin, erythrocyte count, mean corpuscular volume, and mean corpuscular hemoglobin concentration. The total numbers and each specific RBC anomaly were compared between these 3 groups. RBC morphologic anomalies were also categorized by the underlying pathologic process (regenerative anemia, oxidative stress, or other morphologic changes) and compared between the 3 groups.
Prevalence of anemia was significantly higher in dogs with lymphoma than dogs with IBD (P = 0.02) and healthy dogs (P = 0.0001), and significantly higher in stages 4 and 5 lymphoma as compared to stage 1, 2 or 3 (P = 0.045). There was a significantly higher total number of RBC morphological anomalies in the peripheral blood of dogs with lymphoma as compared to dogs with IBD (P = 0.02) and healthy dogs (P < 0.0001). Dogs with sub-stage b lymphoma also had a higher prevalence of total RBC morphologic anomalies as compared to sub-stage a (P = 0.04). When categorized by pathologic etiology, dogs with lymphoma had a significantly higher number of RBC anomalies associated with regenerative anemia (P < 0.0001), oxidative stress (P = 0.002), and other RBC anomalies (P < 0.0001) when compared to healthy dogs, but no significant difference when compared to dogs with IBD. The prevalence of eccentrocytes was significantly higher in dogs with lymphoma compared to IBD (P = 0.03), as well as healthy dogs (P = 0.002). No differences were noted between small, intermediate and large cell lymphoma for all parameters studied.

HM10 CHARACTERIZATION OF THE IMMUNOPHENOTYPE OF DOGS WITH IMMUNE-MEDIATED HEMOLYTIC ANEMIA.

Serum and EDTA-anticoagulated blood samples were collected from healthy control dogs (HC), dogs with primary IMHA (IMHA), and dogs with inflammatory diseases (INF). Dogs were considered to have primary IMHA if they were anemic (with hematocrit less than 35%) and fulfilled one of the following criteria: significant spherocytosis on fresh blood smear, persistent agglutination of erythrocytes after dilution in saline, or a titer of greater than 1:16 in a direct antiglobulin test. Dogs were excluded if any underlying cause of disease was detected by serum biochemical testing, imaging of the thorax and abdomen and testing for infectious agents. Dogs in the INF group had diseases that were not considered to have an immune-mediated etiology and had no previous history of autoimmune disease. Dogs in the HC group had no previous history of autoimmune disease and had unremarkable physical examinations. Owners of dogs consented for collection and use of samples, and the study was approved by an institutional ethical review committee.
Samples were obtained from 25 IMHA dogs, in addition to 20 HC and 11 INF dogs; these groups had similar age and sex distributions. There was no difference in the proportion of regulatory T cells, helper T cells or cytotoxic T cells between groups (P = 0.226), but the serum concentrations of IL-2 (P < 0.001) and TNFa (P < 0.001) were significantly greater in IMHA dogs compared to either of the other groups. The serum concentrations of IL-6 (P = 0.001) and IL-8 (P = 0.001) were significantly greater in IMHA dogs compared to HC, but there was no difference compared to the INF group. There was no difference between groups in the serum concentration of IL-10 (P = 0.731), nor in the relative expression of IL-10 (P = 0.805) or IFNc (P = 0.670). Principal component analysis revealed three major factors underlying the variability in the dataset, of which one, positively correlated with IL-2, IL-8, TNFa, blood monocyte count, serum total bilirubin concentration and negatively correlated with hematocrit, differed between IMHA and INF dogs (P = 0.001).
Immunophenotyping of dogs with IMHA revealed increased concentrations of some pro-inflammatory cytokines compared to HC and INF dogs. Characterization of factors explaining variability in all parameters suggested development of different underlying inflammatory processes in IMHA and INF groups. Further work is required to determine whether these changes are associated with survival and whether interventions that ameliorate the effects of IL-2 and TNFa might improve outcome in dogs with IMHA. Leflunomide is an immunomodulating agent labeled for the treatment of rheumatoid arthritis and psoriatic arthritis in humans. The drug inhibits the synthesis of ribonucleotide uridine monophosphate pyrimidine which ultimately lessens lymphocyte replication. This effect could be of benefit in the treatment of a variety of immune mediated diseases in dogs. The pharmacokinetics of leflunomide are known in dogs and the drug can suppress lymphocyte proliferation in this species. However, little clinical information concerning use of the drug in dogs is available. The purpose of this retrospective study was to report safety and efficacy of leflunomide for the treatment of naturally occurring immune mediated diseases in dogs.
A lower starting dose of leflunomide of 2 mg/kg/day than the current suggested dose of leflunomide of 3-4 mg/kg/day is recommended based on the findings in this study. Storage of blood products creates an unnatural environment that can accelerate product degradation and pose risk to the recipient. Eicosanoids are arachidonic acid-derived bioactive molecules that are involved in inflammation and can accumulate inside units of packed red blood cells (pRBCs). Since leukocytes and platelets are the primary sources of eicosanoid synthesis within units of pRBCs, removal of these cells via leukoreduction may decrease eicosanoid synthesis and potentially reduce the risk of transfusionrelated complications.

HM12 EFFECTS OF LEUKOREDUCTION AND STORAGE ON EICOSANOID CONCENTRATIONS IN UNITS OF CANINE

In this crossover study, units of blood were collected from 8 healthy dogs. In half of the units, leukocytes and platelets were removed via leukoreduction. Units of pRBCs were then created via centrifugation and stored for 10 or 21 days. Initial baseline plasma samples were collected after processing and before storage (Day 0). Additional samples were collected following storage for 10 or 21 days, both after removal from refrigeration and again after 5 hours at room temperature (to simulate transfusion conditions). Concentrations (ng/mL) of arachidonic acid (AA), prostaglandin E 2 (PGE 2 ), prostaglandin D 2 (PGD 2 ), prostaglandin F 2a (PGF 2a ), thromboxane B 2 (TXB 2 , stable metabolite of TXA 2 ), 6-keto-prostaglandin F 1a (6-keto-PGF 1a , stable metabolite of prostacyclin) and leukotriene B 4 (LTB 4 ) were quantified by liquid chromatography-mass spectrometry.
Prior to storage (Day 0), there were significantly greater concentrations of TXB 2 in units of leukoreduced pRBCs compared to non-leukoreduced units. In all leukoreduced samples, except for Day 10 post-transfusion, the TXB 2 concentrations decreased compared to Day 0. Except for the pre-storage samples, there was no difference in TXB 2 concentrations between leukoreduced and non-leukoreduced units. On both Days 10 and 21, there was a significant increase in 6-keto-PGF 1a concentrations for both leukoreduced and non-leukoreduced units. There was no difference in the 6-keto-PGF 1a concentrations between leukoreduced and nonleukoreduced units, except for the Day 10 post transfusion samples, in which the leukoreduced units were increased compared to the non-leukoreduced units.
Prior to storage, the PGF 2a concentrations were significantly greater in units of leukoreduced pRBCs compared to non-leukoreduced units. However, for the remaining time points, the PGF 2a concentrations decreased compared to Day 0, and there was no difference between leukoreduced and non-leukoreduced units.
In units of pRBCs, leukoreduction markedly influenced some eicosanoid concentrations. In general, removal of platelets and white cells tended to decrease production of most eicosanoids during storage. For some eicosanoids, however, this effect was counteracted by the passage of blood through the leukoreduction filter, which lead to marked increases in eicosanoid levels in units of pRBCs prior to storage. Further studies will be needed to determine what impact the complex effects of leukoreduction on eicosanoids in stored pRBCs will have on the incidence of transfusion reactions. Dog Erythrocyte Antigen (DEA) 5 is a red cell membrane receptor, and is one of the six blood groups that have been described having clinical implications 1,2 . Up until recently, blood typing was unavailable for this antigen. Previous supplies of the DEA 5 antisera had been exhausted, and the ability to create more was seriously hindered. A novel antisera was recently discovered, bringing back the ability to accurately blood type for the DEA 5 antigen.
With the re-introduction of a novel anti-DEA 5 reagent, a more complete profile of canine blood types can be achieved. This can result in better clinical decision making regarding transfusion in the canine patient, and therefore better outcomes, including the avoidance of delayed transfusion reactions when a DEA 5 incompatibility exists. Nonregenerative immune-mediated anemia (NRIMA) is a common hematologic disorder in dogs. Because the bone marrow of the affected dogs shows erythroid hyperplasia and/or erythroid maturation arrest, it has been recognized as immune-mediated destruction of erythroid precursor cells. A variety of immunosuppressive agents have been used in the treatment of this disease, but information regarding the therapeutic efficacy is limited due to the lack of prospective research. The aim of this pilot study was to assess the treatment outcome to the pre designed immunosuppressive regimen including corticosteroids, cyclosporine and mycophenolate mofetil.
8 dogs with NRIMA referred to Hokkaido university teaching hospital between 2012 and 2015 were included. Inclusion criteria in this research were a minimum 5-day history of anemia, a severe nonregenerative anemia (Hct < 20%) with absolute reticulocyte count < 60 9 10 3 /ll, and absence of any identifiable underlying disease which could induce severe anemia. The following data were collected for each dog: age, breed, sex, results of hematological analyses performed on admission, results of bone marrow examination. First treatment regimen included prednisolone (2 mg/kg/day) and cyclosporine (10 mg/kg/day) for 8 weeks. Good response were defined as an absolute reticulocyte count > 60 9 10 3 /ll or increasing Hct. Dogs that responded within 8 weeks continued to be treated with gradual tapering of dose, and re-evaluated after 6 month. Dogs that did not respond to the first regimen proceeded to second regimen including prednisolone and mycophenolate mofetil (15 mg/kg, twice a day). Similarly, good response for second regimen was determined as explained above and dogs that did not respond to both regimens were defined as poor response.
Dogs ranged 5 to 9 years old (median age, 9.5 years), and included 5 castrated males, 2 intact females and 1 spayed female. There were 6 miniature dachshunds, 1 Shih-tzu and 1 Papillon. Median Hct and absolute reticulocyte count were 16.3% and 59,640/ll, respectively. Bone marrow erythroid series were evaluated as hyperplasia in 5 dogs, euplasia in 2 dogs and hypoplasia in 1 dog. Erythroid maturation arrest was observed in all dogs. 6 of 8 dogs underwent core biopsy and 2 dogs showed myelofibrosis. 5 of 8 dogs responded to the first regimen and time to response was between 28 and 52 days. 3 dogs stopped receiving the first regimen due to anorexia that might be induced by Cyclosporine. 6 month prognoses of the other 2 dogs were complete remission (CR: Hct increased to reference range) and partial remission (PR: Hct did not reach the reference range), respectively. 3 dogs that did not respond to first regimen were proceeded to the second regimen. 1 of 3 dogs showed response after 29 days and 6 month prognosis was PR. The other 3 dogs that discontinued receiving Cyclosporine were also proceeded to the second regimen, and 6 month prognoses were CR in 2 dogs and PR in 1 dog. 2 dogs that did not show any response to both treatments had myelofibrosis.
In this research, response to both regimens could be evaluated in 6 dogs and median response time was 35 days (range 28-52). These results may be valuable for practitioners in consideration of changing immunosuppressive agent.

HM16 SERIAL EVALUATION OF THROMBOELASTOGRAPHY AND PLATELET AGGREGOMETRY IN HEALTHY DOGS.

Tissue factor activated TEG was performed in six specific pathogen free (SPF) sex and age-matched healthy Beagles at three different time points where the variables reaction time (R), clotting time (K), rate of clot formation (a), and maximum amplitude (MA) were recorded. Adenosine diphosphate (ADP)-induced and arachidonic acid (AA)-induced platelet aggregometry in addition to platelet count, hematocrit, and fibrinogen were also performed concurrently. APTT, OSPT, antithrombin activity, and D-dimer concentrations were measured on the first day of the study. A oneway random effects model was used to analyze the following variables; R, K, a, MA, area under the curve for ADP (AUC ADP ), and area under the curve for AA (AUC AA ). Variance components were created from the random effects model and used to calculate coefficients of variation for interindividual variability (CV G ) and intraindividual variability (CV I ). A repeated measures ANOVA was performed on the platelet count, hematocrit, and fibrinogen over the three time points with P < 0.05 considered significant.

HP01

Electronic medical records at Colorado State University were reviewed for cases of histopathologically confirmed gallbladder mucocele between December 2010 and January 2015. 29 cases were available and suitable for evaluation. Case data included patient signalment, clinical signs, clinicopathological abnormalities, ultrasonographic findings, histopathology, and bacterial culture of bile and liver. Formalin-fixed, paraffin-embedded gallbladder sections were mounted on charged glass slides and evaluated by FISH using a eubacterial probe (5Cy3-EUB-338) concurrently with a control probe (56 FAM-Non-EUB-338). Sections were examined on a BX51 epifluorescence microscope, and images were captured with an Olympus DP-7 camera.
Dogs undergoing cholecystectomy for treatment of gallbladder mucocele ranged in age from 6-14 years old. The most common breeds were mixed (n = 10), Shetland Sheepdog (n = 3), Miniature Schnauzer (n = 2), Pomeranian (n = 2), and Miniature Dachshund (n = 2). Ultrasound was performed in all 29 cases, with 22/29 (76%) exams concluding the appearance of the gallbladder was consistent with a mucocele. Histopathology revealed cystic mucinous hyperplasia in all cases, with associated cholecystitis noted in 13/29(45%) cases. Bacteria were detected by eubacterial FISH in 8/29 (28%) cases. Bacteria were visualized as multifocal clusters of short rods within the mucus (n = 1), adherent to the wall (n = 1), or within the parenchyma (n = 4) in 6/29 (21%) gallbladders. In 2 cases, a single bacterium was observed within the gallbladder parenchyma. Of the 6 cases with multifocal bacteria, 4 had concurrent cholecystitis. Bacterial culture of mucocele contents was positive in only 1/24 cases, yielding moderate growth of Escherichia coli. FISH analysis of this case showed diffuse colonization of mucocele contents and mucosa by E. coli.
FISH detected the presence of bacteria in 28% of mucoceles evaluated. FISH was more sensitive than bacterial culture, which was positive in only 1/8 cases with visible bacteria. Our results reveal that bacterial infection is more common than previously thought, and support the need for further studies to examine the relationship between gallbladder mucoceles, cholecystitis, and bacterial infection. After cholecystectomy a median of 27% of dogs die before hospital discharge. The cause of GBM formation is unknown. Retrospective studies identify 14-17% of dogs with GBM have a diagnosis of hypothyroidism compared to 5-9% of normal dogs. Whether hypothyroidism is an independent risk factor for GBM formation or the consequence of an underlying metabolic syndrome is unknown. This distinction has important implications regarding the cause of GBM formation and may establish a rationale for diagnosis or treatment of hypothyroidism in these dogs. We undertook a prospective case controlled study of dogs newly diagnosed with GBM formation and lacking clinical signs of hypothyroidism to test a hypothesis that occult hypothyroidism exists at a high prevalence in these dogs and that the cause may differ from classic lymphocytic thyroiditis. Dogs diagnosed with GBM by ultrasound and healthy, age, breed and sex-matched dogs with normal gallbladder ultrasound were enrolled. Exclusion criteria included clinical evidence, prior diagnosis or treatment for hypothyroidism; drugs interfering with thyroid function, intact reproductive status, or emergent disease. Testing included a CBC, chem panel, UA, thyroid function testing including serum total T4, total T3, free thyroxine by equilibrium dialysis (FT 4 ), free triiodothyronine (FT 3 ), thyrotropin (TSH) and autoantibodies to T4AA, T3AA and TGAA. Dogs were stratified by disease severity into 1) no clinical signs, 2) mild clinical disease, 3) moderate to require hospitalization, and 4) severe requiring intensive care (incl. emergent surgery). Thyroid histology of GBM dogs and controls was examined by a pathologist blinded to the diagnosis. Parametric data were analyzed by t-test and non-parametric data by Mann-Whitney U test. P < 0.05 was considered significant. Of 106 screened dogs, 38 with GBM met study inclusion criteria and ranged in age from 6-16 years (median 10.5 yrs). Breeds included 11 Shetland sheepdogs, 4 Cocker Spaniels, 3 Chihuahuas, 3 Beagles, and 16 other pure breeds. Illness severity scores were: 37% absent; 21% mild; 21% moderate; and 21% severe. Of 18 healthy dogs screened, 12 were enrolled. Dogs with GBM had significantly lower serum T 4 (median 11, range 0-44 nM), T 3 (median 0.7, range 0-1.7 nM), FT 4 (median 15, range 1-27 pM), and FT 3 (median 3, range 0-10.5 pM) compared to control dogs. Relative to control dogs, dogs with GBM frequently had severely low individual thyroid hormone concentrations. Serum T4 was below reference range (RR) in 50% of GBM dogs compared to 8% controls. Four (10.5%) GBM dogs had a T4 = 0. Serum T3 was below RR in 47% of GBM dogs compared to 25% controls. Three (8%) GBM dogs had a T3 = 0. Serum FT4 was below RR in 12% of GBM dogs compared to 0% controls. Serum FT3 was below RR in 17% of GBM dogs compared to 0% controls. Significant differences in T4AA, T3AA or mean TSH were not identified. However 23.5% of GBM dogs and only 8% control dogs had an elevated TSH. Serum TGAA was elevated in 17% of control dogs and 0% of GBM dogs. No inflammation was detected in the thyroids of any of 12 dogs with GBM undergoing autopsy. These results identify a high prevalence of occult, often severe, decreases in individual serum thyroid hormone concentrations in GBM dogs. Influence of non-thyroidal illness must be considered for some of these abnormalities. Lack of serum TGAA and absent histologic inflammation in thyroids of GBM dogs is noteworthy. These findings suggest that an alternative mechanism for abnormal thyroid function in dogs with GBM should be considered. Hepatic fibrosis is an important complication of canine chronic hepatitis. To the authors' knowledge, the interobserver agreement associated with the histological scoring of fibrosis from canine hepatic biopsy specimens has not previously been reported. Therefore, the aim of this study was to evaluate the interobserver agreement for the histological scoring of canine hepatic fibrosis.

HP04 EMBOLIZATION OF INTRATRAHEPATIC PORTOSYS-TEMIC SHUNTS IN DOGS WITH A PROTOTYPE COIL.

The prototype coil was implanted in a total of 25 dogs. Most common breeds were Hovawart (n = 8) and Golden Retriever (n = 5). Median age was 10 months (range 6 -35) and median body weight 26 kg (range 12 -48). iPSS originated from the right lateral (n = 1), right medial (n = 7) or left portal vein branch (n = 17). 19/25 dogs showed portal hypertension during shunt blocking. Unblocked and blocked shunt diameter were 5.2 -13.0 mm (median 8.3; n = 25) and 9.0 -21.0 (median 13.3; n = 24), respectively. Intervention was successful at the first (n = 24) or second (n = 1) intervention. Major complications developed in 5 dogs leading to death in 2 dogs. Minor complications were common and were mostly associated with bleeding problems. One dog was lost to follow-up after three months. Of the remaining 22 dogs, 5 experienced complications associated with change to a non-protein restricted diet: severe neurological abnormalities leading to euthanasia (n = 1), moderate symptoms leading to re-intervention (n = 2), mild symptoms which could be treated successfully by changing to a moderately protein restricted diet (n = 2). All dogs without portal hypertension showed functional closure at 3 months follow-up. In dogs with portal hypertension the cumulative proportion of functional closure reached 81% at 24 months follow-up.

HP05 INVESTIGATION OF HEPATIC COPPER ACCUMULATION IN DOGS FROM TWO TIME PERIODS

A total of 373 cases were evaluated, including 207 dogs predisposed to CCAH and 166 non-predisposed dogs. Independent of breeds groupings and inflammatory activity, mean hepatic copper concentrations in the recent time period were 225 ppm greater than those in the historical time period (P < 0.05). This effect was predominantly due an increase of 386 ppm over time in predisposed breeds (P < 0.001). The mean hepatic copper concentration of 338 ppm in non-predisposed breeds in recent years was greater than the mean hepatic copper concentration of 271 ppm in nonpredisposed breeds in the historical period, but this trend did not reach significance (P = 0.08). In the recent period, the mean hepatic copper concentration of 519 ppm in predisposed breeds with non-inflammatory liver histology was greater than the mean hepatic copper concentration of 297 ppm in non-predisposed breeds with non-inflammatory liver histology (P < 0.001). This difference was not observed in the historical time period. In both periods, dogs with hepatitis had higher hepatic copper concentrations than those without hepatitis, but this difference was magnified in recent years. In the aggregate, these findings confirm that hepatic copper concentrations have changed over time. The etiology of these changes is unknown, but suggestive of changes in environmental copper exposure. Detailed investigations of potential sources of copper exposure, namely drinking water and diet, are needed to further elucidate a potential environmental etiology. Additional investigations also are needed to determine if these changes are important for all breeds of dog. Borrelia burgdorferi (BB) and Anaplasma phagocytophilum (AP) are transmitted by Ixodes spp., and seroprevalence rates in endemic areas can be greater than 15% in dogs. A genetic predisposition to infection (Bernese mountain dogs) or clinical illness associated with BB has been suggested, particularly in people and dogs with Lyme nephritis (Retriever breeds). In vaccine studies, wild caught I. scapularis adults placed in feeding chambers and allowed to feed for seven days usually results in at least a 75% BB infection rate. Recently, an I. scapularis infestation study using a previously reported model and ticks with a similar BB carriage rate from the same collection area previously used, resulted in only a 29.2% infection rate. The purpose of this report is to describe the genetic relatedness of the dogs used in that study.

ID01 EVIDENCE FOR GENETIC PREDISPOSITON TO

A total of 10 female and 14 male purpose-bred beagles, 9-12 months of age, and negative for antibodies against A. phagocytophilum, B. burgdorferi, and E. canis (Accuplex 4 BioCD system; Antech Diagnostics), were infested for 7 days with 13 female and 12 male wild-caught I. scapularis with an estimated 48% BB infection rate and a 15% AP infection rate. Blood was collected from all dogs prior to tick attachment and then on Weeks 1-12 and Week 18 after tick attachment and screened for BB and AP antibodies as described. Dogs that seroconverted were assumed to have been infected. Statistical analyses were performed with commercial software; P ≤ 0.05 was considered statistically significant.
Antibodies against BB were detected in 7 of 24 dogs (29.2%) between 4-8 weeks post-infestation; all positive dogs were females (P = 0.0003). When positive and negative dogs were compared, there were no significant differences in the ages, number of ticks recovered, or number of fed ticks. Antibodies against AP were detected in the serum of 10 of 24 dogs (41.7%). Results using the same source of dogs, ticks, and model in previous publications were 77.8% and 55.5%% for BB and AP, respectively. The differences in infection rates between the current and historical studies were similar for AP but significantly lower in the current study for BB (P = 0.004). The most recent common ancestor (MRCA) for 6 of the 7 (86%) BB-positive dogs was either a sibling or parent. Dogs A and B were siblings and had the same father as Dog C. The mother of Dog D was the maternal grandmother of Dogs A and B. The father of Dog E was the paternal great grandfather of Dogs A, B, C. The mother of Dog F was the maternal grandmother of Dog E. The MRCA for 8 of the 17 (47.1%) BB-negative dogs was either a sibling or parent. None of the BB positive dogs had a MRCA sibling or parent in common with any of the BB negative dogs.
The POCKIT Ò iiPCR system was quick to learn, portable, and had a short run time of approximately 1 hour. There were few false-positive results, indicating that positive results are likely to represent true infections when tested in high-risk animals. Approximately 10-15% of infected dogs would be missed by the POCKIT Ò iiPCR system. However, the portability and speed with which results can be obtained may result in more infected dogs being diagnosed than in the current situation in which testing is rarely performed in dog-fighting cases due to cost and logistics of sending samples to outside laboratories. This system may also be used for B. gibsoni treatment monitoring using a hybrid approach. For example, following initial diagnosis with either the POCKIT Ò system or a commercial laboratory, the POCKIT Ò system could be used for testing during treatment. Once a negative result was obtained, a sample could be submitted to a commercial laboratory to confirm treatment efficacy. Although this study focused on mass screening for B. gibsoni, the portable iiPCR platform has potential to aid in rapid detection of a variety of infections under field conditions. The efficacy of sarolaner (Simparica TM , Zoetis) to prevent transmission of Borrelia burgdorferi and Anaplasma phagocytophilum from infected wild-caught Ixodes scapularis to dogs was evaluated in a well-controlled laboratory study. Twenty-four purpose-bred laboratory Beagle dogs seronegative for B. burgdorferi and A. phagocytophilum antibody were allocated randomly to one of three oral treatment groups: placebo administered on Days 0 and 7, sarolaner administered on Day 0 (28 days prior to tick infestation), or sarolaner administered on Day 7 (21 days prior to tick infestation). Sarolaner tablets were shaved and/or sanded based on each dog's individual bodyweight to provide a dosage of 2 mg/kg. On Day 28 each dog was infested with approximately 25 female and 25 male wild caught adult I. scapularis that were determined by random sampling to have infection rates of 57% for B. burgdorferi and 6.7% for A. phagocytophilum by PCR. In situ tick counts were conducted on Days 29 and 30. On Day 33, all ticks were counted and removed. Blood samples collected from each dog on Days 27, 49, 63, 77, 91 and 104 were tested for the presence of B. burgdorferi and A. phagocytophilum antibodies using the SNAP Ò 4Dx Ò Plus Test, and quantitatively assayed for B. burgdorferi antibodies using an ELISA test. Skin biopsies collected on Day 104 were tested for the presence of B. burgdorferi by bacterial culture and PCR. Acaricidal efficacy was calculated based on the reduction of geometric mean live tick counts in the sarolaner-treated groups compared to the placebo-treated group for each tick count.

ID04

Over a period of 3 months, veterinarians and students from RUSVM visited all 9 parishes of St. Kitts, covering a diverse geographic area with a wide diversity in ecosystem landscape. One hundred and eleven dogs were identified and visited, and were given a complete physical exam, then blood and feces were collected. Demographic data was collected, including breed, age, housing, other animals in household, evidence of ticks or fleas, deworming and vaccine administration. Blood was analyzed using a researchbased broader canine vector-borne infection serologic platform (SNAPÒ M-A, IDEXX Laboratories), which tests for specific antibodies against E. ewingii, E. canis, E. chaffeensis, Anaplasma platys, A. phagocytophilum, and Borrelia burgdorferi. This testing platform is also based on a rapid ELISA SNAPÒ technology.

ID07 DOES A DIVA TEST EXIST FOR DIFFERENTIATING FIV INFECTION FROM FIV VACCINATION?. Cynda Crawford.

Plasma samples were collected from 104 uninfected SPF cats vaccinated 3 times with a killed dual-subtype FIV vaccine (Fel-O-Vax, Boehringer Ingelheim) according to manufacturer instructions. Age at vaccination ranged from 2.5 to 13 months. The interval between vaccination and sample collection ranged from 1.75 to 14 months. The FIV-free infection status of all cats was confirmed by virus culture. Three FIV-infected cats were similarly vaccinated and tested. The plasma samples were tested in 4 commercial point-of-care assays: SNAP Ò Feline Combo FeLV Ag/FIV Ab Test, WITNESS Ò FeLV-FIV Test, Anigen Ò Rapid FIV Ab/FeLV Ag Test Kit, and VetScan Ò Feline FeLV/FIV Rapid Test. All testing was performed by personnel blinded to sample status. Each test result was independently interpreted by 2 personnel.

ID08 AN EPIDEMIOLOGICAL STUDY OF GAMMAHER-PESVIRUSES IN DOMESTIC CATS IN JAPAN. Morihiro

Tateno, Yasuyuki Endo, Masashi Takahashi. Kagoshima University, Kagoshima, Japan Gammaherpesviruses (GHVs) are members of an emerging subfamily of the Herpesvividae. The two major human GHVs, Epstein-Barr virus (EBV) and Kaposi's sarcoma-associated herpesvirus (KSHV) are clinically important pathogens because those viruses possess tumorigenesis potential especially in immune-compromised patients. In feline practice, we often experience tumor cases, especially lymphomas. Feline leukemia virus (FeLV) was a major cause for the development of lymphoma in the past days, however, the majority of lymphoma cases are shifting to FeLVfree cats. But the mechanism of tumorigenesis in these "spontaneously" developed lymphomas has not been well understood. Under this situation, GHV became a candidate which explains the lymphomagenesis in domestic cats. Recent study identified a novel GHV in domestic cats (Felis catus GHV, FcaGHV), and epidemiological surveys found that FcaGHVs are distributing in several countries including the United States, Australia, Singapore, Germany and Austria. In the present study, a molecular epidemiological study was conducted to investigate the prevalence of GHVs in Japan using large numbers of samples which were previously used to a nation-wide epidemiological survey for two feline retroviruses in Japan (Nakamura Y et al., J Vet Med Sci, 72:1051-1056, 2010).
GHV-derived DNA in blood samples was detected by degenerate nested pan-GHV PCR targeting gB gene as previously reported. Nucleotide sequences of obtained DNA fragments were determined and BLAST analysis was carried out. Phylogenic analysis was also conducted. Risk factors relating to GHV infection on clinical parameters, including age, sex, status of retrovirus infections, bite wound history and clinical signs, were statistically analyzed using univariable (Fisher's exact test) and multivariable (Logistic regression model) analyses.
GHV-derived DNA was detected in 2.6% of Japanese domestic cats. BLAST analysis revealed that all of them showed high similarity with FcaGHVs which were isolated from domestic cats in the United States. Phylogenic analysis also revealed that all Japanese isolates formed one cluster with FcaGHV. Age (> 2 years old) (Odds ratio [OR], 6.02; 95% confidential interval [CI], 0.79-46.0; P = 0.05) and feline immunodeficiency virus (FIV) infection (OR, 5.41; 95% CI, 1.93-15.1; P < 0.01) were found to be a risk factor in univariable analysis. In multivariable analysis, only FIV infection (OR, 3.74; 95% CI, 1.28-10.94; P = 0.016) was a significant risk factor.

ID09 CANINE INFLUENZA H3N2 INFECTION IN FOUR DOGS.

Anne Cohen 1 , Jill Richardson 2 , Amy Glaser 3 , Edward Dubovi 3 , Nyssa Reine-Salz 2 . 1 Chicago Veterinary Emergency and Specialty, Chicago, USA, 2 Merck Animal Health, Madison, USA, 3 Cornell University, Ithaca, USA The H3N2 influenza virus is of avian origin and was first isolated from clinically ill dogs in China in 2006 and South Korea in 2007. 1 Canine H3N2 influenza virus has been associated with severe respiratory signs and other clinical signs such as fever, reduced body weight, and interstitial pneumonia. 1 This case report presents four confirmed, naturally infected clinical cases of Canine Influenza H3N2 in the United States. The cases presented were all pet dogs that had histories of recent exposure to other dogs in a social setting including dog parks and doggie day cares. None of the dogs originated from, or had traveled to, typical regions endemic for this viral disease. All dogs were presented at different stages of illness attributed to infectious respiratory disease.
All four of the dogs were diagnosed with Canine Influenza H3N2 through PCR, three through the New York State Veterinary Diagnostic Laboratory and one through Idexx Diagnostic Laboratory. In patients with characteristic clinical features, Canine Influenza H3N2 virus infection should still be considered as differential diagnosis. A study was conducted in dogs to evaluate the efficacy of an inactivated Canine Influenza Virus (CIV) H3N2 vaccine following experimental challenge with a virulent heterologous strain of CIV H3N2, isolated from the recent CIV H3N2 outbreak in the United States. Eleven dogs, 7-8 weeks of age, were vaccinated with 2 doses of an inactivated CIV H3N2 vaccine, 3 weeks apart, and 19 dogs were vaccinated with a placebo. Two weeks after the second vaccination, all dogs were challenged intranasally with virulent CIV H3N2 and then monitored daily for 10 days for clinical signs including fever, nasal discharge, sneezing, coughing, depression, and dyspnea. Nasal swabs were collected to evaluate viral shedding, and serum samples were collected at various time points to determine antibody titers. At necropsy, lungs were scored for consolidation. Following the booster vaccination, the placebo-vaccinated control dogs remained seronegative (<10) to CIV H3N2, while 10 of the 11 vaccinated dogs developed an antibody titer to CIV H3N2 (GMT = <80; Range = <10 -320). Antibody titers in dogs from both treatment groups increased following challenge, but the increase was greater in the vaccinated dogs (GMT = >1452; Range = 40 ->10,240). Following challenge, 8 (42%) of the 19 placebo-vaccinated control dogs were euthanized prior to the 10-day post-challenge observation period due to severe clinical signs, including difficulty breathing, depression, fever, and severe coughing with retching; whereas, none (0%) of the vaccinates had to be euthanized (P = 0.014). Clinical signs were evaluated based on a weighted scoring system. The mean clinical score for the placebo-vaccinated control group was 24.9, compared to only 8.7 for the vaccinated group (P = 0.036). The placebovaccinated control group shed CIV H3N2 virus for a mean of 1.9 days, compared to 1.4 days for the vaccinated group (P = 0.507). The median lung consolidation score for the placebovaccinated control group was 7.4, compared to 0.0 for the vaccinated group (P = 0.026). Results of this study demonstrate that this inactivated CIV H3N2 vaccine significantly protects dogs against severe clinical disease and lung consolidation associated with a virulent CIV H3N2 infection. Canine infectious respiratory disease complex (CIRDC) is caused by many different viruses and bacteria. In June 2015, veterinarians around Atlanta, Georgia and also western areas of North Carolina began to notice an unusual increase in dogs presenting to their clinics with signs of infectious respiratory disease. Nasal and pharyngeal swabs from dogs exhibiting clinical signs were submitted to the Animal Health Diagnostic Center (AHDC) at Cornell University. A canine respiratory polymerase chain reaction (PCR) screening panel was utilized which allows identification of the following CIRDC pathogens: Bordetella bronchiseptica, Mycoplasma cynos, canine adenovirus 1 and 2, canine distemper, canine influenza A, H3N8 and H3N2, parainfluenza virus 5, pneumovirus and respiratory coronavirus.
Responses from 95 facilities (approximately 5% of all registered facilities in OH) of varying sizes and geographic locations were received and adequately completed to allow for analysis. Nearly all respondents (94; 99%) allowed animals in their facilities (livein, visiting, or both) , with dogs being the most frequent animal involved with facility activities (99%), followed by cats (89%), birds (74%), fish (57%), and rodents (28%). High-risk species such as reptiles/amphibians and "petting zoo farm animals (e.g., pigs, goats, chickens)" were often permitted in facilities (27% and 12%, respectively). Few facilities had restrictions targeting particular species (n = 9) or age (n = 1) of animals. Animals were reported to visit most (71%) facilities weekly or more often. One respondent (1%) reported an animal (cat) was mistreated by a resident or staff in the preceding 12 months. No known or suspected animal-associated infections were reported for the preceding 12 months. Perceived benefits of animals in facilities were high (e.g., all participants agreed/strongly agreed that "residents appeared to be happier after interacting with an animal" and "animals were useful for calming agitated residents").
The concentration of BHL, HHL and OdDHL were not affected by time (P ≥ 0.1) or AZ (P ≥ 0.2). The presence of PA in ear canal samples was affected by time (P = 0.002) but not AZ (P = 0.06). The presence of rod shaped bacteria, cocci and neutrophils in the sample were affected by time (P ≤ 0.02) but not AZ (P ≥ 0.08). Macrophages were not affected by time or AZ. Clinical signs and, therefore, the duration of the disease was influenced by time (P < 0.0001). Azithromycin had no effect on clinical signs and the duration of the disease (P = 0.5).

ID15 DETECTION OF GIARDIASIS IN DOGS:COMPARISON OF THREE RAPID DIAGNOSTIC TESTS. Hannah Bewsey, Jan

The present study compared the performance of three in-clinic tests, the SNAPÒ Giardia Test, the VetScan Ò Canine Giardia Rapid Test (Abaxis), and the Anigen Ò Rapid CPV/CCV/Giardia Antigen Test (BioNote). Canine fecal samples used were submitted to IDEXX Reference Laboratories for ova and parasite testing by centrifugal flotation. After testing, the samples were stored at À20°C. The samples were then evaluated for soluble cyst wall antigen using the Thermo ScientificTM ProSpecTTM Giardia Microplate Assay. Positive samples were defined as those testing positive by both reference methods while negative samples were defined as those testing negative on both reference methods. A total of 95 positive samples and 81 negative samples were identified and tested on all three in-clinic tests. Six samples were excluded from sensitivity/specificity analysis because they failed to produce positive control lines on the VetScan Ò Test, despite producing valid results on the other two in-clinic tests. Results were compared to the consensus reference method results for calculation of sensitivity and specificity. Using these criteria, the SNAP Giardia Test had numerically greater sensitivity and specificity than the other 2 tests. This study used another antigen test in the definition of true positive and true negative results since the samples were not fresh fecals; and not tested using a different test method like immunofluorescent antibody assay. Anaplasma phagocytophilum (Aph) and Borrelia burgdorferi (Bb) are pathogenic bacteria vectored by Ixodes spp. and are known to infect cats. While associations between Bb infections and feline clinical syndromes are unclear, studies have demonstrated Aph infection to be associated with fever, severe lethargy, and thrombocytopenia in cats. The present study aimed to evaluate the seroprevalence of antibodies to Aph and Bb in a large population of domestic cats from across the United States to better understand their risk of exposure to these agents.

ID16

A total of 5,416 samples submitted between September 2014 and February 2015 for Complete Blood Count (CBC)-inclusive test profiles were obtained from IDEXX Reference Laboratories located in Ixodes spp.-endemic areas. Samples originated from 26 states with California (24%), Illinois (19%), and Massachusetts (13%) contributing most to the study population. The population had a median age of 12.0 years (range: 1 month -25 years) with similar representation between males (51%) and females (49%). All samples were screened by Aph-specific and Bb-specific peptide immunoassays and the results were evaluated based on various test and demographic parameters.

ID17 VALIDATION OF A HIGH-THROUGHPUT SEROLOGICAL ELISA METHOD FOR FELV P27 ANTIGEN DETECTION.

Genevieve Clark, Roberta Cahill, Brendon Thatcher, Peter F. Smith, Thomas P. O'Connor, Ramaswamy Chandrashekar, Melissa J. Beall, Jesse S. Buch. IDEXX Laboratories, Inc., Westbrook, Maine, USA Feline leukemia virus (FeLV) is a globally-distributed oncogenic retrovirus that can cause fatal disease in cats. Immunoassays that detect FeLV p27 antigen and are readily used to aid in the diagnosis of FeLV have been commercially-available for more than two decades. While single-sample rapid test formats are preferred for in-clinic use, the microtiter plate format enzyme-linked immunosorbent assay (ELISA) is the methodology most commonly employed in reference laboratory settings to facilitate batch testing. However, current commercially-available microtiter plate format ELISAs for FeLV are not well suited for this purpose because they rely on manual wash steps, drop-wise addition of reagents from reagent bottles, and visual interpretation of results. The purpose of this study was to validate a microtiter plate format ELISA designed specifically to maintain performance while enhancing throughput in a reference laboratory setting.

ID18

A total of six young laboratory-reared beagles that were infected by AP, but no other vector borne agents, as evidenced by detection of AP DNA in blood by PCR assay (FastPanel, Antech Diagnostics) and AP antibodies in serum (Accuplex 4 BioCD system; Antech Diagnostics) after infestation with wild-caught I. scapularis were selected for study. On Week 20 after I. scapularis infestation, the dogs were placed into two Groups, each containing one AP PCR positive, AP antibody positive dog and two AP PCR negative, AP antibody positive dogs. Group 1 was administered doxycycline at 5 mg/kg, PO, twice daily for 4 weeks. After sample collection the morning of Week 24, all dogs were administered prednisolone at 2 mg/kg, PO, daily for 2 weeks. The dogs were evaluated daily for clinical abnormalities and by CBC, AP PCR and AP antibody assay Weeks 24-28. Sera from 5 dogs was assayed for AP antibodies on Week 48.
The 2 dogs with anemia, thrombocytosis and neutropenia on the day prednisolone was initiated were clinically normal, AP PCR negative, and were normal in all other CBC evaluations, suggesting these results were spurious. The one dog in Group 2 that was positive for AP DNA on Week 25, was also positive on Week 24 prior to prednisolone administration. None of the other samples assessed by CBC and AP PCR assay after initiation of prednisolone administration showed evidence for activation of AP infection, suggesting that this protocol is unlikely to adversely affect dogs with chronic AP infection in the field. Whether other immune suppressive protocols would activate chronic AP infection remains to be proven. The only dogs that became AP seronegative by Week 48 were administered doxycycline, suggesting treatment may aid in the elimination of chronic AP infection, even after prednisolone administration. The objective of this study was to determine the proportion of feline calicivirus (FCV) field isolates that would be neutralized in vitro by sera from cats that were routinely vaccinated with a product containing two caliciviruses. Serum from cats hyperinoculated against 2 feline caliciviruses (FCV) has been shown to cross neutralize a greater proportion (90.2%) of 61 FCV field isolates from the United States when compared to serum from cats hyperinoculated with 1 FCV isolate (23.0%) At 8 and 11 weeks of age, each of the 44 kittens was administered an experimental vaccine containing modified-live panleukopenia, modified-live feline herpesvirus 1, and non-adjuvanted inactivated calicivirus strains FCV-255 and FCV-DD1. Serum was collected from each kitten 7 and 14 days after the second dose of vaccine and pooled by day.
All fecal samples that were submitted to a commercial laboratory (ANTECHÒ Diagnostics) between 2010 and 2015 for evaluation with a PCR panel of assays that amply DNA of a select group of infectious agents including Giardia spp. (dogs and cats), Cryptosporidium spp. (dogs and cats), and Cryptosporidium felis (cats only) were analyzed. Descriptive, univariable, and multivariable logistic regression analyses were conducted to assess associations amongst age, sex, region, and season and the probability of testing positive to either Giardia spp. or Cryptosporidium spp. in pet dogs and cats.
Cryptosporidium spp. DNA was amplified from feces of 336 of 1,762 Giardia spp. DNA positive dogs (19.1%) and 132 of 843 Giardia spp. DNA positive cats (15.7%) . Of the 843 Giardia spp. DNA positive cats, 97 (11.8%) were positive for Cryptosporidium felis DNA. Logistic regression models results showed that age (young) and region were associated with presence of Giardia spp. DNA and Cryptosporidium spp. DNA in feces of dogs and cats. In addition, presence of Giardia spp. DNA in feces of dogs varied by season.
The results indicate that Giardia spp. DNA, Cryptosporidium spp. DNA, and Cryptosporidium felis DNA are commonly amplified from feces of dogs and cats, coinfections are common, and positive rates can vary by age, season, and region. Either of the agents should be suspected more highly in young animals. Additional studies will be required to evaluate for associations of positive test results with clinical findings and to determine the likelihood dogs or cats are carrying zoonotic Giardia spp. or Cryptosporidium spp. Tick-borne diseases, including Lyme disease, ehrlichiosis, and anaplasmosis, are becoming increasingly prevalent as tick distributions expand through climate change, wildlife migration, and increased relocation of companion animals. Granulocytic anaplasmosis is caused by the obligate intracellular bacterium Anaplasma phagocytophilum, which is transmitted by Ixodes spp. of ticks. Both dogs and humans are susceptible to this infection. In dogs common signs of the infection, such as lethargy, anorexia, lameness and fever, are non-specific. Differential diagnosis is supported by evidence of morulae or DNA, especially in acute cases, and by detection of specific antibodies using IFA or ELISA-based tests. 1 A related but distinct bacterium, A. platys, also infects dogs and causes infectious cyclic thrombocytopenia. 2 The present study compared the performance of two in-clinic rapid tests, the SNAP Ò 4Dx Plus Ò Test (IDEXX), and the VetScan Ò Canine Anaplasma Test (Abaxis). Random canine samples sourced from endemic regions for A. phagocytophilum were characterized by A. phagocytophilum Immunoflourescence assay (IFA) which was performed by a commercial reference laboratory. A. platys-specific samples were sourced from dogs living in the southwest, an A. platys-endemic region. These samples were characterized using A. platys-specific peptide ELISA. 3 The characterized samples were then blinded and randomized before testing with the two in-clinic rapid tests. Results are shown in the table below.
Notably, the VetScan Ò Test failed to detect more than 60% of the medium to high titer (1:400 and above) A. phagocytophilum samples. Client-owned cats in Moscow, Russia can be exposed to fleas and ticks and some develop fever, hemolytic anemia, thrombocytopenia, gingivitis, and other clinical problems noted in cats of other countries. However, limited data are available concerning prevalence rates for infectious agents common to eastern and western Europe. The overall objective of this study was to determine the estimated prevalence of select feline infectious disease agents in a convenience sample population of cats from Moscow, Russia.
None of the cats were positive for DNA of Anaplasma spp. or Ehrlichia spp. but many of the cats showed evidence of exposure or current infection with one of more of the other agents. Bartonella spp. antibodies were most common in serum and Candidatus M. hemominutum (Mhm) DNA was most common in blood.
Toxoplasma gondii IgG and Bartonella spp. IgG titers ranged from 1:64 to 1:2048. Of the 81 cat samples evaluated in all assays, single, dual, triple, and quadruplicate infections were documented in 31, 11, 2, and 2 samples, respectively. Using this subset of samples, 57.8% of the cats had evidence of infection or exposure to one or more of the select agents.
The results from this sample population suggest cats in Moscow are exposed to intermediate hosts or undercooked meat (T. gondii), fleas (Bartonella spp. and hemoplasmas), and come in direct contact with infected cats (hemoplasmas, FeLV and FIV). These infectious agents should be on appropriate clinical differential lists, processed foods should be fed, ectoparasite control should be maintained, and feline-feline contact should be avoided when possible. Coxiella burnetii is a rickettsial pathogen with serious zoonotic implications (Q fever) transmitted by direct contact and vectored by numerous tick species. Cats infected C. burnetii have been implicated in human infections in Australia and the United States but minimal information exists concerning routes of feline infection. Recently, C. burnetii DNA was isolated from Ctenocephalides felis collected from wildlife in Cyprus; this flea commonly infests cats. The purpose of this study was to evaluate select groups of fleas from cats in the Australia and United States for the presence of C. burnetii DNA using a previously optimized PCR assay.

Species

Coxiella burnetii DNA was not recovered from fleas collected from cats in eastern Australia or the southern United States. The results suggest that fleas infesting domestic cats in these regions are not important vectors for C. burnetii and are unlikely to play a role in the transmission of the organism to humans with Q fever. Canine infection with Dirofilaria immitis, the parasitic agent responsible for causing heartworm disease, is easily and safely prevented via the periodic administration of one of a number of preventative agents. Many veterinarians in Oregon do not routinely recommend the use of heartworm prophylactics; therefore, dogs are commonly not on a preventative. Confusion regarding the use of heartworm preventatives in Oregon is understandable, given the paucity of incidence data in our region. The decision between a veterinarian and a client on whether or not to start a preventative is often made based on a cost-benefit analysis, where only the cost is known. The purpose of this study was to evaluate the risk of developing heartworm infection amongst unprotected dogs in Northwestern Oregon. This information will enable veterinarians and their clients to make more informed decisions regarding heartworm prevention.

ID25

Between March 2010 and August 2015 owners of all dogs surrendered to the Oregon Humane Society filled out a short questionnaire relating to their dogs' histories. This questionnaire was used to determine the eligibility of each dog for this study. Criteria for inclusion in the study included that the dogs: (1) had been in the possession of the surrendering party since they were 6 months of age or younger; (2) were at least 12 months of age; (3) had no history of travel outside of the state of Oregon; and (4) were never administered heartworm preventative. Dogs that did not meet all four of these criteria were excluded from the study. Additional information gathered from the records included breed, age, sex and spay/neuter status, location (by zip code), and the average number of hours the dog spent outdoors. If a dog did meet the study criteria then blood was drawn and tested for the presence of D. immitis using a commercially available ELISA test (IDEXX Snap 4Dx Plus).
The findings of this study suggest that the likelihood of unprotected Northwestern Oregon dogs developing heartworm infection is currently very low. Heartworm transmission prediction models based on climate data (Knight/Lok) have shown that the climate conditions in the area can support heartworm transmission. Data compiled by the Companion Animal Parasite Council (Capcvet.org) confirm that Dirofilaria immitis has been diagnosed within the state of Oregon, although only 0.18% of all positive cases of heartworm in the U.S. are in the state of Oregon. It is important to consider that these data reflect a point in time. As the population of Oregon continues to grow, more heartworm infected dogs will invariably arrive. Changes in climate patterns will also influence the transmission of this disease. Continued monitoring and vigilance are recommended. In the last several years, clinical illness in a number of people in Baja California Sur, M exico, has been suspected to be associated with rickettsial disease agents such as Rickettsia rickettsii, Rickettsia felis, and/or Ehrlichia species. The objective of this study is to determine the prevalence of select vectors and vector-borne disease agents carried by companion dogs of the region.

ID26 ECTOPARASITES AND VECTOR-BORNE PATHOGENS OF

Vector-borne agents detected to date likely reflect common exposure to R. sanguineus, as this tick vectors each of the PCRconfirmed agents. Further information will be gained by completion of the PCR assay analysis of the blood, fleas, and ticks. Canine group settings, locations or events where dogs temporarily come together in a shared environment (e.g., shows, sporting events, dog parks) pose an increased risk for infectious disease transmission. Despite this increased risk, few guidelines exist to provide recommendations for reducing disease risk in these settings. During 2014-2015 a panel of canine infectious disease experts reviewed the current literature and drafted a set of 44 evidence-based recommendations for prevention of infectious diseases for dogs in group settings. In August 2015 a survey of attendees at the AKC Canine Health Foundation Parent Organization conference was completed to determine agreement with and perceived barriers to these recommendations. The 15-minute self-administered survey was provided to 238 Conference attendees and consisted of a series of Likert-type and open-ended questions (online and paper format) seeking feedback on 22 of the recommendations. The survey was completed by 185 individuals (78%), and all responses were reviewed, summarized, and open-ended comments categorized by theme. Respondents self-identified as: participants, judges, and breeders in a variety of local, national, and international canine group events. Most respondents (> 40%) agreed with all but three of the panel's recommendations, yet a majority of respondents stated the recommendations would be difficult or very difficult to implement in their setting (primarily dog shows). Common survey result themes related to difficulty of implementation included: administrative concerns (cost, human resources/manpower), enforcement issues, ethical concerns, privacy concerns, and strong need for official outreach to promote awareness and education related to canine infectious diseases. Survey responses identified needs for: further refinement of recommendations to aid comprehension and clarity (especially around ecto-and endoparasite control), and education to promote culture changes related to disease risk prevention. In order to raise awareness of canine infectious disease in group settings amoungst event participants, attendees, and organizers; an online freely available canine infectious disease risk calculator tool is being developed. Mycoplasma species are one of the most common infectious causes of conjunctivitis in cats. Mycoplasma felis is commonly implicated as a primary pathogen, but other Mycoplasma species have also been detected in clinically ill cats. Findings from previous studies using conventional PCR (cPCR) to investigate the role of Mycoplasma species in causation of feline conjunctivitis have been mixed as Mycoplasma can be carried by apparently normal cats. Therefore, the purpose of this study was to determine if increasing severity of conjunctivitis in cats correlates with higher Mycoplasma species copy numbers using qPCR.

NM01

Specific antibodies against canine parvovirus type 2 (CPV2) and E. coli were obtained in eggs from hens vaccinated separately against one of the mentioned agents 3 . Egg yolk was flash-dried and tested for the presence of CPV2 and E. coli-specific antibodies. Hyper-immune solution was then prepared by mixing egg powder with a commercial milk replacer (Babydog Milk, Royal Canin, Aimargues, France; 1 g of egg powder with CPV2 antibodies and 1 g of egg powder with E. coli antibodies with 12 mL of reconstituted milk). A total of 334 puppies from 16 different breeds, enrolled in one breeding kennel, were included in the study. Depending on the expected adult body weight, puppies were classified into small breed dogs (S; adult weight < 25 kg), and large breed dogs (L; > 25 kg). Within each litter and taking into account the birth weight, puppies were assigned into supplemented or control group. Each puppy from the supplemented group received orally 1.5 mL/100 g bw of hyper-immune solution at once within the first 8 hours after birth. Puppies from the control group received at the same dose (1.5 mL/100 g) and time after birth (< 8 h) the milk replacer only. All puppies were weighed at birth and at 7, 14, and 21 days of life. Linear mixed models (MIXED procedure; SAS Institute Inc., Cary, NC, USA) with litter modeled as a random effect were performed to determine the variables affecting birth weight and weight gain during the neonatal period: breed size (small; large), age (0-7; 7-14; 14-21 days), supplementation (supplemented or control group). All the interactions between mentioned fixed effects were also tested.
In our study large breed puppies supplemented at birth with the hyper-immune egg yolk had greater weight gain during the entire neonatal period. Retarded growth at the early stage of life increases the risk of morbidity and mortality in puppies 4 . Thus it could be hypothesized that better growth in supplemented puppies reflects a better health. Nevertheless, further studies are needed in order to confirm our findings in other breeding kennels and on large number of individuals.
1 Poffenbarger et al., 1991; 2 "Canine health product containing antibodies against canine parvovirus type 2" WO2015004181 A1. Decline in cerebral glucose metabolism is a common feature of aging in people and animals including rats, dogs, and monkeys, and it is closely associated with age-dependent cognitive decline. We had confirmed in a previous study that dietary medium chain triglycerides (MCTs) can enhance cognitive functions in dogs by providing the brain with an alternative energy source. In this study, we investigated the effects of dietary MCTs on voluntary activity in dogs and cats.
In the cat study, sixteen middle-aged and senior cats were fed 100% of their maintenance energy requirements (MERs) with a control diet for one week, and then were switched to the MCTcontaining diet for 28 days. Their daily activity was monitored during the feeding trial with activity monitors. In the dog study, twenty senior dogs were fed 100% of their MERs with a control diet for 6 days, and then were assigned to one of 2 MCT-containing diets with 10 dogs per diet for 27 days. Then the dogs were switched to the alternative MCT diet for 32 days. Daily activity was recorded with activity monitors during the feeding trial. In cats, the MCT-containing diet significantly increased both daytime ( Excessive weight gain is associated with metabolic and hormonal changes that predispose cats to insulin resistance and other disorders. The adipokine leptin is an important regulator of energy metabolism, and circulating leptin concentrations correlate with fat mass in cats and other species. In cats, leptin is associated with insulin resistance, independent of bodyweight. Although dietary factors such as energy density, fat content and feeding pattern increase the risk of obesity, there is limited information on how dietary factors such as feeding, fasting, and macronutrient content influence leptin concentrations in cats. The aims of this study were to compare leptin concentrations in lean, healthy cats fed diets high in protein, fat and carbohydrate, to assess associations between leptin concentrations and insulin, glucose, and NEFA concentrations, and to determine the effects of feeding pattern on baseline and postprandial leptin concentrations.
Distributions of leptin variables in the ad libitum and mealfeeding tests were similar across dietary groups after consumption of the washout diet for 5 weeks. Mean baseline concentrations (average of À30 and À5 minutes values), mean concentrations over 24 hours (mean 24-h), and peak leptin concentrations in the meal feeding test varied significantly by diet (overall P < 0.001 to 0.027). Baseline, mean 24-h and peak leptin concentrations for the high fat diet were significantly higher than for the high protein diet, and for baseline and mean 24-h, for the high carbohydrate diet. A similar pattern was observed in the ad libitum feeding test with highest leptin concentrations in cats consuming the high fat diet. In general, leptin concentrations after consuming the high carbohydrate diet were not significantly different from the high protein diet, but if they differed, leptin concentration for the high carbohydrate diet was higher compared to the high protein.

NM06

WHEN FED FOODS WITH SIMILAR PALATABILITY, CATS CHOOSE 30%, DOGS 23% OF CALORIES AS PROTEIN. Dennis Jewell 1 , Jodi Vondran 1 , Melissa Vanchina 1 , Jean Hall 2 . 1 Hill's Pet Nutrition, Inc., Topeka, KS, USA, 2 Oregon State University, Corvallis, OR, USA Dogs and cats have specific taste preferences that are influenced by macronutrient composition and physical characteristics of food, as well as presence or absence of specific taste imparting compounds (palatability enhancers). This study investigated the relationship of food choice on macronutrient composition and that choice on subsequent plasma amino acid concentrations. Experimental foods for dogs and cats were individually prepared to have similar palatability through manipulations of factors independent of macronutrients. Foods with similar palatability preference for their respective populations as a whole were then investigated for intake choice using 20 dogs with varying body fat composition (mean, 3.9 kg; SD, 0.9 kg; range, 2.4-6.2 kg;) and 27 cats (mean body fat composition, 1.4 kg; SD, 0.9 kg; range 0.3-2.2 kg). Subsequent effects of food choice on serum amino acids were also determined. Four completely balanced foods were available to dogs and cats at all times. The four varied in macronutrient content: Food 1: high protein (33% of calories from protein for dogs; 42% for cats); Food 2: high fat (54% of calories from fat for dogs; 44% for cats); Food 3: high carbohydrates (56% of calories from carbohydrate for dogs; 52% for cats); and Food 4: blended macronutrients. Dogs on average chose to consume 23.0% calories from protein (SD, 1.5%; range 20-26%) whereas cats chose 30.3% of calories from protein (SD, 4%; range 24-38%). Body composition influenced their choice of calories from protein. In dogs, there was a positive relationship between body fat and percent calories consumed as protein (r = 0.56; P = 0.02). There was no relationship between lean body mass and percent calories consumed as protein (r = 0.06; P = 0.82). Overall, dogs with high body fat showed the highest preference for dietary calories from protein. In cats, both body fat and lean body mass were negatively associated with calories consumed as protein (P = 0.04; P = 0.02, respectively), with a positive association for the interaction (P = 0.02). Overall, cats with high body fat and high lean body mass showed the highest preference for dietary calories from protein. Dogs had higher circulating amino acid concentrations of threonine, methionine, isoleucine, and valine (P < 0.05) compared with cats, the later which had higher circulating amino acid concentrations of isoleucine, valine, tyrosine, phenylalanine, and histidine (P < 0.05). In both dogs and cats there was no effect of protein intake on circulating essential amino acid concentrations. Increased intake of calories as protein was associated with increasing plasma ornithine (dogs only) and citrulline (dogs and cats) concentrations (P < 0.05). In summary, given the opportunity to choose between foods with similar palatability, cats chose to consume 30.3% and dogs 23.0% of their calories as protein. Although there were species differences between circulating amino acid concentrations indicating metabolic shifts associated with protein intake, there were no changes in circulating essential amino acid concentrations. The purpose of this double masked, controlled multicenter clinical trial was to determine the impact of a food with ingredients designed to have skin and coat health benefits and immune modulating effects on clinical signs of seasonal atopic dermatitis versus a control food. Dogs with a history of seasonal dermatitis, but without current clinical signs consistent with atopy (CADESI and pruritis scores = 0) were recruited in the spring from 11 general practices in the United States. Dogs currently on foods designed for adverse food reactions were excluded. Eligible dogs were randomly assigned to Test or Control groups and evaluated by their veterinarian at 0, 4, 8, 12 and 16 weeks. Consistent with accepted standards of care, treatment was not withheld, therefore, prescribed medications and dosing information was collected; only oclacitinib which was not uniformly available to all practices was disallowed. Veterinarians graded skin lesions using a modified CADESI (0:absent -4:severe for erythema, alopecia, excoriations and lichenification for 27 body sites, max score of 432) and a pruritis score (0:absent -4:severe).
This study is the first to describe the recurrence rate of seasonal atopy in dogs from general practices as part of a grade 1, multicenter clinical trial. Onset of clinical signs resulted in medical intervention in the majority of dogs. Statistical power in this study was likely influenced by the generally low CADESI and pruritis scores combined with medical intervention. In addition, the interval to medical intervention was likely influenced by the study schedule which promoted frequent evaluations and case management. This data suggests that dogs with seasonal atopy my benefit from frequent evaluations. This data also suggests altered management of dogs fed the Test food as medication use was lower and the interval prior to the onset of systemic antihistamines and/or glucocorticoids was longer. A food designed for skin and coat health benefits and immune modulating effects can be part of a proactive atopy management program and may result in less medication needed. Retrospective feline studies have shown SDMA is more sensitive than sCr for early CKD detection. Our objective was to compare SDMA and sCr for detection of early CKD in a prospective, longitudinal study of older dogs.
SDMA and traditional serum and urine clinicopathologic tests were measured biannually for four years in 43 dogs with an initial median age of 8.9 years. Persistent increases in UPC, sCr, or SDMA or decreases in USG triggered renal ultrasound (US) and GFR (plasma iohexol clearance) evaluations. Necropsies were performed whenever possible when dogs died or were euthanized.

NU02 SDMA CORRELATES BETTER WITH CREATININE THAN HIGH THROUGHPUT IMMUNOTURBIDOMETRIC CYS-TATIN C ASSAY IN FELINE SERUM. Polina Prusevich, Julie

Thirty-four dogs with CKD and 10 healthy dogs were included. Dogs with CKD were staged according to International Renal Interest Society (IRIS) guidelines. A human FGF-23 ELISA was validated for canine samples, showing linearity and intra-and interassay coefficients of variation <7%. Values are presented as median (range). Plasma FGF-23 concentrations in healthy dogs and dogs with IRIS stages 3 and 4 CKD were 315 pg/mL (211-449), 2,302 (455-24,409) and 7,733 (2,520-41,265), respectively (P < 0.01).
These findings suggest that FGF-23 concentrations rise in canine CKD before other traditional markers of RHPT do. Consequently, FGF-23 concentrations should be considered as an early indicator of disease progression, RHPT, and potential target for therapeutic intervention. Erythropoiesis-stimulating agents are currently recommended for veterinary patients with anemia secondary to chronic kidney disease (CKD). Darbepoetin alfa (darbepoetin) has replaced epoetin due to a three-fold longer half-life allowing for less frequent dosing. Also, darbepoetin is perceived to be less likely to elicit formation of anti-erythropoietin antibodies resulting in pure red cell aplasia (PRCA). A previous study examined the efficacy of darbepoetin in cats with CKD; 14 of 25 cats responded, with adverse events observed in 12 cats, including two with possible PRCA. This retrospective study evaluated response to darbepoetin therapy, protocols for administration, and potential adverse events in dogs with CKD.
Thirty-four dogs with IRIS stage 3 or 4 CKD met the inclusion criteria and their medical records were reviewed. Data recorded included CBC, serum biochemistry panel, reticulocyte count, serum iron parameters, systolic blood pressure, packed red blood cell (pRBC) transfusion, iron supplementation, darbepoetin dose and frequency of administration, comorbidities, adverse events, and survival.
Darbepoetin is an effective treatment for anemia secondary to CKD. A dosing interval of >21 days is not effective at maintaining a response to therapy. Further studies are needed to define the most effective dose. PRCA was a possible adverse event in 1/34 dogs (2.9%). Vitamin D-binding protein (VDBP) is the main carrier of vitamin D metabolites into circulation, and it is freely filtrated through glomerulus, and the complex VDBP-25-OH-vitamin D is uptaken by megalin-cubilin receptors in proximal tubules. Once the presence of VDBP in urine, it may indicate proximal tubular dysfunction/injury as well as interstitial tubular fibrosis. Also it still remains to be proven whether loss of VDBP in urine may lead to vitamin D deficiency and its contribution to the progression of chronic kidney disease (CKD) in dogs. The hypothesis and the aim of this study were to investigate whether VDBP could be previously detected in urine of dogs with CKD regardless of urinaryto-protein ratio (UPC) values, and it will be an early and specific marker of proximal tubular injury. Forty dogs according to IRIS staging of CKD (stage 1 n = 10; stage 2 n = 10; stage 3 n = 10; stage 4 n = 10) were enrolled, various breeds and age, and with no proteinuric-associated disease. Nine clinically healthy dogs, different breeds and age, composed the control group. Dogs with CKD were classified according to IRIS guidelines based on UPC as nonproteinuric (UPC<0.2), borderline proteinuric (UPC= 0.2 to 0.5) and proteinuric (UPC>0.5). Western blotting was performed to investigate VDBP in canine urine (Anti-Vitamin D Binding Protein antibody, ab95469, Abcam© 1:500). No urinary VDBP was detected in control dogs and UPC was 0.14 AE 0.04 (meanAESEM) and min= 0.028 and max= 0.41. In CKD dogs, urinary VDBP was observed since the early stages of CKD, stages 1 and 2, and the UPC was 0.39 AE 0.29; 0.02-3.01 and 0.33 AE 0.12; 0.14-1.14 (mean AESEM; min -max), respectively. In stage 1 CKD dogs, urinary VDBP was detected in 7 out of 10 dogs and in 4 of those 7 dogs, UPC was into the non-proteinuric range, 2 dogs in borderline range and only one was proteinuric. In stage 2 CKD dogs, VDBP in urine was noticed in 9 out of 10 CKD dogs and 5 of them had UPC < 0.2, one dog was in borderline (UPC= 0.42) and 3 dogs were slightly proteinuric (UPC of 0.67, 0.72 and 1.14). In CKD dogs in stage 3, UPC was 1.51 AE 0.54 (0.07-4.57) and VDBP in urine was immunodetected in 8 out of 10 dogs and in 3 of those 8 dogs, UPC was ≤ 0.28 and the left 5 dogs were proteinuric. All CKD dogs in stage 4 were proteinuric (UPC = 4.37 AE 0.47; 1.4-6.94) and showed VDBP in urine. In conclusion, the immunodetection of VDBP in urine noticed in the early stages of CKD in dogs, mainly in stages 1 or 2, associated with UPC into non or borderline proteinuric range, it reinforces that urinary VDBP could be a potential early marker of kidney injury as well as to detect the specific segment (proximal) of the nephron affected. Progressive interstitial nephritis (IN) is the primary cause of feline chronic kidney disease which is considered to be the leading cause of death in adult cats. The Crandell Rees feline kidney (CRFK) cell line is commonly used to grow feline herpesvirus 1 (FHV-1), calicivirus, and panleukopenia virus used in (FVRCP) vaccine production. Previous studies have shown that cats administered FVRCP vaccines parenterally develop antibodies against CRFK lysates and alpha enolase (glycolytic pathway enzyme in all mammalian cells) and these antibodies can bind to feline renal cell lysates. In addition, three of six cats that were hyperinoculated (11 injections) with CRFK lysates over two years had IN on renal biopsy collected two weeks after the last booster. The primary objective of this study was to determine whether IN could be induced over 16 weeks by repeatedly administering a market leading parenteral FVRCP vaccine to potentially use as a short term model to study biomarkers associated with IN in cats.

NU05

A total of six (three male; three female) one-year-old purpose bred cats were included in the IACUC approved study. The cats were previously maintained as unvaccinated controls in a FHV-1 study at five months of age and none of the cats had clinical signs of FHV-1 when enrolled in the current study. On Week 0, blood, serum, urine were collected for biomarker assays and a wedge kidney biopsy for histopathological evaluation and alpha-enolase immunohistochemistry was obtained. After sample collection on Week 0 and again on Weeks 2, 4, 6, 8, 10, 12, and 14, all cats were administered a commercially available FVRCP vaccine shown previously to induce anti-CRFK and anti-enolase antibodies and samples were collected for biomarker assays. After sample collection on Week 16, similarly sized renal biopsies were made, avoiding the previous biopsy sites. Haematoxylin and eosin stained sections were provided to two board-certified pathologists that were masked to the timing of the biopsies. Anti-CRFK and antienolase antibodies levels in serum were determined by ELISAs.

NU07 COMPARISON OF VISUAL AND AUTOMATED INTERPRE-TATION OF URINARY DIPSTICKS WITH GLUCOSE:CREA-TININE RATIO AND GLUCOSE CONCENTRATION.

Urine samples submitted to the Auburn University Clinical Pathology Laboratory for urinalysis and that were naturally glucosuric (n = 39) were included. Multistix (Bayer) were used according to manufacturer instructions and were read visually by a single trained investigator ("visual reading") and by the CLINITEK 50 analyzer (Bayer; "automated reading"). Urine glucose and urine creatinine concentrations were measured using a Hitachi 911 Chemistry Analyzer (Boehringer Mannheim Corp.); the glucose measurements were considered to be the true concentration (gold standard). Spearman's rank order correlation was used to evaluate the relationship between: 1. glucose concentration and automated readings; 2. glucose concentration and visual reading; 3. automated and visual readings; 4. automated readings and UGCR; and 5. visual readings and UGCR. Significance was set at P < 0.05. Correlation was classified as excellent for r = 0.93-0.99, good for r = 0.8-0.92, fair for r = 0.59-0.79 and poor for r < 0.59. Reference intervals that correlated with the colors on the test strip were devised based on the package insert (trace=76-175 mg/dL; 1 + = 176-375 mg/dL; 2 + = 376-750 mg/dL; 3 + = 751-1500 mg/dL; 4 + = >1500 mg/dL).
Our results indicate that despite a good correlation between automated and visual reading of Bayer Multistix, automated reading may be better. Interestingly the automated readings tend to underestimate glucose concentration while with visual reading, overestimations are more common. The UGCR is a valid way to assess urinary glucose excretion; however, it offers no advantage over measurement of glucose concentration in urine. Acute pancreatitis (AP) and acute kidney injury (AKI) are common co-morbidities in both dogs and humans. In human patients the presence of AP with AKI is a negative prognostic factor; however, a similar relationship has not been evaluated in dogs. When considering hemodialysis for management of AKI in dogs, this prognostic information could be important for therapeutic decisions. Canine pancreas-specific lipase (SPEC) is used commonly to support the diagnosis of AP in dogs. The purpose of this study was to investigate elevated SPEC on the outcome of dogs with AKI treated with hemodialysis.
Medical records were evaluated retrospectively from August 2011 to June 2015 to identify dogs presented to the UC Davis VMTH that received intermittent hemodialysis for management of AKI and also had a SPEC (Spec cPL TM , IDEXX Laboratories) measured during the course of therapy. Outcome was assessed at 30 days from admission, and dogs were classified as died/euthanized or alive. Surviving dogs were stratified further into non dialysis-dependent or dialysis-dependent. An elevated SPEC was defined as ≥ 400 mcg/L. Median values of SPEC were compared with a Mann-Whitney U test for each outcome category, above. Categorical data were compared with a Fisher's exact test. A Pvalue of <0.05 was considered significant. Data are presented as median [range] .
Forty-three client-owned dogs were included. Median initial serum creatinine (iSCr) was 8.5 [3.6 -16.6] mg/dl and median age at presentation was 5.6 [0.70 -13.5] years. Eighteen were male neutered, 14 were female spayed, 4 were intact male, and 7 were intact female dogs. At 30 days, 30/43 dogs (70%) were alive (iSCr, ] mg/dl) and 13/43 (30%) had died or been euthanized (iSCr, ] mg/dl). Nine of 30 dogs (30%) were still dialysis dependent at 30 days (iSCr, ] mg/dl), and 21/30 (70%) were non dialysis-dependent (iSCr, -1001] mcg/L, P = 0.008). The proportion of dogs alive at 30 days with SPEC ≥ 400 mcg/L during the treatment course was not different from alive dogs with SPEC < 400 mcg/L (71% versus 68%, P = 1.0). In addition, the proportion of the non dialysis-dependent dogs with SPEC ≥ 400 mcg/L during the treatment course was not different from non dialysis-dependent dogs with SPEC < 400 mcg/ L (42% versus 58%, P = 0.36). These results suggest the presence of pancreatitis assessed as SPEC ≥ 400 mcg/L did not affect survival outcome at 30 days. However, pancreatitis may be a more common co-morbidity in dogs with more severe (dialysis-dependent) AKI and it may influence renal recovery. Bacterial urinary tract infection (UTI) is common in dogs, however the prevalence of subclinical bacteriuria (SBU) in healthy dogs has recently been demonstrated to be substantially lower. Following standards created by human infectious disease specialists, there has been more initiative put towards identifying and characterizing SBU in dogs. Although it has been considered a risk factor for UTI, the prevalence of UTI and SBU in dogs with chronic kidney disease (CKD) is not known. The objectives of this study were to retrospectively evaluate the prevalence of positive urine cultures in dogs with CKD, and to further categorize these patients as having SBU, clinical UTI, or pyelonephritis.
Medical records were reviewed retrospectively from 1/2010-7/ 2015 for dogs with a diagnosis of CKD who had a urine culture submitted. The diagnosis of CKD was based on International Renal Interest Society (IRIS) guidelines. Patients were excluded if they had another reason to develop UTI, namely endocrine disease (hyperadrenocorticism, diabetes mellitus), urolithiasis, urinary incontinence, urinary bladder neoplasia, had a nidus for infection such as ureteral stent or subcutaneous ureteral bypass, or were receiving systemic immunosuppression. Based on previously reported definitions, patients were placed into one of the following three categories based on the review of their medial record: SBU, pyelonephritis, or UTI. The frequency of negative culture, SBU, pyelonephritis, and UTI were compared across IRIS stages using Fisher's exact analysis. P < 0.05 was considered significant for all comparisons.
Two hundred and eighty-two cultures were submitted on 195 patients during the study time period. Forty-seven cultures performed on 13 patients were excluded due to the presence of ureteral stents (5 patients), chemotherapy (2 patients), diabetes mellitus (2 patients), transitional cell carcinoma (2 patients), urinary incontinence and cystic calculi (1 patient each). A total of 235 cultures submitted on 182 patients were included in the final analysis. There were 8 intact males, 52 castrated males, 5 intact females, and 117 spayed females. There were 40 positive urine cultures (17.0% of all cultures) obtained on 33 patients (18.1% of all dogs). Of all positive cultures, the most frequently determined diagnosis was SBU (18 cultures, 45%), followed by pyelonephritis (16 cultures, 40%), then UTI (6 cultures, 15%). There were no statistically significant observed relationships between any IRIS stage and diagnosis (P = 0.635). The number of positive cultures, regardless of patient diagnosis, was not statistically significant between IRIS stages (P = 0.432). Escherichia coli was the most frequently observed isolate (29/40 cultures, 72.5%).
In this population of dogs with CKD the prevalence of dogs with positive urine culture was 18.1%. The most frequently observed diagnosis associated with a positive culture was SBU. Increasing IRIS stage of CKD was not found to be associated with a higher frequency of any particular diagnosis. E. coli was the most commonly observed isolate. Despite the rigorous criteria for defining SBU, pyelonephritis, and UTI in the current study, it is possible that some patients were incorrectly categorized. Prospective research is needed to determine if antibiotic therapy is indicated in dogs with CKD and SBU. Additionally, renal biomarkers may allow for better characterization of patients with SBU and occult pyelonephritis and improve antibiotic stewardship. The aim of this study was to report combined experience of two institutions placing subcutaneous ureteral bypass systems (SUBs). We compared our data to historical data from cats with ureteral stents for ureteral obstruction. Hypotheses included 1) Patients with SUBs would have the same or better 7-14 day and 3-4 month post-operative creatinine as historic ureteral stent patients, 2) SUB patients would experience the same or improved peri-operative, post-operative and recurrent urinary tract infection rates as ureteral stent cats, 3) clinical signs related to implants would be similar or better with the SUB system versus ureteral stenting.
Adult cats were enrolled retrospectively, subsequent to the 2012 alteration to SUB placement procedure. All cats were managed per clinician preference pre-operatively. Flushes of SUB systems were performed every 3-6 months for the majority of patients.
Records from 19 cats from Purdue and Munich Universities were reviewed. Median age was 9 years at Purdue and 7.7 years at Munich. There were 12 spayed female and 7 castrated male cats; 13 domestic short-haired cats, 2 domestic longhaired cats, and three other breeds. 17% of cats had preexisting IRIS Stage II CKD. Lethargy, vomiting and inappetence were the most common clinical signs (>25% of patients). Median renal pelvic diameter in affected kidneys was >0.5 cm at both institutions. Median creatinine immediately post-operatively was 2.6 mg/dL (0.9-2.3 mg/ dL) at Purdue and 1.43 mg/dL at Munich. Major peri-operative complications were uncommon; one cat had cardiac arrest and another was transfusion dependent. Post-operative complications included 2 cats with fluid overload and pleural effusion. 8/19 cats required a blood transfusion in the peri-or post-operative period. The median creatinine was 3.6 mg/dL at 3 months and 1.8 mg/dL at 6 months. 3 patients had repeat surgeries; median time to repeat surgery was 225 days. Causes of repeated surgery were SUB port obstruction or encrustation of SUB tubing. Total mortality was 21% with a median disease free interval of 180 days and a 1 year survival of 83%.
In all, 31/500 (6.2%) samples were positive. Most infections were a single organism (n = 27); four contained multiple organisms. E. coli was the most common species (58% of isolates) followed by Enterococcus species (25% of isolates). Positive specimens were more likely to be from female cats (n = 24) versus male (n = 7) (P = 0.0054). Patient age and body weight were not associated with increased risk of occult UTI. Median urine specific gravity was similar for the positive group (1.022) and controls (1.024). The prevalence of proteinuria on dipstick testing was also similar (84% v 80%). Occult UTIs were strongly associated with bacteriuria (60% v 6%; odds ratio 24.3 [CI: 9.0-65.7]; P < 0.0001) or pyuria (67 v 19%; odds ratio 8.7 [CI: 3.7-20.5]; P < 0.0001) but not microscopic hematuria. Positive specimens were significantly more likely to have an abnormal sediment exam (odds ratio 13.5 [CI 3.9-45.5]; P < 0.0001).
Dogs with TCC/PCA had significantly higher urinary S100A8/ A9 and S100A12 concentrations than all other groups of dogs (all P < 0.016) except for dogs with UTI, whereas uCalR was significantly higher in TCC/PCA dogs compared to only those with a UTI (P < 0.001). Treatment-na€ ıve TCC/PCA patients and UTI dogs were best distinguished by a uCalR of ≥9.1 (sensitivity: 91%, specificity: 60%). A urine canine S100A8/A9 concentration (normalized against the urine specific gravity [USG]) of ≥109.9 had the highest sensitivity (96%) and specificity (66%) to diagnose treatment-na€ ıve TCC/PCA dogs from all other groups of dogs, and the specificity increased to 75% if a UTI had been excluded. Using a urine S100A8/A9 concentration (normalized against USG) of ≥109.9 to screen dogs ≥6 years of age for TCC/PCA (estimated prevalence: <1%) yielded a PPV of 4% and a NPV of 100%.

NU13 SURVEY OF SUBCUTANEOUS FLUID PRACTICES IN CATS WITH CHRONIC KIDNEY DISEASE. Crystal Cooley 1 , Sarah

An anonymous web-based survey was advertised via list serves, websites and social media. Owners of 468 cats with CKD participated. 87% of cats were 10 years or older. Cats were IRIS stage I (1%), II (20%), III (37%), IV (17%), and unknown (25%).

NU15 EVALUATION OF THE EFFECT OF URINE DIP VERSUS URINE DRIP ON MULTITEST STRIP RESULTS. Alisdair

Boag, Craig Breheny, Adam Gow. Royal (Dick) School of Veterinary Studies, Edinburgh, UK Standard operating procedures (SOPs) have been established for in-clinic laboratory tests, such as the World Health Organisation guidelines for packed cell volume. No independent evidence based guidelines exist for dipstick urinalysis, however manufacturer's instructions state to dip the stick into urine. In veterinary medicine, occasionally urine samples of such small volume are obtained that this is impractical and dripping urine on the stick from a pipette or syringe is performed. This also has the advantage of conserving the sample for additional tests.

NU16 POSITIVE IMPACT OF NUTRITIONAL INTERVENTIONS IN CLIENT-OWNED DOGS WITH IRIS STAGE-1 CHRONIC KIDNEY DISEASE.

Jean Hall 1 , Dale Fritsch 2 , Maha Yerramilli 3 , Edward Obare 3 , Murthy Yerramilli 3 , Dennis Jewell 2 . 1 Oregon State University, Corvallis, OR, USA, 2 Hill's Pet Nutrition, Inc., Topeka, KS, USA, 3 IDEXX Laboratories, Inc., Westbrook, ME, USA A prospective 12-month clinical trial was performed in clientowned dogs with IRIS stage 1 chronic kidney disease (CKD) to measure their ease of transition to a commercial renal protective food and to assess the effects of food on renal biomarkers and quality of life attributes. Dogs with IRIS stage 1 CKD (n = 36) were transitioned over 1 week from various grocery brand foods to a modified protein, low phosphorus, antioxidant enriched test food (Prescription DietÒ k/dÒ, Hill's Pet Nutrition, Inc.). At months 0, 3, 6, 9, and 12 owners completed a questionnaire to assess their pet's acceptance of the test food and their dog's quality of life. Renal biomarkers, including serum creatinine (Cr), blood urea nitrogen (BUN), and symmetric dimethylarginine (SDMA), and urinalysis parameters, including urine specific gravity (USG) and urine protein:creatinine ratio (UPC), were measured. Of the 36 dogs initially enrolled, 35 (97%) transitioned to the test food. Pets moderately or extremely liked the test food 88% of the time, ate most or all of the food 84% of the time, and were moderately or extremely enthusiastic while eating 76% of the time. Dogs consuming test food showed a decrease in serum Cr and BUN concentrations across time (both P = 0.01) and a decrease in serum SDMA concentration and USG across time (both P = 0.09). All serum renal biomarkers (Cr, BUN, SDMA) were decreased (P ≤ 0.05) from baseline at 3 months, and remained decreased from baseline at 12 months in dogs completing the study (n = 20). This study shows that dogs with IRIS stage 1 CKD readily transition to a commercial renal food, and decreasing serum biomarker concentrations suggest improvement in kidney function. In addition, owners reported improvement in a consortium of overall health and quality of life attributes (P < 0.01) and hair and coat quality attributes (P < 0.01). Decreased glomerular filtration rate in chronic kidney disease (CKD) reduces renal phosphorus excretion, leading to increased total body phosphorus retention and eventually hyperphosphatemia. In addition of promoting parathyroid hormone synthesis, hyperphosphatemia stimulates the synthesis of fibroblast growth factor 23 (FGF-23), a phosphaturic/hypophosphatemic hormone. There are no reports on serum FGF-23 in healthy dogs or in dogs with CKD. The goal of this study was to measure serum FGF-23 concentrations in healthy dogs and in dogs with CKD, and to determine its association with serum phosphorus concentrations and severity of renal disease based on IRIS staging of CKD. We hypothesized that serum FGF-23 will be increased in dogs with CKD, that its concentrations will be proportional to CKD severity, and that it will be an early marker of altered phosphorus metabolism.
A total of 17 dogs of various breeds and age with CKD (n = 6 in IRIS stage 2; n = 11 in IRIS stage 3) that were being fed a renal diet, and 15 healthy dogs of different breeds and age, being fed a maintenance diet, were included. Dogs with CKD were followed for up to 12 months or until death. A human-specific FGF-23 ELISA was validated for this study, showing linearity and 2.8% and 8.8% intra and inter-assay coefficients of variation, respectively. Serum FGF-23 concentrations were higher in CKD dogs in stages 2 and 3 on presentation (2,605.4 AE 872.0 pg/mL; meanAESEM) compared to