Objective—To evaluate the effects of a dexmedetomidine constant rate infusion (CRI) and atropine on changes in global perfusion variables induced by hemorrhage and volume replacement (VR) in isoflurane-anesthetized dogs. Animals—8 adult dogs. Procedures—Each dog was anesthetized twice, with a 2-week interval between anesthetic sessions. Anesthesia was maintained with 1.3 times the minimum alveolar concentration of isoflurane with and without dexmedetomidine (1.6 μg/kg, IV bolus, followed by 2 μg/kg/h, CRI). Dogs were mechanically ventilated and received an atracurium neuromuscular blockade during both sessions. During anesthesia with isoflurane and dexmedetomidine, atropine was administered 30 minutes before baseline measurements were obtained. After baseline data were recorded, 30% of the total blood volume was progressively withdrawn and VR was achieved with an equal proportion of autologous blood. Results—Following hemorrhage, cardiac index, oxygen delivery index, and mixed-venous oxygen saturation were significantly decreased and the oxygen extraction ratio was significantly increased from baseline. The anaerobic threshold was not achieved during either anesthetic session. When dogs were anesthetized with isoflurane and dexmedetomidine, they had a significantly lower heart rate, cardiac index, and mixed-venous oxygen saturation during VR than they did when anesthetized with isoflurane alone. Plasma lactate concentration, mixed venous-to-arterial carbon dioxide difference, base excess, and anion gap were unaltered by hemorrhage and VR and did not differ between anesthetic sessions. Conclusions and Clinical Relevance—Results indicated that the use of a dexmedetomidine CRI combined with atropine in isoflurane-anesthetized dogs that underwent volume-controlled hemorrhage followed by VR did not compromise global perfusion sufficiently to result in anaerobic metabolism.

Objective-To characterize the electrocardiographic features of the atrial repolarization (Ta) wave in dogs with third-degree atrioventricular (AV) block. Sample-ECGs of 36 dogs with third-degree AV block and no identifiable structural heart diseases. Procedures-Standard 12-lead ECGs were acquired with a digital system, and measurements were manually edited. Results-A Ta wave was detectable in all dogs for at least 1 ECG lead. The Ta wave had negative polarity in leads I, II, III, and aVF and positive polarity in leads aVL and aVR, with a mean electrical axis of -114.26°. Mean duration and mean amplitude of the Ta wave in lead II were 140.2 milliseconds and -0.09 mV, respectively, with the ratio for the Ta-to-P wave duration of 2.3 and the ratio of Ta-to-P wave amplitude of -0.35. Significant correlations were found between the Ta wave duration and duration of the P-Ta interval, Ta wave amplitude and the ECG lead, Ta wave duration and body weight, and duration of the P-Ta interval and atrial rate. Measurements of the Ta wave were repeatable. Conclusions and Clinical Relevance-Measurements of the Ta wave in dogs with third-degree AV block were repeatable. The values for the Ta wave reported here can be used as reference values for dogs with AV conduction disturbances and an echocardiographically normal atrial size. Further studies are needed to validate these results in dogs with structural heart diseases.

Objective-To determine whether increasing the viscosity of a standard hemoglobin-based oxygen-carrying solution (HBOC) would offset its associated vasoconstrictive effects and result in improved microvascular perfusion in healthy splenectomized dogs with experimentally induced hemorrhagic shock. Animals-12 male American Foxhounds. Procedures-Each dog underwent anesthesia and splenectomy. Shock was induced by controlled hemorrhage until a mean arterial blood pressure of 40 mm Hg was achieved and maintained for 60 minutes. Dogs were then randomly assigned to receive either a standard or hyperviscous HBOC (6 dogs/group). Sidestream dark-field microscopy was used to assess the effects of shock and HBOC administration on the microcirculation of the buccal mucosa and the jejunal serosa. Video recordings of the microcirculation were collected before shock was induced (baseline) and at intervals up to 180 minutes following HBOC administration. Vascular analysis software was used to compute microcirculatory variables. Results-Compared with baseline findings, hemorrhagic shock resulted in decreases in all microvascular variables in the buccal mucosa and the jejunal serosa. At all time points following HBOC administration, microvascular variables were similar to initial values and no significant differences between treatment groups were detected. At all time points following HBOC administration, blood and plasma viscosities in dogs treated with the hyperviscous solution were significantly higher than values in dogs receiving the standard solution. Conclusions and Clinical Relevance-In splenectomized dogs with experimentally induced hemorrhagic shock, administration of a hyperviscous HBOC did not significantly affect microvascular variables, compared with effects of a standard HBOC. Microcirculatory flow returned to baseline values in both treatment groups, suggesting that marked HBOC-associated vasoconstriction did not occur.

Objective—To investigate clinical use of proton magnetic resonance spectroscopy (1H MRS) and to compare metabolic brain bioprofiles of dogs with and without hepatic encephalopathy. Animals—6 dogs with hepatic encephalopathy and 12 control dogs. Procedures—Conventional MRI and single-voxel 1H MRS were performed with a 3-T magnet. Images for routine MRI planes and sequences were obtained. Single-voxel 1H MRS was performed with a point-resolved sequence with a short echo time (35 milliseconds) and voxel of interest placement at the level of the basal ganglia. Metabolites of interest included the glutamine-glutamate complex (sum quantification of glutamate and glutamine), myoinositol, N-acetyl aspartate, total choline, and creatine. Data were analyzed with postprocessing fitting algorithm software, and metabolite concentration relative to water and ratios with creatine as the reference metabolite were calculated. Results—Compared with control dogs, dogs with hepatic encephalopathy had specific changes, which included significantly higher concentration relative to water of the glutamine-glutamate complex and significantly lower concentration of myoinositol. Choline and N-acetyl aspartate concentrations were also slightly lower in dogs with hepatic encephalopathy than in control dogs. No differences in creatine concentration were detected between groups. Conclusions and Clinical Relevance—1H MRS aided in the diagnosis of hepatic encephalopathy in dogs, and findings supported the assumption that ammonia is a neurotoxin that manifests via glutamine-glutamate complex derangements. Use of 1H MRS may provide clinically relevant information in patients with subclinical hepatic encephalopathy, equivocal results of bile acids tests, and equivocal ammonia concentrations or may be helpful in monitoring efficacy of medical management.

The objective of this study was to validate non-equilibrium gravitational field-flow fractionation (GrFFF), an immunotag-less method of sorting mesenchymal stem cells (MSCs) into subpopulations, for use with MSCs derived from equine muscle tissue, periosteal tissue, bone marrow, and adipose tissue. Cells were collected from 6 young, adult horses, postmortem. Cells were isolated from left semitendinosus muscle tissue, periosteal tissue from the distomedial aspect of the right tibia, bone marrow aspirates from the fourth and fifth sternebrae, and left supragluteal subcutaneous adipose tissue. Aliquots of 800 × 10(3) MSCs from each tissue source were separated and injected into a ribbon-like capillary device by continuous flow (GrFFF proprietary system). Cells were sorted into 6 fractions and absorbencies [optical density (OD)] were read. Six fractions from each of the 6 aliquots were then combined to provide pooled fractions that had adequate cell numbers to seed at equal concentrations into assays. Equine muscle tissue-derived, periosteal tissue-derived, bone marrow-derived, and adipose tissue-derived mesenchymal stem cells were consistently sorted into 6 fractions that remained viable for use in further assays. Fraction 1 had more cuboidal morphology in culture when compared to the other fractions. Statistical analysis of the fraction absorbencies (OD) revealed a P-value of < 0.05 when fractions 2 and 3 were compared to fractions 1, 4, 5, and 6. It was concluded that non-equilibrium GrFFF is a valid method for sorting equine muscle tissue-derived, periosteal tissue-derived, bone marrow-derived, and adipose tissue-derived mesenchymal stem cells into subpopulations that remain viable, thus securing its potential for use in equine stem cell applications and veterinary medicine.

Objective-To assess effects of body position on direct measurements of intra-abdominal pressure (IAP) and abdominal perfusion pressure (APP) in horses anesthetized with total intravenous anesthesia (TIVA). Animals-9 healthy adult horses. Procedures-Instrumentation in unsedated standing horses involved insertion of an arterial catheter for blood pressure measurements and 3 intraperitoneal cannulas (left flank, right flank, and ventral abdomen) for IAP measurements. Baseline values were measured for heart rate, respiratory rate, systolic arterial blood pressure, mean arterial blood pressure (MAP), diastolic arterial blood pressure, and IAP. Horses were medicated with xylazine, and pressures were measured again. Anesthesia was induced with ketamine-diazepam and maintained with a ketamine-guaifenesin infusion. Horses were positioned twice into left lateral recumbency, right lateral recumbency, or dorsal recumbency. Hemodynamic pressures and accessible abdominal pressures were measured for each recumbency position. The APP was calculated as MAP - IAP. Differences in IAP, MAP, APP and sedation (standing horses) or body position (anesthetized horses) were compared by means of repeated-measures ANOVA or paired t tests. Results-Baseline hemodynamic and IAPs were not different after xylazine administration. Ventral abdomen IAP and MAP were lower for horses in dorsal recumbency than in right or left lateral recumbency. Ventral abdomen APP remained unchanged. For lateral recumbencies, flank IAP was lower and APP was higher than pressure measurements at the same sites during dorsal recumbency. Conclusions and Clinical Relevance-Body position affected IAP and APP in healthy anesthetized horses. These effects should be considered when developing IAP acquisition methods for use in horses with abdominal disease.

Objective-To evaluate tissue oxygen saturation (Sto2) by use of near-infrared spectroscopy in experimental acute hemorrhagic shock and resuscitation in dogs. Animals-14 healthy adult purpose-bred Beagles. Procedures-Dogs were anesthetized with isoflurane via facemask, anesthesia was maintained with propofol and rocuronium bromide, and dogs were mechanically ventilated to maintain normocapnia. Dogs were studied under normovolemia (baseline), hypovolemia with target mean arterial blood pressure < 40 mm Hg achieved and maintained steady for 10 minutes (hypovolemia T1), then 20 minutes later (hypovolemia T2), following resuscitation with shed blood (after transfusion), and after administration of 20 mL of hetastarch/kg (hypervolemia). Conditions were executed sequentially during a single anesthetic episode, allowing stabilization between states (10 minutes). Hemoglobin concentration, mean arterial blood pressure, arterial blood gas concentrations, cardiac index, oxygen delivery indexed to body surface area, and Sto2 were monitored. Results-From baseline to hypovolemia T1, there was a significant reduction in mean +/- SD oxygen delivery index (619 ± 257 mL/min/m2 to 205 ± 76 mL/min/m2) and StO2 (94 ± 4.4% to 78 ±12.2%). Following resuscitation, Sto2 (80 ± 8.5% vs 92 ± 6.45%) and oxygen delivery index (211 ± 73 mL/min/m2 vs 717 ± 221 mL/min/m2) significantly increased, returning to baseline values. Hypervolemia had no effect on Sto2 or oxygen delivery index. A strong correlation (r = 0.97) was detected between mean oxygen delivery index and Sto2 across all time points. Conclusions and Clinical Relevance-Under the conditions of this study, there was a strong correlation between Sto2 and oxygen delivery, suggesting that Sto2 may be used to estimate oxygen delivery.

Objective—To assess the microcirculatory effects of IV fluid administration in healthy anesthetized dogs undergoing elective ovariohysterectomy. Animals—49 client-owned dogs. Procedures—Dogs were sedated, and anesthesia was induced with propofol and diazepam and maintained with isoflurane in oxygen. Dogs received lactated Ringer's solution (LRS) IV at rates of 0, 10, or 20 mL/kg/h. Videomicroscopy was used to assess and record effects of LRS administration on microcirculation in the buccal mucosa. Measurements of microcirculatory (total vessel density, proportion of perfused vessels, microcirculatory flow index, and perfused vessel density by vessel size [< 20 μm, ≥ 20 μm, and all diameters]) and other physiologic variables (heart rate, Doppler-measured blood pressure, oxygen saturation as measured by pulse oximetry, capillary refill time, and body temperature) were compared among groups at baseline (immediately after anesthetic induction), 30 and 60 minutes afterward, and overall. Results—Neither the proportion of perfused vessels nor microcirculatory flow index varied among treatment groups at any time point, regardless of vessel size. For vessels < 20 μm in diameter and for all vessels combined, total and perfused vessel density were similar among groups. For vessels ≥ 20 μm in diameter, total vessel density was significantly greater in the 20 mL/kg/h group than in other groups, and perfused vessel density was significantly greater in the 20 mL/kg/h group than in the 0 mL/kg/h group, when all time points were considered. Other physiologic variables were similar among groups. Conclusions and Clinical Relevance—Total and perfused vessel density of vessels ≥ 20 μm in diameter (mostly venules) were greatest in dogs that received 20 mL of LRS/kg/h. Further research is required to evaluate clinical importance of these findings.

Objective—To evaluate the postoperative analgesic effects of epidural administration of morphine and neostigmine, either alone or in combination, in dogs. Animals—30 dogs undergoing orthopedic surgery on a pelvic limb. Procedures—Anesthetic protocols were standardized. At the end of surgery, 10 dogs each received 1 of 3 epidural treatments: morphine (0.1 mg/kg), neostigmine (5 μg/kg), or morphine plus neostigmine (0.1 mg/kg and 5 μg/kg, respectively). Postoperative pain scores and the need for rescue analgesia were evaluated for 24 hours. Results—Pain scores were higher in the neostigmine group, compared with scores for the morphine-neostigmine group, at 2 and 24 hours after surgery and higher in the morphine group than in the morphine-neostigmine group at 2 and 4 hours. During 24 hours, rescue analgesia was provided for 4, 7, and 2 of 10 dogs each in the morphine, neostigmine, and morphine-neostigmine groups, respectively. The number of dogs given rescue analgesia was significantly different among groups at 2, 3, 4, and 6 hours after surgery. Dogs in the morphine and morphine-neostigmine groups had a lower probability of receiving rescue analgesia within 24 hours than did dogs in the neostigmine group. Conclusions and Clinical Relevance—When administered epidurally, morphine alone or in combination with neostigmine provided effective postoperative analgesia in most dogs after orthopedic surgery, whereas neostigmine alone did not. Findings for this study suggested a potential role for neostigmine as an adjuvant for epidural analgesia in dogs undergoing orthopedic surgeries on the pelvic limbs.

Objective-To determine the usefulness of retina samples for detection of disease-associated prion protein by use of a commercially available enzyme immunoassay (EIA) intended for rapid identification of sheep and cattle with transmissible spongiform encephalopathies (TSEs). Samples-Retina, brainstem at the level of the obex, and retropharyngeal lymph node samples obtained from 15 TSE-inoculated sheep (scrapie [n = 13] or transmissible mink encephalopathy passaged through a bovid [2]); retina and brainstem samples obtained from 11 TSE-inoculated cattle (transmissible mink encephalopathy passaged through a bovid [7] or classical BSE [4]); and negative control tissue samples obtained from 2 sheep and 2 cattle that were not inoculated with TSEs. Procedures-Tissue samples were homogenized and analyzed for detection of abnormally folded disease-associated prion protein with a commercially available EIA and 2 confirmatory assays (western blot analysis or immunohistochemical analysis). Results-Retina sample EIA results were in agreement with results of brainstem sample EIA or confirmatory assay results for negative control animals and TSE-inoculated animals with clinical signs of disease. However, TSE-inoculated animals with positive confirmatory assay results that did not have clinical signs of disease had negative retina sample EIA results. Retina sample EIA results were in agreement with brainstem sample immunohistochemical results for 4 TSE-inoculated sheep with negative retropharyngeal lymph node EIA results. Conclusions and Clinical Relevance-Results of this study suggested that retina samples may be useful for rapid EIA screening of animals with neurologic signs to detect TSEs.