Objective-To characterize and quantitatively assess the typical pulmonary anatomy of healthy adult alpacas with multidetector row CT. Animals-10 clinically normal adult female alpacas. Procedures-CT examination of the thorax was performed before and after IV administration of iodinated contrast medium in sedated alpacas in sternal recumbency. Measurements of the trachea, bronchi and related blood vessels, and selected vertebrae as well as the extent and density of lung parenchyma were performed with a Digital Imaging and Communications in Medicine (DICOM) viewer. Morphometric and quantitative data were summarized. Results-Separation of individual lung lobes could not be identified, except for the accessory lung lobe. In all alpacas, both lungs extended farther caudally at the medial aspect than at the lateral aspect. The right lung extended farther in both cranial and caudal directions than did the left lung. The branching pattern of the bronchial tree varied only slightly among alpacas and consisted of 1 cranial bronchus and 3 caudal bronchi bilaterally, with a right accessory bronchus. Luminal diameters of first-generation bronchi ranged from 3 to 9 mm. Mean +/- SD parenchymal lung density was −869 +/- 40 Hounsfield units (HU) before contrast injection and −825 +/- 51 HU after contrast injection. Mean difference in diameter between bronchi and associated arteries or veins was 0.8 +/- 0.9 mm. Conclusions and Clinical Relevance-Knowledge of the typical anatomy of the lungs and bronchial tree in healthy alpacas as determined via CT will aid veterinarians in clinical assessment and bronchoscopic evaluation of alpacas.

Objective—To determine relative concentrations of selected major brain tissue metabolites and their ratios and lobar variations by use of 3-T proton (hydrogen 1 [1H]) magnetic resonance spectroscopy (MRS) of the brain of healthy dogs. Animals—10 healthy Beagles. Procedures—3-T 1H MRS at echo times of 144 and 35 milliseconds was performed on 5 transverse slices and 1 sagittal slice of representative brain lobe regions. Intravoxel parenchyma was classified as white matter, gray matter, or mixed (gray and white) and analyzed for relative concentrations (in arbitrary units) of N-acetylaspartate (NAA), choline, and creatine (ie, height at position of peak on MRS graph) as well as their ratios (NAA-to-choline, NAA-to-creatine, and choline-to-creatine ratios). Peak heights for metabolites were compared between echo times. Peak heights for metabolites and their ratios were correlated and evaluated among matter types. Yield was calculated as interpretable voxels divided by available lobar voxels. Results—Reference ranges of the metabolite concentration ratios were determined at an echo time of 35 milliseconds (NAA-to-choline ratio, 1.055 to 2.224; NAA-to-creatine ratio, 1.103 to 2.161; choline-to-creatine ratio, 0.759 to 1.332) and 144 milliseconds (NAA-to-choline ratio, 0.687 to 1.788; NAA-to-creatine ratio, 0.984 to 2.044; choline-to-creatine ratio, 0.828 to 1.853). Metabolite concentration ratios were greater in white matter than in gray matter. Voxel yields ranged from 43% for the temporal lobe to 100% for the thalamus. Conclusions and Clinical Relevance—Metabolite concentrations and concentration ratios determined with 3-T 1H MRS were not identical to those in humans and were determined for clinical and research investigations of canine brain disease.

Objective—To investigate the in vitro differentiation of canine bone marrow stromal cells (BMSCs) into functional, mature neurons. Sample—Bone marrow from 6 adult dogs. Procedures—BMSCs were isolated from bone marrow and chemically induced to develop into neurons. The morphology of the BMSCs during neuronal induction was monitored, and immunocytochemical analyses for neuron markers were performed after the induction. Real-time PCR methods were used to evaluate the mRNA expression levels of markers for neural stem or progenitor cells, neurons, and ion channels, and western blotting was used to assess the expression of neuronal proteins before and after neuronal induction. The electrophysiological properties of the neuron-like cells induced from canine BMSCs were evaluated with fluorescent dye to monitor Ca2+ influx. Results—Canine BMSCs developed a neuron-like morphology after neuronal induction. Immunocytochemical analysis revealed that these neuron-like cells were positive for neuron markers. After induction, the cells’ mRNA expression levels of almost all neuron and ion channel markers increased, and the protein expression levels of nestin and neurofilament-L increased significantly. However, the neuron-like cells derived from canine BMSCs did not have the Ca2+ influx characteristic of spiking neurons. Conclusions and Clinical Relevance—Although canine BMSCs had neuron-like morphological and biochemical properties after induction, they did not develop the electrophysiological characteristics of neurons. Thus, these results have suggested that canine BMSCs could have the capacity to differentiate into a neuronal lineage, but the differentiation protocol used may have been insufficient to induce development into functional neurons.

Objective: To measure the effect of cold compress application on tissue temperature in healthy dogs. Animals: 10 healthy mixed-breed dogs. Procedures: Dogs were sedated with hydromorphone (0.1 mg/kg, IV) and diazepam (0.25 mg/kg, IV). Three 24-gauge thermocouple needles were inserted to a depth of 0.5 (superficial), 1.0 (middle), and 1.5 (deep) cm into a shaved, lumbar, epaxial region to measure tissue temperature. Cold (–16.8°C) compresses were applied with gravity dependence for periods of 5, 10, and 20 minutes. Tissue temperature was recorded before compress application and at intervals for up to 80 minutes after application. Control data were collected while dogs received identical sedation but with no cold compress. Results: Mean temperature associated with 5 minutes of application at the superficial depth was significantly decreased, compared with control temperatures. Application for 10 and 20 minutes significantly reduced the temperature at all depths, compared with controls and 5 minutes of application. Twenty minutes of application significantly decreased temperature at only the middle depth, compared with 10 minutes of application. Conclusions and Clinical Relevance: With this method of cold treatment, increasing application time from 10 to 20 minutes caused a further significant temperature change at only the middle tissue depth; however, for maximal cooling, the minimum time of application should be 20 minutes. Possible changes in tissue temperature and adverse effects of application > 20 minutes require further evaluation.

Objective: To evaluate the growth kinetics of a strain of Bifidobacterium pseudocatenulatum (BP) on 4 oligo- or polysaccharides and the effect of feeding a selected probiotic-prebiotic combination on intestinal microbiota in cats. Animals: 10 healthy adult cats. Procedures: Growth kinetics of a strain of cat-origin BP (BP-B82) on fructo-oligosaccharides, galacto-oligosaccharides (GOS), lactitol, or pectins was determined, and the combination of GOS and BP-B82 was selected. Cats received supplemental once-daily feeding of 1% GOS–BP-B82 (10(10) CFUs/d) for 15 days; fecal samples were collected for analysis the day before (day 0) and 1 and 10 days after the feeding period (day 16 and 25, respectively). Results: Compared with the prefeeding value, mean fecal ammonia concentration was significantly lower on days 16 and 25 (288 and 281 μmol/g of fecal dry matter [fDM], respectively, vs 353 μmol/g of fDM); fecal acetic acid concentration was higher on day 16 (171 μmol/g of fDM vs 132 μmol/g of fDM). On day 16, fecal concentrations of lactic, n-valeric, and isovaleric acids (3.61, 1.52, and 3.55 μmol/g of fDM, respectively) were significantly lower than on days 0 (5.08, 18.4, and 6.48 μmol/g of fDM, respectively) and 25 (4.24, 17.3, and 6.17 μmol/g of fDM, respectively). A significant increase in fecal bifidobacteria content was observed on days 16 and 25 (7.98 and 7.52 log10 CFUs/g of fDM, respectively), compared with the prefeeding value (5.63 log10 CFUs/g of fDM). Conclusions and Clinical Relevance: Results suggested that feeding 1% GOS–BP-B82 combination had some positive effects on the intestinal microbiota in cats.

Objective: To characterize equine muscle tissue– and periosteal tissue–derived cells as mesenchymal stem cells (MSCs) and assess their proliferation capacity and osteogenic potential in comparison with bone marrow– and adipose tissue–derived MSCs. Sample: Tissues from 10 equine cadavers. Procedures: 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 adipose tissue from the left subcutaneous region. Mesenchymal stem cells were characterized on the basis of morphology, adherence to plastic, trilineage differentiation, and detection of stem cell surface markers via immunofluorescence and flow cytometry. Mesenchymal stem cells were tested for osteogenic potential with osteocalcin gene expression via real-time PCR assay. Mesenchymal stem cell cultures were counted at 24, 48, 72, and 96 hours to determine tissue-specific MSC proliferative capacity. Results: Equine muscle tissue– and periosteal tissue–derived cells were characterized as MSCs on the basis of spindle-shaped morphology, adherence to plastic, trilineage differentiation, presence of CD44 and CD90 cell surface markers, and nearly complete absence of CD45 and CD34 cell surface markers. Muscle tissue–, periosteal tissue–, and adipose tissue–derived MSCs proliferated significantly faster than did bone marrow–derived MSCs at 72 and 96 hours. Conclusions and Clinical Relevance: Equine muscle and periosteum are sources of MSCs. Equine muscle- and periosteal-derived MSCs have osteogenic potential comparable to that of equine adipose- and bone marrow–derived MSCs, which could make them useful for tissue engineering applications in equine medicine.

Objective: To determine the effects of hypoglossal nerve block and electrical stimulation of the thyrohyoideus muscles on position of the larynx and hyoid apparatus in resting horses. Animals: 16 healthy horses that underwent hypoglossal nerve block and 5 healthy horses that underwent electrical stimulation of the thyrohyoideus muscles. Procedures: Horses underwent bilateral hypoglossal nerve block or electrical stimulation of the thyrohyoideus muscles. Positions of the basihyoid bone, ossified part of the thyroid cartilage, and articulations of the thyrohyoid bones and thyroid cartilage were determined in radiographic images obtained before and after performance of hypoglossal nerve blocks or during thyrohyoideus muscle stimulation. Radiographic images were obtained with the heads of horses in neutral (thyrohyoideus muscle stimulation) or neutral and extended (hypoglossal nerve block) positions. Radiographic images of horses obtained after performance of hypoglossal nerve blocks were also evaluated to detect dorsal displacement of the soft palate. Results: Hypoglossal nerve blocks did not induce significant changes in the positions of evaluated anatomic sites in radiographic images obtained in neutral or extended head positions. Hypoglossal nerve block did not induce dorsal displacement of the soft palate in horses at rest. Bilateral thyrohyoideus muscle stimulation induced significant dorsal movement (mean ± SD change in position, 18.7 ± 6.8 mm) of the ossified part of the thyroid cartilage; rostral movement of evaluated anatomic structures was small and not significant after thyrohyoideus muscle stimulation. Conclusions and Clinical Relevance: Bilateral electrical stimulation of the thyrohyoideus muscles in horses in this study induced dorsal laryngeal movement.

Objective-To evaluate the righting reflex after topical application of a sevoflurane jelly in cane toads (Bufo marinus). Animals-8 cane toads. Procedures-Toads were 6 to 8 months of age and weighed (mean +/- SD) 142.0 +/- 25.2 g. Sevoflurane jelly was applied to the dorsum of each toad at a dose of 25 μL/g in trial 1 and 37.5 μL/g in trial 2. Toads were placed in dorsal recumbency every 30 seconds until loss of the righting reflex. Jelly was then removed by rinsing the toads with tap water. Toads were then left undisturbed in dorsal recumbency until return of the righting reflex. Chamber sevoflurane concentration was measured to determine vaporization. Results-6 of 8 toads in trial 1 and 8 of 8 toads in trial 2 lost the righting reflex. Mean +/- SD time to loss of the reflex was 8.2 +/- 1.3 minutes for trial 1 and 8.3 +/- 0.9 minutes for trial 2; this difference was not significant. Mean +/- SD time to return of the reflex was 25.6 +/- 26.2 minutes for trial 1 and 84.4 +/- 47.2 minutes for trial 2; this difference was significant. Chamber sevoflurane concentration did not change significantly, compared with baseline (time 0) concentration, at any time in trial 1; however, there was a significant change in chamber sevoflurane concentration from baseline (time 0) concentration in trial 2. Chamber sevoflurane concentrations were not significantly different between trial 1 and trial 2 at any time. Mean +/- SD chamber sevoflurane concentration was 0.46 +/- 0.2% for trial 1 and 0.57 +/- 0.28% for trial 2. Conclusions and Clinical Relevance-Sevoflurane jelly applied topically at a dose of 37.5 μL/g induced a more reliable loss of righting reflex and longer recovery time than when applied at a dose of 25 μL/g in cane toads

Objective—To optimize the overall hemostasis potential (OHP) assay for use with canine platelet-poor plasma and determine reference intervals in healthy dogs. Animals—40 healthy dogs. Procedures—Blood was collected from the dogs into citrated tubes, and platlet-poor plasma was obtained. The OHP assay and standard coagulation assays (prothrombin time, activated partial thromboplastin time, and fibrinogen concentration) were performed for each sample. The OHP assay outputs were tested for correlations with results of the standard coagulation assays, age, and sex. Results—Modifications to the published methodology for the OHP assay were required for use with canine plasma, with less coagulation activator (thrombin) and more fibrinolysis activator (tissue plasminogen activator) than used with human plasma. Male dogs had a higher OHP than did females. High fibrinogen concentrations were associated with increases in maximum optical density, OHP, and overall coagulation potential, and reduced prothrombin time was associated with increases in maximum optical density, overall coagulation potential, OHP, and maximum slope. Conclusions and Clinical Relevance—Results supported the use of the OHP assay as an accessible, cost-effective global coagulation assay. Further research is required to determine its clinical application as an alternative to thromboelastography or thrombin generation assays.

Objective-To quantitatively and qualitatively compare electroretinography (ERG) recordings in awake, sedated, and anesthetized dogs. Animals-Six 6-month-old Beagles. Procedures-A brief ERG protocol for dogs was used. Following 1-minute and subsequent 5-minute dark adaptation, mixed rod-cone responses were recorded bilaterally with a handheld multispecies ERG device with dogs in each of 3 states of consciousness: awake, sedated (dexmedetomidine and butorphanol), and anesthetized (atropine and hydromorphone, followed by propofol and midazolam and anesthetic maintenance with isoflurane). Low- and high-frequency noise levels were quantified via Fourier analysis, and the effect of consciousness state on signal amplitude, implicit time, and noise was analyzed via repeated-measures ANOVA. In addition, 13 veterinary ophthalmologists who were unaware of the dogs' consciousness states subjectively graded the ERG recording quality, and scores for each tracing were compared. Results-ERG amplitudes were highest in awake dogs and lowest in anesthetized dogs. Implicit times were shortest in awake dogs and longest in anesthetized dogs. Differences in b-wave amplitudes and a-wave implicit times were significant. Neither low- nor high-frequency noise levels differed significantly among consciousness states. Furthermore, no significant differences were identified among observers' scores assigned to ERG tracings. Conclusions and Clinical Relevance-Anesthesia and sedation resulted in significant attenuation and delay of ERG responses in dogs. Chemical restraint of dogs had no consistently significant effect on low- or high-frequency noise levels or on observer perception of signal quality.