Objective-To determine the efficiency of a novel point-of-care gravitational marrow separator and bone marrow aspiration needle for concentrated bone marrow production and bone marrow-derived mesenchymal stem cell (MSC) separation and assess the effect of repeated bone marrow collections in horses. Animals-8 healthy adult horses. Procedures-Bone marrow aspiration was performed twice (1 month apart) from sternebral bodies with a standard or prototype multidirectional needle. Concentrated bone marrow was obtained by gravitational marrow separation and evaluated for WBC and platelet counts, automated and cytomorphologic cell differential counts, MSCs, and cell viability. Results-Concentrated bone marrow samples obtained with the marrow separator had 5- to 19-fold bone marrow-derived MSC, WBC, and platelet counts, compared with original bone marrow samples. Use of a multidirectional needle increased the frequency of obtaining MSC-richer concentrated bone marrow. Repeating bone marrow aspiration at 1 month yielded greater MSC numbers but slightly lower cell viability after processing. Conclusions and Clinical Relevance-The gravitational bone marrow separator and multidirectional needle were used to effectively harvest bone marrow and improve the quality of concentrated bone marrow. Comparable, or even greater, numbers of bone marrow-derived MSCs were collected by repeated bone marrow aspiration after a 1-month interval from the same aspiration sites. Use of the marrow separator and multidirectional bone marrow aspiration needle can facilitate a 1-step, point-of-care, nonlaboratory method to obtain concentrated bone marrow as a mixture of bone marrow-derived MSCs and growth factors from platelets and plasma.

Objective: To evaluate fecal calprotectin concentrations in healthy dogs and dogs with chronic diarrhea, to identify cutoff values for fecal calprotectin concentrations for use in differentiating dogs with chronic diarrhea and a canine chronic enteropathy clinical activity index (CCECAI) < 12 from dogs with chronic diarrhea and a CCECAI ≥ 12, and to evaluate the association between histologic evidence of intestinal mucosal changes and fecal calprotectin concentrations in dogs with chronic diarrhea. Sample: Fecal samples from 96 adult dogs (27 dogs with chronic diarrhea and 69 healthy control dogs). Procedures: Severity of clinical signs was evaluated on the basis of the CCECAI scoring system. Endoscopy was performed in all dogs with chronic diarrhea, and mucosal biopsy specimens were evaluated histologically. Fecal calprotectin concentration was quantified via radioimmunoassay. Results: Fecal calprotectin concentrations were significantly higher in dogs with chronic diarrhea than in healthy control dogs. Fecal calprotectin concentrations were also significantly higher in dogs with a CCECAI ≥ 12, compared with concentrations for dogs with a CCECAI between 4 and 11. Fecal calprotectin concentrations were significantly higher in dogs with chronic diarrhea associated with histologic lesions, compared with concentrations in control dogs, and were significantly correlated with the severity of histologic intestinal lesions. Among dogs with chronic diarrhea, the best cutoff fecal calprotectin concentration for predicting a CCECAI ≥ 12 was 48.9 μg/g (sensitivity, 53.3%; specificity, 91.7%). Conclusions and Clinical Relevance: Fecal calprotectin may be a useful biomarker in dogs with chronic diarrhea, especially dogs with histologic lesions.

Objective—To determine an efficient method for the collection of semen samples by means of electroejaculation, characterize spermatozoa quality and quantity, and determine the effect of refrigerated storage on motility of spermatozoa obtained from green iguanas (Iguana iguana). Animals—18 adult green iguanas. Procedures—Green iguanas were anesthetized, and semen samples were obtained by means of electroejaculation. Up to 3 series of electrostimulations were performed; the procedure was stopped after a semen sample was obtained. Various semen sample variables were evaluated. Results—Semen samples were obtained from 16 iguanas; most (n = 10) iguanas produced a semen sample after the second series of electrostimulations. Median semen sample volume was 0.05 mL. Mean spermatozoa concentration was 2 69.0 × 10(6) spermatozoa/mL. Median percentage of motile spermatozoa was 78%. The only morphological abnormality of spermatozoa was bent tails (mean percentage in a semen sample, 5.7%). Spermatozoa motility decreased significantly during refrigeration (4°C); median percentage motility after 24, 48, and 72 hours of refrigeration was 60%, 33%, and 0%, respectively. Conclusions and Clinical Relevance—Results of this study suggested electroejaculation can be performed to collect semen samples from green iguanas, characteristics of iguana semen samples are similar to those for semen samples obtained from other reptiles, and motility of iguana spermatozoa decreases during refrigeration within 48 to 72 hours.

Objective-To assess the feasibility of measuring cord dorsum potentials (CDPs) in anesthetized clinically normal dogs after caudal nerve stimulation, determine the intervertebral site of maximum amplitude and best waveform of the CDP, and evaluate the effects of neuromuscular blockade. Animals-8 male and 4 female dogs (age, 1 to 5 years). Procedures-Dogs were anesthetized, and CDPs were recorded via needles placed on the dorsal lamina at intervertebral spaces L1-2 through L7-S1. Caudal nerves were stimulated with monopolar electrodes inserted laterally to the level of the caudal vertebrae. Dogs were tested without and during neuromuscular blockade induced with atracurium besylate. The CDP latency and amplitude were determined from the largest amplitude tracings. Results-CDPs were recorded in 11 of 12 dogs without neuromuscular blockade and in all dogs during neuromuscular blockade. The CDP was largest and most isolated at the L4-5 intervertebral space (3 dogs) or the L5-6 intervertebral space (9 dogs); this site corresponded to the segment of insertion of the first caudal nerve. Onset latencies ranged from 2.0 to 4.7 milliseconds, and there was no effect of neuromuscular blockade on latencies. Amplitudes of the CDPs were highly variable for both experimental conditions. Conclusions and Clinical Relevance-CDPs were recorded from all dogs tested in the study; neuromuscular blockade was not critical for successful CDP recording but reduced muscle artifact. This technique may be useful as a tool to assess the caudal nerve roots in dogs suspected of having compressive lumbosacral disease or myelopathies affecting the lumbar intumescence.

Objective-To compare presumed fatty acid content in natural diets of feral domestic cats (inferred from body fat polyunsatrated fatty acids content) with polyunsaturated fatty acid content of commercial feline extruded diets. Sample-Subcutaneous and intra-abdominal adipose tissue samples (approx 1 g) from previously frozen cadavers of 7 adult feral domestic cats trapped in habitats remote from human activity and triplicate samples (200 g each) of 7 commercial extruded diets representing 68% of market share obtained from retail stores. Procedures-Lipid, triacylglycerol, and phospholipid fractions in adipose tissue samples and ether extracts of diet samples were determined by gas chromatography of methyl esters. Triacylglycerol and phospholipid fractions in the adipose tissue were isolated by thin-layer chromatography. Diet samples were also analyzed for proximate contents. Results-For the adipose tissue samples, with few exceptions, fatty acids fractions varied only moderately with lipid fraction and site from which tissue samples were obtained. Linoleic, α-linolenic, arachidonic, eicosapentaenoic, and docosahexaenoic acid fractions were 15.0% to 28.2%, 4.5% to 18.7%, 0.9% to 5.0%, < 0.1% to 0.2%, and 0.6% to 1.7%, respectively. As inferred from the adipose findings, dietary fractions of docosahexaenoic and α-linolenic acid were significantly greater than those in the commercial feline diets, but those for linoleic and eicosapentaenoic acids were not significantly different. Conclusions and Clinical Relevance-The fatty acid content of commercial extruded feline diets differed from the inferred content of natural feral cat diets, in which dietary n-3 and possibly n-6 polyunsaturated fatty acids were more abundant. The impact of this difference on the health of pet cats is not known.

Objective-To determine the maximum amount of flexion and extension of the carpal, tarsal, metacarpophalangeal, and metatarsophalangeal joints and the percentage duration of the stance and swing phases of the stride for horses walking on an underwater treadmill in various water depths. Animals-9 healthy adult horses. Procedures-Zinc oxide markers were placed on the forelimbs and hind limbs of the horses. Video was recorded of horses walking (0.9 m/s) on an underwater treadmill during baseline conditions (< 1 cm of water) or in various amounts of water (level of the metatarsophalangeal, tarsal, and stifle joints). Maximum amount of joint flexion and extension, range of motion (ROM), and the percentage durations of the stance and swing phases of the stride were determined with 2-D motion analysis software. Results-The ROM was greater for all evaluated joints in any amount of water versus ROM for joints in baseline conditions (primarily because of increases in amount of joint flexion). The greatest ROM for carpal joints was detected in a tarsal joint water depth, for tarsal joints in a stifle joint water depth, and for metacarpophalangeal and metatarsophalangeal joints in metatarsophalangeal and tarsal joint water depths. As water depth increased, the percentage durations of the stance and swing phases of the stride significantly decreased and increased, respectively. Conclusions and Clinical Relevance-Results of this study suggested that exercise on an underwater treadmill is useful for increasing the ROM of various joints of horses during rehabilitation and that the depth of water affects the amount of flexion and extension of joints.

Objective: To determine whether pimobendan has in vitro antithrombotic properties through inhibition of platelets in canine blood samples. Animals: 10 healthy adult dogs. Procedures: Blood samples were collected from each dog into tubes containing hirudin or sodium citrate. Pimobendan was added to blood samples (final concentration, 0.0, 0.01, 0.1, 1.0, or 10.0μM) containing hirudin prior to undergoing collagen- and ADP-induced whole blood impedance aggregometry. Plasma thromboxane concentrations were measured after platelet aggregation. Pimobendan was also added to blood samples (0.0, 0.01, or 10.0μM) containing sodium citrate prior to thromboelastographic evaluation. Results: Compared with findings for 0.0μM pimobendan, composite platelet aggregation (area under the curve [AUC]) and maximal platelet aggregation (aggregation units [AUs]) at 10.0μM pimobendan were significantly decreased for collagen-induced aggregation (AUC, 349.7 ± 58.4 vs 285.1 ± 72.2; maximal platelet aggregation, 196.2 ± 25.8 AUs vs 161.5 ± 38.0 AUs), and the AUC and velocity of aggregation at 10.0μM pimobendan were significantly decreased for ADP-induced aggregation (AUC, 268.5 ± 35.1 vs 213.4 ± 77.2; velocity of aggregation, 15.7 ± 2.9 AUs/min vs 11.8 ± 3.5 AUs/min). Pimobendan had no significant effect on plasma thromboxane concentration or thromboelastographic variables, regardless of concentration. Conclusions and Clinical Relevance: In vitro, pimobendan had an antiplatelet effect in canine blood samples at a concentration 1,000-fold higher than that clinically achievable. These antiplatelet properties do not appear to contribute to the positive clinical profile of the drug in dogs. Pimobendan administration would not appear to confer a risk for bleeding and does not have to be avoided in dogs with thrombocytopenia or those concurrently receiving antiplatelet drugs.

Objective: To evaluate the influence of 3 anesthetic protocols and multiples of minimum alveolar concentration (MAC) on heart rate variability (HRV) with and without nociceptive stimulation in dogs. Animals: 6 healthy adult Beagles. Procedures: Each dog was anesthetized 3 times: with isoflurane alone, with isoflurane and a constant rate infusion of dexmedetomidine (IsoD; 3 μg/kg/h, IV), and with isoflurane and a constant rate infusion of remifentanil (IsoR; 18 μg/kg/h, IV). Individual MAC was determined via supramaximal electrical stimulation. Sinus rhythm–derived intervals between 2 adjacent R-R intervals were exported from ECG recordings. Selected HRV time and frequency domain variables were obtained (at 2-minute intervals) and analyzed offline with signed rank tests before and after stimulation at 0.75, 1.0, and 1. 5 MAC for each anesthetic session. Results: The isoflurane session had the overall lowest prestimulation SDNN (SD of all R-R intervals) values. Prestimulation SDNN values decreased significantly with increasing MAC in all sessions. For the IsoD session, SDNN (milliseconds) or high-frequency power (milliseconds2) was inversely correlated with MAC (Spearman rank correlation coefficient for both variables, −0.77). In the isoflurane and IsoR sessions, heart rate increased significantly after stimulation. In the IsoD session, poststimulation SDNN was increased significantly, compared with prestimulation values, at 0.75 and 1.0 MAC. Conclusions and Clinical Relevance: On the basis of SDNN and high-frequency power values, anesthetic levels between 0.75 and 1.5 MAC within the same anesthetic protocol could be differentiated, but with a large overlap among protocols. Usefulness of standard HRV variables for assessment of anesthetic depth and nociception in dogs is questionable.

Objective: To evaluate first-intention healing of CO2 laser, 4.0-MHz radiowave radiosurgery (RWRS), and scalpel incisions in ball pythons (Python regius). Animals: 6 healthy adult ball pythons. Procedures: A skin biopsy sample was collected, and 2-cm skin incisions (4/modality) were made in each snake under anesthesia and closed with surgical staples on day 0. Incision sites were grossly evaluated and scored daily. One skin biopsy sample per incision type per snake was obtained on days 2, 7, 14, and 30. Necrotic and fibroplastic tissue was measured in histologic sections; samples were assessed and scored for total inflammation, histologic response (based on the measurement of necrotic and fibroplastic tissues and total inflammation score), and other variables. Frequency distributions of gross and histologic variables associated with wound healing were calculated. Results: Gross wound scores were significantly greater (indicating greater separation of wound edges) for laser incisions than for RWRS and scalpel incisions at all evaluated time points. Necrosis was significantly greater in laser and RWRS incisions than in scalpel incision sites on days 2 and 14 and days 2 and 7, respectively; fibroplasia was significantly greater in laser than in scalpel incision sites on day 30. Histologic response scores were significantly lower for scalpel than for other incision modalities on days 2, 14, and 30. Conclusions and Clinical Relevance: In snakes, skin incisions made with a scalpel generally had less necrotic tissue than did CO2 laser and RWRS incisions. Comparison of the 3 modalities on the basis of histologic response scores indicated that use of a scalpel was preferable, followed by RWRS and then laser.

Objective: To determine 2-deoxy-2-fluoro (fluorine 18)-d-glucose (18FDG) biodistribution in the coelom of bald eagles (Haliaeetus leucocephalus). Animals: 8 healthy adult bald eagles. Procedures: For each eagle, whole-body transmission noncontrast CT, 60-minute dynamic positron emission tomography (PET) of the celomic cavity (immediately after 18FDG injection), whole-body static PET 60 minutes after 18FDG injection, and whole-body contrast CT with iohexol were performed. After reconstruction, images were analyzed. Regions of interest were drawn over the ventricular myocardium, liver, spleen, proventriculus, cloaca, kidneys, and lungs on dynamic and static PET images. Standardized uptake values were calculated. Results: Kidneys had the most intense 18FDG uptake, followed by cloaca and intestinal tract; liver activity was mild and slightly more intense than that of the spleen; proventricular activity was always present, whereas little to no activity was identified in the wall of the ventriculus. Activity in the myocardium was present in all birds but varied in intensity among birds. The lungs had no visibly discernible activity. Mean ± SD standardized uptake values calculated with representative regions of interest at 60 minutes were as follows: myocardium, 1. 6 ± 0.2 (transverse plane) and 1.3 ± 0.3 (sagittal plane); liver, 1.1 ± 0.1; spleen, 0.9 ± 0.1; proventriculus, 1.0 ± 0.1; cloaca, 4.4 ± 2.7; right kidney, 17.3 ± 1.0; left kidney, 17.6 ± 0.3; and right and left lungs (each), 0.3 ± 0.02. Conclusions and Clinical Relevance: The study established the biodistribution of 18FDG in adult eagles, providing a baseline for clinical investigation and future research.