DEVELOPMENT OF A HEMOLYMPH COLLECTION TECHNIQUE AND REPORT OF BIOCHEMICAL PARAMETERS OF HEMOLYMPH IN A MANAGED POPULATION OF FEMALE THORNY DEVIL STICK INSECTS (EURYCANTHA CALCARATA).

Insects are increasingly common in households, zoological collections, research, and food industries. Increased knowledge of insect health parameters is necessary to ensure proper welfare. The study goal was to develop a hemolymph collection technique and report hemolymph serum biochemical parameters for the thorny devil stick insect (Eurycantha calcarata). Clinically healthy adult stick insects (5 males and 14 females, 15.1-24.7 g) were enrolled. Four collection techniques were evaluated. Hemolymph collection was unsuccessful in males, but was successful in females by using a single technique. The insect was manually restrained in an elevated position and an 18-ga × 2.54-cm needle was used to puncture the membrane just caudal to the third pair of legs. With the puncture site directed ventrally, ≤1 ml of hemolymph was collected via gravity and allowed to visibly clot. The sample was then centrifuged, and the serum was separated. Serum samples were individually analyzed (Avian/Reptilian Profile Plus, VetScan VS2, Abaxis, Inc, Union City, CA 94587, USA). Fourteen samples (0.2-1.0 ml) were collected from 14 females. Median (minimum-maximum) parameters included the following: aspartate aminotransferase 12 (0-45) U/L, creatinine kinase 25 (0-76) U/L, uric acid 7.5 (3.1-13.7) mg/dl, glucose 12 (8-22) mg/dl, calcium 18.6 (17.2-19.4) mg/dl, phosphorus 15.0 (n = 1) or >30.0 (n = 13) mg/dL, total protein 2.7 (1.6-2.9) g/dL, albumin 0.9 (0.2-1.2) g/dL, globulin 1.7 (1.6-1.8) g/dL (n = 6) or not quantified (n = 8), potassium 10.6 (9.0-11.8) mmol/L, sodium < 100 mmol/L, and bile acids 0 lmol/L. This is the first report of biochemistry parameters in clinically healthy female stick insects. Larger sample sizes are needed to establish statistically valid reference ranges. Hemolymph collection techniques for male stick insects warrant further investigation.

INVESTIGATION OF A POINT-OF-CARE VISCOELASTIC COAGULATION MONITOR AND ITS COMPARISON TO THROMBOELASTOGRAPHY IN CLINICALLY HEALTHY AFRICAN ELEPHANTS (LOXODONTA AFRICANA).

Elephant endotheliotropic herpesvirus (EEHV) can induce fatal hemorrhagic disease (HD) in African elephants (Loxodonta africana). Once clinical signs develop, progression is rapid, even with aggressive treatment. There is a critical need to develop point-of-care diagnostic tests to aid in identification of EEHV-HD prior to the onset of overt clinical signs. Study objectives were to investigate a novel, point-of-care viscoelastic coagulation monitor (VCM Vet), compare the results to thromboelastography (TEG), and report traditional hemostatic analytes in adult African elephants. Whole blood was collected from seven clinically healthy elephants (four females and three males, 18-47 yr) and analyzed in duplicate via VCM Vet and kaolin-activated TEG 1-3 and 30 min following collection, respectively. Separated plasma was frozen for ancillary coagulation testing. Both analyses generated quantifiable clotting reactions with variables (median [range]) describing clot formation rate (VCM Vet, clot time = 682 s [530-987 s], clot formation time = 244 s [186-744 s], Alpha = 40° [14-47°]; TEG, reaction time = 6.2 min [3.7-11.8 min], kinetic time = 1.3 min [0.9-2.6 min], Alpha = 70° [57-77°]), clot strength (VCM Vet, maximum clot formation = 34 units [20-45 units]; TEG, maximum amplitude = 75 mm [69-80 mm], shear elastic modulus strength = 14.7 Kdynes/s [11.3-19.5 Kdynes/s]), and clot lysis (VCM Vet, lysis index at 30 min = 100% [100-99%], lysis index at 45 min = 98% [95-100%]; TEG, lysis index at 30 min = 0% [0-0.4%], lysis index at 60 min = 1.4% [0-2.6%]) recorded. Additional testing (median [range]) included D-dimer concentration (33 ng/ml [28-94 ng/ml]), prothrombin time (12.4 s [12.2-13.2 s]), activated partial thromboplastin time (17.2 s [14.2-18.8 s]), and fibrinogen concentration (297 [282-383] mg/dL). Tracings generated by VCM Vet and TEG were clinically similar, and there was visual agreement and minimal difference between quantitative variables for duplicate tests. VCM Vet is a promising, user-friendly tool for use in identification and management of coagulopathies in African elephants.

POTASSIUM CHLORIDE ADMINISTERED VIA FOUR ROUTES FOR EUTHANASIA OF ANESTHETIZED GOLDFISH (CARASSIUS AURATUS).

Immersion overdose in tricaine methanesulfonate (MS-222) is ineffective for euthanasia of adult goldfish (Carassius auratus), and investigation of alternative techniques is warranted. This study evaluated potassium chloride (KCl) administered via four routes for goldfish euthanasia. Thirty clinically healthy adult goldfish (17.2-41.9 g) were anesthetized via immersion in buffered MS-222 (300 mg/L) for 5 min beyond anesthetic induction. Fish were then randomly administered one of five treatments: KCl (333 mg/mL, 4.5 mmol/mL, 4.5 mEq/mL) via intracardiac injection at 10 mEq/kg (IC), intracoelomic injection (ICe) or bilateral topical delivery over the gill filaments (T) at 100 mEq/kg, or 90-min immersion at 4,500 mEq/L (saturated solution) dissolved in induction solution (water [W]), or no treatment (control [C]). Following treatment, all fish were moved to anesthetic-free freshwater. Serial heart rates were collected via Doppler device until sound cessation or recovery. Median (range) time to perform treatments was 5.3 (1.2-8.0) min, 3 (3-3) s, and 10 (10-10) s in IC, ICe, and T, respectively. Doppler cessation occurred in 6/6, 6/6, 6/6, 6/6, and 0/6 fish in median (range) times of 3 (0-210), 18 (10-45), 118 (90-390), and 150 (60-240) min in IC, ICe, T, W, and C, respectively; this was significantly different between groups (P = 0.002). Following or during treatment, 1/6, 2/6, 6/6, and 4/6 fish in IC, ICe, T, and W, respectively, exhibited intermittent transient jerking movements. Median (range) time to recovery in C was 5.5 (3.5-6.5) min. All administration routes were effective, but time to Doppler cessation varied and transient movements were noted. Preliminary follow-up research determined that return to MS-222 induction solution following ICe KCl treatment abolished movements. ICe KCl at 100 mEq/kg was technically simple and resulted in Doppler sound cessation in a clinically relevant time frame in 6/6 anesthetized goldfish. Return to MS-222 immersion following KCl administration is recommended.

EVALUATION OF INTRACARDIAC ADMINISTRATION OF POTASSIUM CHLORIDE, IVERMECTIN, OR LIDOCAINE HYDROCHLORIDE FOR EUTHANASIA OF ANESTHETIZED BLUE CRABS (CALLINECTES SAPIDUS).

Methods to anesthetize and euthanize aquatic invertebrates have proven unreliable in decapods; thus studies to optimize euthanasia techniques for crustaceans are needed. Study objectives were to evaluate efficacy of intracardiac potassium chloride (KCl), ivermectin, or lidocaine hydrochloride (HCl) for euthanasia of anesthetized blue crabs (Callinectes sapidus). Twenty adult male crabs (n = 5/group) were immersed in 500 mg/L eugenol for 5 min beyond loss of the righting reflex and then randomly administered intracardiac 10 mEq/kg KCl (333 mg/mL), 5 mg/kg ivermectin (10 mg/ml), 100 mg/kg lidocaine hydrochloride (HCl) (20 mg/ml), or 5 ml/kg saline (0.9%). Serial heart rate assessments were made using a Doppler probe placed over the dorsum, and times to loss of righting reflex, Doppler sound cessation, and/or recovery were recorded. Median (range) time to loss of righting reflex was 32 (17-57) min. One crab in all groups, except lidocaine HCl, had no detectable Doppler sounds prior to injection. In the remaining crabs, Doppler sound cessation occurred in 4/4, 4/4, 4/5, and 0/4 crabs administered KCl, ivermectin, lidocaine HCl, and saline, respectively. Median (range) time to Doppler sound cessation was 30 (0-55), 18 (16-28), and 50 (0-90) s in KCl, ivermectin, and lidocaine HCl groups, respectively. Tonic limb movements were observed in 5/5 KCl-treated crabs. Median (range) time to recovery was 180 (115-345) and 300 m in four saline-treated crabs and one lidocaine HCl-treated crab, respectively. Intracardiac KCl at 10 mEq/kg and ivermectin at 5 mg/kg were effective, rapid methods for euthanasia of anesthetized blue crabs.

EVALUATION OF OSCILLOMETRIC BLOOD PRESSURE MEASUREMENT USING A FINGER CUFF IN ANESTHETIZED CHIMPANZEES (PAN TROGLODYTES).

Cardiovascular disease is common among chimpanzees (Pan troglodytes), and serial blood pressure monitoring in conscious animals may improve disease surveillance and guide hypertension treatment strategies. The objective of this study was to compare the accuracy of a noninvasive, oscillometric blood pressure monitor using a finger blood pressure cuff with invasively measured blood pressure in anesthetized chimpanzees. Twelve chimpanzees were anesthetized with tiletamine-zolazepam intramuscularly, intubated, and maintained on inhaled isoflurane to effect. Blood pressure measurements, which included systolic arterial pressure (SAP), mean arterial pressure (MAP), and diastolic arterial pressure (DAP), were collected simultaneously from an oscillometric blood pressure cuff placed on a forelimb digit (FBP) and a direct arterial catheter (IBP) every 5-10 min while anesthetized. One hundred paired samples were collected, and results were compared using Bland-Altman plots and analysis. FBP showed good agreement with IBP for SAP, MAP, and DAP but consistently overestimated values compared with IBP. FBP may be useful for serial blood pressure monitoring in conscious chimpanzees.

Point-of-care viscoelastic coagulation assessment in healthy dogs during the perianesthetic period.

BACKGROUND: The viscoelastic coagulation monitor (VCM Vet) is a novel, portable device that provides a global assessment of hemostasis. The study aims were to evaluate serial viscoelastic analysis during the perianesthetic period in healthy dogs and to compare the agreement between two VCM Vet devices. Twenty healthy dogs undergoing orthopedic surgery were enrolled. Whole blood samples were collected from an intravenous catheter at four time points: baseline, 15 min after premedication, 60 min after inhalant initiation, and 60 min after inhalant termination. Viscoelastic tests were performed in duplicate on different devices, providing: clot time (CT; seconds), clot formation time (CFT; seconds), alpha angle (α; degrees), amplitude (units) at 10 (A10) and 20 (A20) minutes post clot time, maximum clot firmness (MCF; units), and lysis index (%) at 30 (Li30) and 45 (Li45) minutes post maximum clot formation. RESULTS: One hundred sixty samples were analyzed. The speed of CT and CFT significantly decreased an average of 25.5 s (95% confidence interval [CI]15.9-35.0) and 6.9 s (95% CI 3.1-10.7) per time point, respectively. There were no significant changes in clot strength or lysis variables. The Bland-Altman style plot shows an acceptable rate of agreement for all variables with intra-class correlation ranging from 0.64-0.94. CONCLUSION: The rate of clot formation (CT and CFT) decreased over the perianesthetic period in healthy dogs undergoing surgery. These changes were small and occurred without changes in clot strength or fibrinolysis rate, thus were not clinically relevant. There was clinically acceptable consistency between devices.

Effects of Intramuscular Alfaxalone and Midazolam Compared With Midazolam and Butorphanol in Rhode Island Red Hens (Gallus gallus domesticus).

Chickens (Gallus gallus domesticus) often undergo veterinary procedures requiring sedation; however, there is little published research evaluating the efficacy of sedation protocols in this species. The objective of this study was to assess the effects of intramuscular alfaxalone and midazolam compared with intramuscular butorphanol and midazolam in chickens. In a complete crossover study, 11 healthy adult hens were randomly administered midazolam 2.5 mg/kg IM combined with either alfaxalone 15 mg/kg IM (AM, n = 11) or butorphanol 3 mg/kg IM (BM, n = 11), with a 35-day washout period between groups. Time to first effects, recumbency, standing, and recovery were recorded. Physiologic parameters and sedation scores were recorded every 5 minutes by 2 blinded investigators. Fifteen minutes after injection, positioning for sham whole body radiographs was attempted. At 30 minutes, flumazenil 0.05 mg/kg IM was administered to all hens. Peak total sedation score was significantly higher for AM compared with BM (P < 0.001). Mean ± SD or median (range) time to initial effects, recumbency, standing, and recovery in AM and BM were 1.9 ± 0.6 and 2.6 ± 0.9 (P = 0.02), 3.5 (1.6-7.6) and 4.8 (2.2-13.0) (P = 0.10), 40.3 (28.0-77.8) and 33.2 (5.2-41.3) (P = 0.15), and 71.2 (45.7-202.3) and 39.9 (35.9-45.9) minutes (P = 0.05), respectively. Radiographic positioning was successful in 6 of 11 (54.5%) and 0 of 11 (0%) birds in the AM and BM groups at 15 minutes, respectively. Heart and respiratory rates remained within acceptable clinical limits for all birds. Intramuscular AM resulted in significantly faster onset of sedative effects, significantly longer duration of recumbency, significantly higher peak sedation, and improved success of radiographic positioning compared with intramuscular BM. Intramuscular AM produces clinically effective sedation in chickens without clinically significant cardiorespiratory effects.

SEDATIVE AND CARDIORESPIRATORY EFFECTS OF INTRAMUSCULAR ALFAXALONE AND BUTORPHANOL AT TWO DOSAGES IN FERRETS (MUSTELA PUTORIUS FURO).

Veterinary care of ferrets often requires chemical restraint. This study hypothesized that IM alfaxalone and butorphanol would result in clinically useful sedation without clinically relevant cardiorespiratory effects. Twelve healthy 15-mo-old ferrets of equal sexes weighing 0.75 to 1.66 kg were enrolled. Using a prospective, blinded design, ferrets randomly received either IM alfaxalone 2.5 mg/kg and butorphanol 0.2 mg/kg (low dose [LD]) or IM alfaxalone 5 mg/kg and butorphanol 0.2 mg/kg (high dose [HD]) (n = 6/group). Sedation times and induction and recovery scores were recorded by a blinded observer. Anesthetic monitor placement was attempted in all recumbent ferrets, and physiologic parameters and reflexes were recorded every 5 min until return of spontaneous movement. Data were assessed for normality using a Shapiro-Wilk normality test and analyzed by two-sample t test or Mann-Whitney U test; one ferret in HD was excluded. Ferrets in LD and HD exhibited moderate and marked sedation, with one of six and four of five ferrets tolerating monitor placement, respectively. Mean ± SD time to first effects, recumbency, and recovery in LD and HD was 2.30 ± 1.13 and 2.054 ± 1.12 (P = 0.7240), 2.87 ± 1.25 and 2.72 ± 1.41 (P = 0.8529), and 65.43 ± 32.43 and 52.30 ± 13.19 (P = 0.4212), respectively. Median (range) duration of recumbency in LD and HD was 31.12 (25.58-115.72) and 35.47 (28.27-44.42) min (P = 0.3290), respectively. Among monitored ferrets, transient mild hypotension and hypoxemia were observed. Intramuscular alfaxalone 5 mg/kg with butorphanol 0.2 mg/kg provided clinically useful sedation in ferrets with mild transient cardiorespiratory derangements.

Ex vivo comparison of the effect of storage temperature on canine intestinal leakage pressures.

OBJECTIVE: To determine the effect of storage temperature on cadaveric small intestinal leakage pressures after enterotomy. STUDY DESIGN: Experimental ex vivo study. ANIMALS: Grossly normal jejunal segments from four canine cadavers. METHODS: Thirty-six jejunal segments (n = 12 segments/group) were harvested immediately after euthanasia and assigned to a fresh group (tested within 4 hours), chilled group (stored for 24 hours at 4°C before testing), or freeze-thaw group (frozen at -20°C for 7 days and thawed at 21°C for 6 hours before testing). A 2-cm antimesenteric enterotomy was performed and repaired with 4-0 monofilament suture in a simple-continuous pattern. Initial leakage pressure (ILP), maximal intraluminal pressure (MIP), and leakage location were recorded, with testing performed at room temperature. RESULTS: Mean ± SD ILP for fresh, chilled, and frozen-thawed specimens was 52.9 ± 8.4, 51.8 ± 11.9 and 29.8 ± 4.4 mm Hg, respectively. There was a difference in ILP among groups (P < .003), with freeze-thaw samples demonstrating lower ILP compared with other groups. There was no difference in MIP between groups (P = .186) There was a difference in leakage location among groups (P = .004), with the majority of chilled and freeze-thaw samples leaking at the suture holes compared with the incisional line in fresh samples. CONCLUSION: Freezing and subsequent thawing prior to specimen testing reduced ILP compared with use of fresh and chilled specimens but did not affect MIP among experimental groups. CLINICAL SIGNIFICANCE: Cadaveric canine intestinal specimens tested immediately after collection or after chilling for 24 hours should be recommended for ex vivo burst pressure assessment in dogs. Additional studies to evaluate loss in testing viability of chilled intestinal specimens are warranted to help govern experimental methodologies.

EVALUATION OF POTASSIUM CHLORIDE ADMINISTERED VIA THREE ROUTES FOR EUTHANASIA OF ANESTHETIZED KOI (CYPRINUS CARPIO ).

Immersion in tricaine methanesulfonate (MS-222) is insufficient for euthanasia in at least one species of fish. The current study investigated the effectiveness of potassium chloride (KCl) to euthanize anesthetized koi (Cyprinus carpio). Twenty-eight healthy koi were anesthetized via immersion in 500 mg/L of buffered MS-222 for 10-12 min, manually removed to room air, and randomly administered 10 mEq/kg KCl (333 mg/ml) via one of three routes-intracardiac injection (IC) (n = 7), intracoelomic injection (ICe) (n = 7), or topical instillment over the gill filaments bilaterally (T) (n = 7)-or received no treatment (control, C) (n = 7). A Doppler ultrasonic flow detector was placed over the heart, and sounds were assessed continuously from immediately prior to treatment until 5 min posttreatment and every 5 min thereafter until Doppler sound cessation, resumption of operculation, or 30 min. Time to Doppler sound cessation or resumption of operculation was recorded. Doppler sound cessation occurred in 7/7 fish in IC (median 0.08, range 0-2.75 min) and 1/7 fish in T (10 min). In T, ICe, and C, 6/7, 7/7, and 7/7 fish, respectively, maintained Doppler sounds to 30 min. All fish in ICe (7/7) and C (7/7) resumed operculation with median (range) times of 22 (7-30) min and 16 (9-29) min from treatment, respectively. Intracardiac KCl at 10 mEq/kg rapidly ceases Doppler sounds and is a successful technique for euthanasia of anesthetized koi. Intracoelomic and topical KCl at 10 mEq/kg were not effective for euthanasia of koi.

OBJECTIVE GAIT ANALYSIS IN HUMBOLDT PENGUINS (SPHENISCUS HUMBOLDTI) USING A PRESSURE-SENSITIVE WALKWAY.

Assessment of pododermatitis, osteoarthritis, and other causes of lameness in penguins can be challenging. Subjective gait analysis using visual observation and response to analgesic therapy can be affected by observer variation and caregiver placebo bias. A pressure-sensitive walkway (PSW), however, allows for objective gait analysis and assessment of analgesic therapeutic response. In this study, a 3-m-long PSW was used to analyze gait in 21 adult Humboldt penguins (Spheniscus humboldti). Medical record reviews and comprehensive examinations were performed on all penguins; five penguins were considered abnormal, with either right-sided (n = 3) or bilateral historical lameness-causing disease (n = 2) and were analyzed separately from the normal data set. All penguins walked across the PSW four times and gait parameters (step and stride distances and velocities, maximum force, impulse, and peak pressure) were calculated for each foot in each penguin. Statistical comparisons were made between right and left feet, sexes, and normal and abnormal penguins for each gait parameter. Among normal penguins, there were no significant differences between feet or sex. Left step width was shorter in abnormal penguins than that of normal penguins. Study results established baseline values for Humboldt penguins. This will allow objective monitoring of progression and response to therapy in penguin lameness cases, both current and future. The data also provide a foundation to compare gait parameters with other penguin populations and species.

IMMERSION IN TRICAINE METHANESULFONATE (MS-222) IS NOT SUFFICIENT FOR EUTHANASIA OF SMOKEY JUNGLE FROGS (LEPTODACTYLUS PENTADACTYLUS).

Although tricaine methanesulfonate (MS-222) immersion has historically been standard of care for fish and anuran euthanasia, recent research has proven it insufficient for euthanasia of goldfish. To assess appropriateness for humane euthanasia of anurans, this study evaluated the efficacy of MS-222 in Smokey Jungle Frogs (Leptodactylus pentadactylus). Eighteen frogs (21-33 g) were exposed to one of three MS-222 concentrations via partial immersion: 2.5 g/L for 90 min (M2.5/90), 5 g/L for 60 min (M5/60), or 10 g/L for 60 min (M10/60). Physiologic parameters and times to loss of spontaneous movement, righting reflex, and noxious stimulus response were recorded. Following exposure, frogs were rinsed with dechlorinated water, and time to cessation of heart beat was recorded. Survival in M2.5/90, M5/60, and M10/60 was one of six, zero of six, and zero of six, respectively. In M2.5/90, three of six frogs had continued purposeful, spontaneous movement throughout exposure. In M5/60 and M10/60, median (range) time to initial loss of movement was 14.3 (5.5-30.0) and 7.6 (4.8-19.7) min, respectively. Twelve of 18 frogs among all groups demonstrated a median (range) of two (one to six) episodes of regained consciousness with purposeful, spontaneous movement following loss of noxious stimulus response. Median (range) time to heart beat cessation in M2.5/90, M5/60, and M10/60 was 150 (135-210), 157.5 (60-225), and 90 (75-210) min, respectively. Although death was achieved in 17 of 18 frogs, given the repeated events of regained consciousness, MS-222 immersion when used at concentrations ≤10 g/L did not result in rapid and distress-free death and is not sufficient for humane euthanasia in this species.

INHALANT ANESTHETIC RECOVERY FOLLOWING INTRAMUSCULAR EPINEPHRINE IN THE LOGGERHEAD SEA TURTLE ( CARETTA CARETTA).

Prolonged anesthetic recovery time is a common complication of chelonian inhalant anesthesia and may be exacerbated by right-to-left intracardiac shunting of blood. Epinephrine may decrease intracardiac shunting, which may shorten anesthetic recovery time. The study objective was to assess inhalant anesthetic recovery time following intramuscular epinephrine compared with saline in the loggerhead sea turtle ( Caretta caretta). With the use of a prospective, randomized, blinded, crossover design with a 1-wk washout period, six turtles were anesthetized with intravenous (IV) alfaxalone 3 mg/kg, orotracheally intubated, manually ventilated with 3.5% isoflurane inhalant in 100% oxygen for 90 min, and administered either intramuscular (IM) epinephrine 0.1 mg/kg or IM saline 0.1 ml/kg. Isoflurane administration was immediately discontinued and turtles were manually ventilated with room air until extubation. Physiologic variables, sedation scores, end-tidal carbon dioxide (ETCO2) and isoflurane (ETISO) concentrations, time to first movement, and time to extubation were recorded and two-time-point venous blood gas analyses performed. Data were compared with the use of paired t-tests and repeated-measures analyses of variance (ANOVA) ( P < 0.05). No morbidity, mortality, or adverse events occurred. ETCO2 and ETISO did not significantly change over time during the isoflurane delivery period ( P = 0.990). Mean time to first movement was significantly faster following epinephrine (69.24 ± 12.28 min) compared with saline (87.71 ± 27.05 min, P = 0.047). Although differences were not statistically significant ( P = 0.133), time to extubation was at least 30 min faster (31-123 min) in 4/6 turtles following epinephrine compared with saline. Intramuscular epinephrine significantly reduces time to first movement during isoflurane anesthetic recovery in loggerhead sea turtles.

OSTEOCHONDRAL AUTOGRAFT TRANSFER FOR TREATMENT OF STIFLE OSTEOCHONDRITIS DISSECANS IN TWO RELATED SNOW LEOPARDS ( PANTHERA UNCIA).

Two related female snow leopards ( Panthera uncia) were diagnosed with osteochondritis dissecans of the lateral femoral condyle via radiographs and computed tomography. Lesions were either bilateral (Patient 1) or unilateral (Patient 2). Surgical treatment was performed using an osteochondral autograft transfer system following cartilage flap removal. After a 6-wk recovery period, both animals had a markedly improved gait with mild pelvic limb stiffness. Patient 1 developed intermittent lameness 9 mo postsurgery and Patient 2 had mild lameness 3 yr postsurgery. Because of recurrent lameness, both animals were treated with platelet-rich plasma intra-articular injections and oral gabapentin (Patient 1) or oral meloxicam (Patient 2). Full incorporation of autografts was confirmed with computed tomography (both patients) and arthroscopy (Patient 1 only). Osteochondritis dissecans has been previously reported in snow leopards and these cases represent the first reported use of osteochondral autograft repair and platelet-rich plasma to treat the condition.

Comparison of the Sedative Effects of Alfaxalone and Butorphanol-Midazolam Administered Intramuscularly in Budgerigars (Melopsittacus undulatus).

Intramuscular (IM) administration of either alfaxalone or butorphanol with midazolam was evaluated for sedation and effects on heart and respiratory rates in budgerigars (Melopsittacus undulatus). Twenty adult budgerigars were randomly assigned to receive 1 of 2 treatments: alfaxalone at 15 mg/kg IM or butorphanol and midazolam at 2.5 mg/kg IM and 1.25 mg/kg IM, respectively. Baseline heart and respiratory rates and sedation score were collected and, after injection, time to initial effects and initial recumbency were recorded. Five minutes after injection, birds were assessed for a righting reflex, and, if absent, response to noxious stimulus was assessed by hemostat clamp on the first digit. Birds then underwent a standardized diagnostic evaluation of radiographs, jugular venipuncture, and physical examination. At the end of the physical examination, a sedation score was assigned, and birds were placed in dorsal recumbency for recovery. Times from injection to initial arousal, final recumbency, standing, and full recovery were recorded. Although time to onset of sedation was the same between groups, alfaxalone produced a shorter (P = .04) and more consistent duration of sedation (mean ± SD: 27.5 ± 5.9 minutes) compared with butorphanol-midazolam (72.0 ± 56.9 minutes). Only 3 of 10 birds receiving butorphanol-midazolam were recumbent by 5 minutes compared with 10 of 10 birds receiving alfaxalone. Radiographs were successfully obtained in 9 of 10 and 7 of 10 birds administered alfaxalone and butorphanol-midazolam, respectively. No adverse effects were observed in any bird. Intramuscular alfaxalone produces safe, effective, and reliable sedation in budgerigars and is a more consistent and shorter duration alternative to intramuscular butorphanol-midazolam.

PROPOFOL AS AN IMMERSION ANESTHETIC AND IN A MINIMUM ANESTHETIC CONCENTRATION (MAC) REDUCTION MODEL IN GOLDFISH (CARASSIUS AURATUS).

Propofol is a novel immersion anesthetic in goldfish ( Carassius auratus ). Objectives were to characterize propofol as an anesthetic and assess its suitability in a minimum anesthetic concentration (MAC) reduction model. Using a crossover design, eight goldfish were submerged in 1, 5, or 10 mg/L propofol. Data included induction time, recovery time, heart rate, opercular rate, and response to supramaximal stimulation. Baseline MAC (Dixon's up-and-down method) was determined, and 15 fish were anesthetized with propofol on 4 consecutive days with MAC determination on the fifth day, weekly, for 1 mo. Using a crossover design, MAC of propofol (n = 15) was determined 1 hr following administration of i.m. butorphanol 0.05, 0.5, and 1 mg/kg, dexmedetomidine 0.01, 0.02, and 0.04 mg/kg, ketoprofen 0.5, 1, and 2 mg/kg, morphine 5, 10, and 15 mg/kg, or saline 1 ml/kg. Comparisons were performed with Wilcoxon signed-rank tests (P < 0.05) and Tango's score confidence interval. Propofol at 1 mg/L did not produce anesthesia. Induction time with 10 mg/L (112, 84-166 s) was faster than 5 mg/L (233, 150-289 s; P = 0.0078). Heart and opercular rates for 5 and 10 mg/L were 36 (24-72) beats/min, 58 (44-68) operculations/min and 39 (20-48) beats/min, 57 (48-80) operculations/min, respectively. Recovery time was 249 (143-396) s and 299 (117-886) s with 5 and 10 mg/L, respectively. Response to supramaximal stimulation was not significantly different with 5 mg/L (1/8) compared with 10 mg/L (0/8). Baseline and weekly MAC following daily exposure was 8.4 and 9.0, 8.1, 8.1, and 8.7 mg/L, respectively. MAC reduction was no more than 8% following any drug or dosage. Propofol at 5 and 10 mg/L produced anesthesia, and anesthetic needs were similar following repeated exposure. Propofol was not suitable to test MAC reduction in goldfish in this study.

Popliteal sinus venipuncture in anurans.

OBJECTIVE: To demonstrate an underutilized, reliable, and technically simple anuran venipuncture technique by use of the popliteal sinus. ANIMALS: Medium to large (> 30 g) anuran species. METHODS: The animal is manually restrained using an encircling grip around the pelvic girdle. With the ventrum facing the phlebotomist, the phlebotomist grasps the distal hind limb and gently extends it. If necessary, the medial stifle can be rinsed with sterile water or saline to remove gross debris. Using a caudomedial approach, a 25- to 27-G, 0.5-inch (1.27-cm) needle and attached syringe is advanced toward the stifle at an approximately 45° angle while maintaining negative pressure. If lymphatic fluid or a visibly lymphatic fluid-contaminated blood sample is obtained, the sample is discarded and the attempt is repeated with a new needle and syringe. Collection volume should not exceed 1% of body weight in systemically healthy animals. In compromised animals, a reduction in collection volume should be considered (eg, ≤ 0.5% of body weight). While the phlebotomist may have an ergonomic preference, either hind limb can be used for blood collection. RESULTS: Popliteal sinus venipuncture in anurans can be safely and easily performed with only manual restraint. CLINICAL RELEVANCE: Commonly utilized anuran blood collection techniques include venipuncture of the ventral abdominal vein and lingual venous plexus and cardiocentesis. These techniques can be technically difficult and may pose risks to the patient, particularly if attempted without chemical restraint. Popliteal sinus venipuncture is a reliable, technically simple alternative blood collection technique that can be safely performed in anurans with only manual restraint.

Anesthetic efficacy of dexmedetomidine-ketamine in eastern box turtles (Terrapene carolina carolina) is enhanced with the addition of midazolam and when administered in the forelimb versus the hindlimb.

OBJECTIVE: To compare dexmedetomidine-ketamine (DK; 0.1 and 10 mg/kg, respectively) with midazolam (M; 1.0 mg/kg) or 0.9% sodium chloride (S; 0.2 mL/kg) administered IM in the forelimb (F) or hindlimb (H) in eastern box turtles (Terrapene carolina carolina). ANIMALS: 20 clinically healthy, captive adult eastern box turtles. METHODS: In a randomized, blinded, complete crossover study with 1-week washout periods, turtles were administered each of 3 treatments: F-DKS, F-DKM, or H-DKM. Palpebral reflex, muscle tone, and withdrawal responses were serially assessed and used to calculate cumulative sedation scores at each 5-minute time point. The ability to intubate was evaluated. At 60 minutes, atipamezole (1.0 mg/kg) and either flumazenil (F-DKM, H-DKM; 0.05 mg/kg) or 0.9% sodium chloride (F-DKS; 0.5 mL/kg) were administered IM. RESULTS: All treatments resulted in clinically relevant anesthetic effects. F-DKM produced significantly higher sedation scores than H-DKM or F-DKS at all time points between 10 and 60 minutes (P < .05). Sedation score variability was observed with all treatments with significantly higher variability for H-DKM (P < .05). Intubation was successful in 32, 89, and 11% of turtles in F-DKS, F-DKM, and H-DKM, respectively. Median (range) recovery time was 10 (5-22), 16 (7-45), and 12 (4-28) minutes for F-DKS, F-DKM, and H-DKM, respectively. CLINICAL RELEVANCE: In eastern box turtles, forelimb dexmedetomidine-ketamine resulted in clinically relevant anesthetic effects that were heightened with the addition of midazolam. Hindlimb administration of midazolam-dexmedetomidine-ketamine resulted in reduced and more variable anesthetic effects compared to forelimb administration, supporting a hepatic first-pass effect.

Investigation of Potassium Chloride for Euthanasia of Anesthetized African Clawed Frogs (Xenopus laevis).

Euthanasia is frequently performed in amphibians, but techniques are currently limited in number and variable in effectiveness. The current study examined the use of potassium chloride (KCl) for euthanasia of anesthetized African clawed frogs (Xenopus laevis). Twenty adult, female African clawed frogs were anesthetized by immersion in buffered tricaine methanesulfonate (MS-222) for 5 min beyond loss of righting reflex. Frogs were then randomly assigned to receive one of 4 treatments: KCl via intracardiac injection (10 mEq/kg; IC, n = 5), intracoelomic injection (100 mEq/kg; ICe, n = 5), immersion (4,500 mEq/L; IMS, n = 5), or no treatment (C, n = 5). After treatment, serial heart rate was measured via Doppler device until either the loss of Doppler sounds, a 60-min endpoint (IC, ICe, IMS), or recovery (C). Times to loss of righting reflex, loss of Doppler sounds, and/or recovery were recorded. Plasma potassium concentrations were measured immediately after Doppler sound cessation in frogs in IC (n = 1), ICe (n = 2), and IMS (n = 5). Injection failure occurred in 1 IC frog, and 1 Ice frog regained spontaneous movement 4 min after treatment administration. Data from these 2 frogs were not included in statistical analysis. Doppler sound cessation occurred in 4 of 4, 4 of 4, 0 of 5, and 0 of 5 frogs in IC, ICe, IMS, and C, respectively. Median (range) times to Doppler sound cessation in IC and ICe were 6 (0 to 16) s and 18 (10 to 25) min, respectively. Plasma potassium concentration was greater than 9.0 mmol/L in sampled frogs. Intracardiac KCl at 10 mEq/kg and intracoelomic KCl at 100 mEq/kg were effective for euthanasia of anesthetized African clawed frogs. Return to MS-222 solution after KCl administration may be warranted to prevent unintended, premature anesthetic recovery prior to death.

Characterization of Current Husbandry and Veterinary Care Practices of the Giant Pacific Octopus (Enteroctopus dofleini) Using an Online Survey.

Giant Pacific octopuses (Enteroctopus dofleini) (GPOs) are commonly housed in zoos or aquaria, and sedation, anesthesia, and/or euthanasia may be indicated for a variety of reasons. Despite this need, evidence-based data on best practices is limited and focuses on smaller or more tropical species. The objectives of this study were to survey the aquatic community regarding the husbandry and veterinary care of GPOs, with a specific focus on anesthetic and euthanasia protocols. A two-part web-based survey was distributed to four aquatic and/or veterinary email listservs. Individuals from fifty-two institutions participated in phase one. Results documented that 40 (78 percent) participating institutions currently house GPOs, with most housing one and nine institutions housing two to three GPOs. The median (range) habitat volume is 5405 (1893-16,465) L, and 78 percent of systems are closed. Of the institutions surveyed, 23 have anesthetized or sedated a GPO for nonterminal procedures, including wound care, biopsies, and hemolymph collection. Reported methods of sedation or anesthesia include magnesium chloride, ethanol, isoflurane, tricaine methanesulfonate (MS-222), magnesium sulfate, benzocaine, and dexmedetomidine. Drugs or methods used for euthanasia include magnesium chloride, ethanol, mechanical decerebration, pentobarbital, isoflurane, MS-222, magnesium sulfate, benzocaine, potassium chloride, dexmedetomidine, and freezing. Reported observed side effects include ineffectiveness or inadequate sedation, inking, prolonged drug effects, and behavior changes. Survey data have the potential to guide the husbandry and veterinary care of GPOs and build the framework for future prospective studies on GPO sedation and anesthesia.