Larry J. Minter

The Effects of Feeding on Hematological and Plasma Biochemical Profiles in Green (Chelonia mydas) and Kemp's Ridley (Lepidochelys kempii) Sea Turtles.

In mammals, lipemic blood from sampling too soon after an animal feeds can have substantial effects on biochemical values. Plasma biochemical values in reptiles may be affected by species, age, season, and nutritional state. However, fasting status is not routinely considered when sampling reptile blood. In this paper, we evaluated 2-hour postprandial blood collection in two sea turtle species to investigate the effects of feeding on hematological and plasma biochemical values. Feeding had no significant effects on hematological values in either species, nor did it have an effect on plasma biochemistry values in Kemps ridley sea turtles. In postprandial green turtles, total protein, albumin, ALP, AST, ALT, amylase, and cholesterol increased significantly, and chloride decreased significantly. Although statistically significant changes were observed, the median percent differences between pre- and postprandial values did not exceed 10% for any of these analytes and would not likely alter the clinical interpretation.

The efficacy of alfaxalone for immersion anesthesia in koi carp (Cyprinus carpio)

OBJECTIVE To characterize the physiologic and behavioral effects of a single induction dose and two maintenance doses of alfaxalone delivered by water immersion in the anesthesia of koi (Cyprinus carpio). STUDY DESIGN Prospective, within-subject complete crossover design. ANIMALS Six adult koi (Cyprinus carpio) with a median body weight of 344.5 g (range 292.0-405.0 g). METHODS Koi were immersed in water containing 10 mg L(-1) alfaxalone until immobile and then maintained with alfaxalone at either 1 or 2.5 mg L(-1) via a recirculating water system. Times for anesthetic induction and recovery periods were recorded. Physiologic and blood gas parameters were evaluated before, during and after the anesthetic trial. Response to noxious stimuli was also assessed. RESULTS Median anesthesia induction time for all fish was 5.4 minutes. Median recovery time was 11.8 and 26.4 minutes in the 1.0 and 2.5 mg L(-1) doses, respectively, which were significantly different (p = 0.04). Cessation of opercular movement occurred in 0/6 and 4/6 fish exposed to 1.0 and 2.5 mg L(-1) dose respectively. No difference was observed in median heart rate over the duration of the anesthetic events. Response to noxious stimulation was 4/6 and 0/6 in the 1.0 and 2.5 mg L(-1) doses respectively. Oxygenation and ventilation did not change during the experiment, but there was a significant decrease in blood pH along with an increase in blood lactate concentration. CONCLUSION AND CLINICAL RELEVANCE Administration of alfaxalone, via water immersion, as an induction and maintenance anesthesia agent provided rapid and reliable anesthesia of koi with no mortality. The maintenance dose of 2.5 mg L(-1) was sufficient to prevent response to noxious stimuli but was associated with a clinically relevant depression in opercular rate.

Cystoscopic guided removal of ectopic eggs from the urinary bladder of the Florida cooter turtle (Pseudemys floridana floridana).

Abstract Three gravid, female, wild Florida cooter turtles (Pseudemys floridana floridana) were evaluated and treated by the North Carolina State University College of Veterinary Medicine Turtle Rescue Team as a result of traumatic injuries or infection. As part of medical management, oviposition was induced using oxytocin, which was only partially effective. In all three cases, ectopic eggs were subsequently identified in the urinary bladder by ultrasound and were successfully removed via a minimally invasive cystoscopic-guided technique. One of the three turtles died within several days of the procedure, and necropsy revealed granulomatous bacterial cystitis. It is hypothesized that these complications were likely due to the length of time between induction with oxytocin and the identification and removal of the ectopic egg.

Alfaxalone as an intramuscular injectable anesthetic in koi carp (Cyprinus carpio).

Abstract:  Fish are commonly anesthetized with MS-222 (tricaine methanesulfonate), a sodium-channel-blocker used as an immersion anesthetic, but its mechanism of action as a general anesthetic is uncertain. Alfaxalone is a neurosteroid that acts at the GABAA receptors. Alfaxalone has been evaluated and was deemed successful as an immersion agent in koi carp. Alfaxalone is an effective intramuscular anesthetic in multiple species. A reliable intramuscular anesthetic in fish would be useful in multiple settings. The purpose of this study was to investigate alfaxalone as an intramuscular injectable anesthetic agent in koi carp (Cyprinus carpio). Eight koi carp were utilized in a crossover design. In each trial, six fish received 1 mg/kg, 5 mg/kg, or 10 mg/kg of alfaxalone intramuscularly. They were assessed every 15 min for opercular rate and sedation score. The sedation score was based on a visual scale from 0 to 5, 0 indicating no response and 5 indicating absent righting reflex and anesthesia. Anesthetized koi were placed on a fish anesthesia delivery system (FADS). Time to anesthesia/recovery was recorded and heart rate was recorded every 15 min. Anesthesia was achieved in 0/6, 1/6, and 5/6 fish at 1, 5, and 10 mg/kg, respectively. Duration of anesthesia for one fish at 5 mg/kg was 2 hr. At 10 mg/kg, median anesthesia duration was 6.5 (3–10) hr. At 10 mg/kg, prolonged apnea (2–3 hr) was observed in 3/6 fish, 2/3 died under anesthesia, and 1/3 recovered 10 hr post-injection. Median peak sedation scores were 1.5, 2.5, and 5, at 1, 5, and 10 mg/kg, respectively. A dosage of 10 mg/kg alfaxalone resulted in 33% mortality. The duration of anesthesia and opercular rate were unpredictable. Due to variation in response despite consistent conditions, as well as risk of mortality, intramuscular alfaxalone cannot be recommended for anesthesia in koi carp.

EFFICACY OF ALFAXALONE FOR INTRAVASCULAR ANESTHESIA AND EUTHANASIA IN BLUE CRABS (CALLINECTES SAPIDUS)

The objective of this study was to characterize the behavioral effects and changes in heart rate of four doses of alfaxalone delivered by intravascular injection to blue crabs (Callinectes sapidus). Thirty (male, n = 27; female, n = 3) blue crabs were randomly assigned to one of four treatment groups of alfaxalone: eight animals were assigned to each of the 5-, 10-, and 15-mg/kg treatment groups, and the remaining six animals were assigned to the 100-mg/kg group. Times for anesthetic induction and recovery periods were recorded. Righting reflex, defensive posturing, and heart rate were evaluated before, during, and after the anesthetic trial. Anesthesia was induced in all 14 animals consolidated into the high-dosage group (15 mg/kg [n = 8] and 100 mg/kg [n = 6]), which was significantly greater than 8 of 16 animals in the low-dosage group (5 mg/kg [n = 2] and 10 mg/kg [n = 6]). Median anesthesia induction time for all crabs was 0.4 min, with no significant difference in induction time between groups observed. Median recovery time was 9.4 min (n = 2), 6.1 min (n = 5), 11.3 min (n = 8), and 66.1 min (n = 5) for the 5-, 10-, 15-, and 100-mg/kg groups, respectively. Recovery times were significantly longer for crabs exposed to an induction dose of 100 mg/kg compared with the 10- and 15-mg/kg induction doses. A significant decrease in the median heart rate was observed between the baseline value and that observed at both induction and 5 min postinjection in the 100-mg/kg dose trial. Two mortalities were observed during the anesthesia trials (n = 1, 10 mg/kg; n = 1, 100 mg/kg), both associated with the autotomization of limbs. In summary, the intravascular administration of alfaxalone at 15 mg/kg provided rapid and reliable sedation, whereas alfaxalone administered at 100 mg/kg produced rapid and long lasting anesthesia.

Effects of Intramuscular Meloxicam Administration on Prostaglandin E2 Synthesis in the North American Bullfrog (Rana catesbeiana)

Abstract:  Meloxicam is a commonly used nonsteroidal anti-inflammatory drug (NSAID) in veterinary medicine, but its use in amphibians has not been reported in the literature. NSAIDs are known to act by providing anti-inflammatory and analgesic actions by inhibiting the synthesis of prostaglandin E2 (PGE2). The objective of this study was to evaluate whether the intramuscular administration of meloxicam would decrease the circulating serum PGE2 levels in the North American bullfrog (Rana catesbeiana) following tissue trauma induced by a punch biopsy. Eighteen adult North American bullfrogs were randomly assigned to two treatment groups: meloxicam (0.1 mg/kg i.m.) and control (0.9% saline i.m.). Blood was obtained via cardiocentesis immediately prior to administration of the two treatment regimes and serum was frozen. A 4-mm punch biopsy was taken from the right triceps femoris muscle to induce an inflammatory response. Twenty-four hours later, a second blood sample was collected and serum was harvested and frozen. Serum PGE2 concentrations were measured using a commercial PGE2 enzyme assay (EIA) kit. Twenty-four hours following the biopsy, the mean circulating PGE2 levels of animals treated with meloxicam was 57.79 ± 12.35 pg/ml, which did not differ significantly from animals that were treated with saline (85.63 ± 17.55 pg/ml, P ≥ 0.05). The calculated means of the absolute change between the circulating baseline PGE2 levels and the postinjury circulating PGE2 levels were significantly lower in animals treated with meloxicam (13.11 ± 17.31 pg/ml) than in control animals treated with saline (46.14 ± 38.02 pg/ml) (P ≤ 0.05). These results suggest that the systemic administration of meloxicam at a dosage of 0.1 mg/kg once daily suppresses circulating serum PGE2 levels postinjury in the North American bullfrog.

Anaerobic oral flora in the North American black bear (Ursus americanus) in eastern North Carolina.

Microbial flora can provide insight into the ecology and natural history of wildlife in addition to improving understanding of health risks. This study examines the anaerobic oral flora of hunter killed black bears (Ursus americanus) in eastern North Carolina. Oral swabs from the buccal and lingual supragingival tooth surfaces of the first and second mandibular and maxillary molars of 22 black bears were inoculated onto Brucella Blood Agar plates supplemented with hemin and vitamin K after transport from the field using reduced oxoid nutrient broth. Sixteen anaerobic bacterial species, representing nine genera were identified using the RapID ANA II Micromethod Kit system and a number of organisms grown that could not be identified with the system. The most frequently identified anaerobes were Peptostreptococcus prevotii, Streptococcus constellatus, and Porphyromonas gingivalis. The diversity in the anaerobic oral flora of black bear in eastern North Carolina suggests the importance of including these organisms in basic health risk assessment protocols and suggests a potential tool for assessment of bear/habitat interactions.

Surgical Repair of a Cleft Palate in an American Bison (Bison bison)

Abstract A 4-mo-old bison (Bison bison) was evaluated and treated at a university veterinary hospital for a cleft defect in the hard and soft palate. Using a mandibular symphysiotomy approach, the palatal defect was repaired with a Z-plasty pattern in the soft palate and mucoperiosteal flaps in the hard palate. A small area of dehiscence in the rostral aspect of the hard palate, and aspiration pneumonia, were complications, but the bison calf recovered with medical management. Even though this surgical procedure has a high potential for complications, the described technique allowed return to normal feeding and resolution of the aspiration pneumonia by 14 mo postsurgery.

CYSTIC UROLITHIASIS IN CAPTIVE WAXY MONKEY FROGS (PHYLLOMEDUSA SAUVAGII)

Abstract:  The waxy monkey frog (Phyllomedusa sauvagii) is an arboreal amphibian native to arid regions of South America, and it has developed behavioral and physiologic adaptations to permit survival in dry environments. These adaptations include a uricotelic nitrogen metabolism and unique cutaneous lipid excretions to prevent evaporative water loss. Uroliths are a rare finding in amphibians. Six adult, presumed wild-caught waxy monkey frogs housed in a museum animal collection were diagnosed with cystic urolithiasis over a 7-yr period, and a single animal was diagnosed with four recurrent cases. Six cases were identified incidentally at routine physical or postmortem examination and four cases were identified during veterinary evaluation for coelomic distension, lethargy, anorexia, and increased soaking behavior. Calculi were surgically removed from three frogs via cystotomy, and a single frog underwent three cystotomies and two cloacotomies for recurrent urolithiasis. Two frogs died within the 24-hr postoperative period. Two representative calculi from a single frog were submitted for component analysis and found to consist of 100% ammonium urate. In the present report, cystic calculi are proposed to be the result of a high-protein diet based on a single invertebrate source, coupled with uricotelism, dehydration, increased cutaneous water loss, body temperature fluctuations facilitating supersaturation of urine, and subsequent accumulation and precipitation of urogenous wastes within the urinary bladder. Surgical cystotomy represents a short-term treatment strategy for this condition. Preventative measures, such as supplying a diversified and balanced diet in addition to environmental manipulation aimed at promoting adequate hydration, are anticipated to be more-rewarding management tools for cystic urolithiasis in the waxy monkey frog.

Collection and characterization of semen from Chilean rose tarantulas (Grammostola rosea)

OBJECTIVE To establish a nonterminal semen collection method for use in captive Chilean rose tarantulas (Grammostola rosea) and to evaluate tools for investigating morphology and viability of spermatozoa. ANIMALS 7 mature male Chilean rose tarantulas. PROCEDURES Each tarantula was anesthetized in a 500-mL induction chamber containing a cotton ball infused with 2 mL of isoflurane. Semen collection was performed by applying direct pressure to the palpal bulbs (sperm storage organs) located on the distal segment of the palpal limbs. Morphology of spermatozoa was examined by light microscopy and transmission and scanning electron microscopy. Propidium iodide and a fluorescent membrane-permeant nucleic acid dye were used to evaluate cell viability. RESULTS Semen was collected successfully from all 7 tarantulas. Microscopic examination of semen samples revealed coenospermia (spherical capsules [mean ± SD diameter, 10.3 ± 1.6 μm] containing many nonmotile sperm cells [mean number of sperm cells/capsule, 18.5 ± 3.8]). Individual spermatozoa were characterized by a spiral-shaped cell body (mean length, 16.7 ± 1.4 μm; mean anterior diameter, 1.5 ± 0.14 μm). Each spermatozoon had no apparent flagellar structure. The fluorescent stains identified some viable sperm cells in the semen samples. CONCLUSIONS AND CLINICAL RELEVANCE The described technique allowed simple and repeatable collection of semen from Chilean rose tarantulas. Semen from this species was characterized by numerous spherical capsules containing many nonmotile spermatozoa in an apparently quiescent state. Fluorescent staining to distinguish live from dead spermatozoa appeared to be a useful tool for semen evaluation in this species.