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.

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.

Comparative nutrient analysis of four species of cockroaches used as food for insectivores by life stage, species, and sex.

A variety of insects are fed to insectivorous animals; however, nutritional analyses are often limited to adult life stages. Four species of nymph and adult female and male cockroaches (Blaberus giganteus, Blaptica dubia, Blatta lateralis, and Gromphadorhina portentosa) were analyzed for moisture, crude protein (CP), acid detergent fiber (ADF), fat, ash, and mineral content. The small sample size of this study precluded statistical analyses, however comparatively, CP in adult B. lateralis and B. dubia was greater than the CP in nymphs of the same species. Adult and nymph B. dubia had the greatest CP (96.6% and 65.3%, respectively) compared to the other three species. Inversely, fat content in adult B. lateralis and B. dubia was lower than that of nymphs of the same species. All adults contained similar levels of ADF, yet adult B. giganteus had greater ADF than nymphs; conversely, B. lateralis and B. dubia adults had less ADF than nymphs. There were differences noted in mineral parameters among the four species and life stages. Adult B. giganteus had less Ca than G. portentosa, and the lowest Ca content of the four species of nymphs and adults analyzed. This study underscores the differences in nutrient content with respect to life stage and species in previously understudied cockroaches to improve nutrient intake in captive insectivores.

Cane toad (Rhinella marina) vitamin A, vitamin E, and carotenoid kinetics.

Many amphibian species are threatened with extinction. Understanding their vitamin A (retinol), E (alpha-tocopherol), and carotenoid requirements is vital, as normal levels of these nutrients have a known connection to breeding success with abnormal levels leading to disease. This research examined vitamins A, E, and carotenoids (apocarotenoid, beta-carotene; beta-cryptoxanthin, lutein, zeaxanthin, and esters) concentration kinetics in the liver and plasma of 65 (57.8) cane toads (Rhinella marina) over 4 months supplemented with commercially available invertebrates in human care. Cane toads were opportunistically collected as part of a population control program for use as an amphibian model species. Toads were randomly assigned to one of two diets: treatment 1 was brown house crickets (Acheta domesticus) consuming Mazuri® Hi Calcium Gut Loading Diet without vitamin A or E supplement, plus fresh raw vegetables (carrot/sweet potato); Treatment 2 was the same diet except no vegetables. Ten toads were euthanized on Day 0 to analyze baseline free-ranging liver and plasma metabolites. Six toads consuming each treatment were euthanized on Days 22, 50, and 81, and n = 7 on Day 119 for analysis. Regardless of dietary treatment, most liver and blood metabolites were substantially higher at time 0 than all time points thereafter (p < .05); Ex: liver vitamin A at time 0 was 87.7 ± 16.12 µg/g while Day 119 for treatments 1 and 2 were 11.6 ± 1.19 and 8.2 ± 0.74, respectively. Few statistically significant differences between diets at the same time point were noted (p < .05). The results from this study indicate that additional or alternative diet supplementation may be needed for cane toads (and potentially other amphibians) to mimic their free-ranging diets.

Retrospective Analysis of Cataract Formation and Nutritional Etiology in a Managed Collection of Parakeet Auklets (Aethia psittacula).

The parakeet auklet (Aethia psittacula) is a piscivorous seabird with a natural diet of various invertebrate and teleost species, which is challenging to replicate in a managed collection. A high prevalence of early onset cataracts was observed in a managed collection of parakeet auklets at the North Carolina Zoo (Asheboro, NC, USA), which was hypothesized to be related to inappropriate vitamin A and E levels. From 1994 to 2002, these parakeet auklets were offered dietary supplementation comprising Vita-Zu small bird tablets. In June 2002, the birds were transitioned to only Thiamin-E paste (vitamin E and thiamin only). Plasma samples were collected from birds with and without cataracts from 1998 to 2005 and submitted for vitamin A (retinol) and vitamin E (α-tocopherol) analysis. Food items comprising the birds' diet were also evaluated for vitamin content. This information was combined with clinical and necropsy data from medical records from 1994 to 2015. A total of 78% of birds (39/50) developed cataracts, with a median age of onset of 7 years (range, 2-12 years). Cataracts ranged from incipient to hypermature during both routine ophthalmic examinations and postmortem evaluations. The median (range) of plasma retinol and α-tocopherol values were 1.99 µg/mL (0.20-6.68 µg/mL) and 15.39 µg/mL (3.40-96.27 µg/mL), respectively. There were no significant differences in plasma concentrations of vitamins based on the animals' sex, origin, presence of cataracts, or administered vitamin supplementation product. No other etiologies for cataract development were identified in the population. Further research in free-ranging parakeet auklet nutrition and cataract occurrence is warranted for continued species collection management.

BLOOD FATTY ACID PROFILES OF NERITIC JUVENILE WILD GREEN TURTLES (CHELONIA MYDAS) AND KEMP'S RIDLEYS (LEPIDOCHELYS KEMPII).

Blood fatty acid profiles can indicate an animal's wild-type diet composition and fatty acid status, but have not been reported in sea turtles. Newer technologies allow for fatty acid profiles from very small (less than three drops) samples of whole blood. This study examined whole blood fatty acid profiles of presumably healthy, neritic, juvenile, wild green (Chelonia mydas) (n = 9; 6 males, 3 females) and Kemp's ridley (Lepidochelys kempii) (n = 8; 6 males, 2 females) turtles from North Carolina, USA. Saturated fatty acids, which can be synthesized de novo, consisted primarily of 16:0, although green turtle blood had a higher proportion of 18:0 (P < 0.001) than Kemp's ridleys, while Kemp's ridley blood had higher proportions of 17:0 (P = 0.007), 20:0 (P = 0.03), 22:0 (P= 0.002), and 24:0 (P < 0.001) as compared with green turtles. Total monounsaturated w7 fatty acids, which can be synthesized de novo or may be diet derived, were higher in Kemp's ridleys and predominantly in the form of 16:1 and 18:1w7 fatty acids. Kemp's ridley blood had more than double the relative proportion of 16:1w7 as compared with green turtles (P= 0.03). Green turtles had higher levels of 18:2w6 than Kemp's ridleys (P= 0.02). In both turtle species, 20:4w6 was detected, despite predicted low dietary proportions, suggesting bioconversion from precursors. Finally, green turtles had higher levels of 18:3w3 compared with Kemp's ridleys, while Kemp's ridleys had higher proportions of 20:5w3 compared with green turtles (P < 0.001, = 0.007, respectively). Whole blood fatty acid profiles generally correlate to previous work with lipid depots, supporting the use of this less invasive methodology to advance the understanding of fatty acid nutrition of sea turtles. These data can be used to assess and guide nutrition and health programs for sea turtles under human care.

URINARY CORTICOSTERONE CONCENTRATIONS IN FREE-RANGING AND MANAGED CANE TOADS (RHINELLA MARINA).

Approximately 40% of amphibian species are threatened with extinction. The welfare of amphibians maintained under managed care as assurance populations is of vital importance to guard against extinction and provide a viable source population for future reintroduction. To manage amphibian species properly ex situ, it is important to understand how stress levels change over time when animals are removed from the wild and placed into managed environments. Corticosterone was analyzed in urine samples from free-ranging cane toads (Rhinella marina, n = 55) in Miami, FL, and under managed care for 22 (n = 48), 50 (n = 11), 81 (n = 25), or 119 (n = 10) days. Concentrations of corticosterone in free-ranging toad urine averaged 1.74 ± 0.195 ng/ml urine specific gravity (sp. gr.), which was greater (P < 0.05) than other time points (day 22: 0.77 ± 0.114 L; day 81: 0.85 ± 0.191 ng/ml sp. gr.; day 119: 0.58 ± 0.093 ng/ml sp. gr.), except day 50 (0.91 ± 0.274 ng/ml sp. gr.), which was not different from free-ranging or managed care values. Thus, corticosterone was lower in cane toads under managed care compared with those sampled in the wild, suggesting that managed care is not a stressor for this species.

Circulating nutrients and hematological parameters in managed African elephants (Loxodonta Africana) over a 1-year period.

African elephants (Loxodonta Africana) are currently considered a vulnerable species. One key to improving methods of species management is to better monitor and understand elephant nutrition. Analyzing circulating nutrients is one of the best and least invasive methods of monitoring managed elephant nutrition, but limited reference values are available. This study examined the circulating basic hematology concentrations, minerals, vitamins A, D, and E, and fatty acids of six African elephants (two males and four females) at the North Carolina Zoo collected monthly from March 2016 to April 2017 and compared levels among seasons. Creatinine (CRE) and albumin had seasonal differences (p ≤ .05). Calcium, magnesium, phosphorus, potassium, selenium, zinc, cobalt, manganese, and molybdenum displayed seasonal differences (p ≤ .05). Retinol and 25-hydroxyvitamin D2 had seasonal differences (p ≤ .05). Linoleic acid, α-linolenic acid, arachidonic acid, total omega-3 fatty acids, total omega-6 fatty acids, and the omega-6 to omega-3 ratio showed seasonal differences (p ≤ .05). Findings suggest that exogenous vitamin E supplements may not be necessary with a mixed feedstuff diet (hay, fortified concentrate pellet, browse, and produce) based on circulating values. This data offer updated information on circulating reference values and novel circulating concentrations of nutrients for Southeastern US managed African elephants that can be used to inform nutritional and health management in all similar habitats.

Analyses of African elephant (Loxodonta africana) diet with various browse and pellet inclusion levels.

To more closely simulate the diet of free-ranging elephants, the diet of six (2.4) African elephants (Loxodonta africana) was altered to include more browse and less pelleted complete feed (5% total diet). Dietary proximate compounds, minerals, vitamins A (and carotenoids), D and E, and fatty acids were analyzed on pelleted diet items and forages including hay, grass, and browse. A total of 42 browse species were offered over 1 year with an average total diet inclusion of 5.2% (dry matter basis) per day. Dietary Na and Se were low while Fe and Mn were high compared to published intake levels for elephants. Analyzed nutrients within browse varied widely among seasons and species. Ingredient analyses were used to create predicted elephant nutrient intake for (a) the current diet, (b) a diet excluding pellets, and (c) a diet excluding pellets and providing browse at doubled levels. Formulated diets excluding pellets had lower mineral levels than the current diet and doubled browse did not alter mineral inclusions of concern. This study provides seasonal data on the nutrient levels of Southeastern browse species important for various pachyderm and herbivorous species. Predicted nutrient intake with new diet scenarios does not support the exclusion of pellets in the diets of African elephants without greater browse quantity availability, strict diet management, or additional supplements.

ANALYSIS OF FATTY ACID PROFILES FOR EASTERN BOX (TERRAPENE CAROLINA CAROLINA) AND COMMON SNAPPING (CHELYDRA SERPENTINE) TURTLES IN WILD AND MANAGED CARE ENVIRONMENTS.

The housing of wild animals in managed care facilities requires attention to all aspects of husbandry. Diets of wild animals often differ in composition, consistency, and quantity when compared with those in managed care settings including zoos, rehabilitation facilities, and aquaria. It was hypothesized that dietary differences from wild versus managed care would be reflected in data of circulating fatty acids based on previous studies. The current study examined the effect of species and environment on fatty acid concentrations in two omnivorous species of chelonians: Eastern box turtles, Terrapene carolina carolina, and common snapping turtles, Chelydra serpentina, located in the wild and managed care. Whole blood was collected and placed on spot cards for analysis of 26 fatty acids in a total lipid fatty acid profile. The present research indicated that Eastern box turtles have significantly (P < 0.05) higher percentages of linoleic acid (18:2n6), eicosadienoic acid (20:2n6), and mead acid (20:3n9). Common snapping turtles have significantly (P < 0.05) higher percentages of myristic acid (14:0), dihomo-γ-linolenic acid (20:3n6), erucic acid (22:1n9), and n-6 docosapentaenoic acid (22:5n6). Environmental effects also were noted; wild turtles had higher percentages of α-linolenic acid (18:3n3), arachidic acid (20:0), eicosadienoic acid (20:2n6), and eicosatrienoic acid (20:3n3) (P < 0.05), whereas n-6 docosapentaenoic acid (22:5n6) was higher for the managed care group. Eicosadienoic acid (20:2n6), behenic acid; 22:0), adrenic acid (22:4n6), n-6 docosapentaenoic acid (22:5n6), and nervonic acid (24:1) were significantly different (P < 0.05) in species-environment interactions without any noted species or environment patterns. Fatty acids are useful for many important biological functions including proper immune system regulation, and therefore, the present research provides medically relevant data for reptile diagnostics. This research may help further improve diets of all chelonians kept in managed care, regardless of species.

Cystic urolithiasis in captive waxy monkey frogs (Phyllomedusa sauvagii).

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.

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.

Identifying and managing an adverse food reaction in a polar bear (Ursus maritimus) by an elimination diet trial.

A 16-yr-old polar bear (Ursus maritimus) presented with severe diarrhea shortly following transfer to the North Carolina Zoological Park. Multiple diagnostic procedures were performed over several months and the cause of the chronic diarrhea was inconclusive. Histologically, colonic mucosal biopsies were consistent with severe chronic eosinophilic and lymphoplasmacytic colitis with no evidence of etiologic agents present. A dietary elimination trial was conducted and an adverse food reaction to the dog chow in the diet was confirmed.

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

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 GABA(A) 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 10mg/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.

Performance of a manually operated salad spinner centrifuge for serum separation in the healthy domestic horse (Equus caballus) and southern white rhinoceros (Ceratotherium simum).

BACKGROUND: Field veterinarians and researchers studying wild species, such as the southern white rhinoceros, often work in remote areas with limited access to standard laboratory equipment, hindering the ability to measure serum analytes. OBJECTIVES: The first objective was to produce an inexpensive, manually operated centrifuge that could accept standard laboratory tubes by modifying a consumer-grade salad spinner with low-cost materials. The second objective was to compare biochemistry analysis results obtained from equine and southern white rhinoceros serum separated by traditional laboratory and manual salad spinner centrifugation. METHODS: We optimized the design and serum separation protocol using non-anticoagulated equine blood. Equine and rhinoceros serum samples were separated by manual salad spinner or traditional laboratory centrifugation. Measured analytes included sodium, potassium, chloride, urea nitrogen, creatinine, phosphorous, total calcium, magnesium, glucose, total protein, albumin, globulin, creatinine kinase, aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, gamma-glutamyl transferase, total bilirubin, bicarbonate, sorbitol dehydrogenase, and triglycerides. Results obtained from serum separated by each centrifugation technique were compared by Deming regression and Bland-Altman analyses. RESULTS: A tube adaptor insert modeled after a swing angle rotor and a two-step salad spinner centrifugation yielded serum comparable to traditional laboratory centrifugation. For the majority of analytes, no proportional or constant biases were detected between centrifugation methods. A positive proportional bias in the measurement of ALP in serum separated by manual centrifugation was detected in both equine and rhinoceros samples. CONCLUSIONS: Manual centrifugation with a modified salad spinner yields diagnostic quality serum suitable for the measurement of most standard biochemistry analytes.

Effects of age, seasonality, and reproductive status on the gut microbiome of Southern White Rhinoceros (Ceratotherium simum simum) at the North Carolina zoo.

BACKGROUND: Managed southern white rhinoceros (Ceratotherium simum simum) serve as assurance populations for wild conspecifics threatened by poaching and other anthropocentric effects, though many managed populations experience subfertility and reproductive failure. Gut microbiome and host health are inextricably linked, and reproductive outcomes in managed southern white rhinoceros may be mediated in part by their diet and gut microbial diversity. Thus, understanding microbial dynamics within managed populations may help improve conservation efforts. We characterized the taxonomic composition of the gut microbiome in the managed population of female southern white rhinoceros (n = 8) at the North Carolina Zoo and investigated the effects of seasonality (summer vs. winter) and age classes (juveniles (n = 2; 0-2 years), subadults (n = 2; 3-7 years), and adults (n = 4; >7 years)) on microbial richness and community structure. Collection of a fecal sample was attempted for each individual once per month from July-September 2020 and January-March 2021 resulting in a total of 41 samples analyzed. Microbial DNA was extracted and sequenced using the V3-V4 region of the 16S rRNA bacterial gene. Total operational taxonomic units (OTUs), alpha diversity (species richness, Shannon diversity), and beta diversity (Bray-Curtis dissimilarity, linear discriminant analysis effect size) indices were examined, and differentially enriched taxa were identified. RESULTS: There were differences (p < 0.05) in alpha and beta diversity indices across individuals, age groups, and sampling months. Subadult females had higher levels of Shannon diversity (Wilcoxon, p < 0.05) compared to adult females and harbored a community cluster distinct from both juveniles and adults. Samples collected during winter months (January-March 2021) possessed higher species richness and statistically distinct communities compared to summer months (July-September 2020) (PERMANOVA, p < 0.05). Reproductively active (n = 2) and currently nonreproductive adult females (n = 2) harbored differentially enriched taxa, with the gut microbiome of nonreproductive females significantly enriched (p = 0.001) in unclassified members of Mobiluncus, a genus which possesses species associated with poor reproductive outcomes in other animal species when identified in the cervicovaginal microbiome. CONCLUSION: Together, our results increase the understanding of age and season related microbial variation in southern white rhinoceros at the North Carolina Zoo and have identified a potential microbial biomarker for reproductive concern within managed female southern white rhinoceros.

Investigation of euthanasia techniques in four species of cockroaches.

While cockroaches are commonly exhibited in zoos and museums, studied in research laboratories, and even kept as pets, scientifically based guidelines for their euthanasia are lacking. This study assessed euthanasia techniques in four species of cockroaches (Dubia (Blaptica dubia), red runner (Shelfordella lateralis), Madagascar hissing (Gromphadorhina portentosa), and giant cave (Blaberus giganteus)). In an initial pilot study, two hundred fifty adult Dubia cockroaches were exposed in groups of ten to a cotton ball soaked with 2 mL of isoflurane in a 1 L air-tight chamber. Thirty minutes beyond loss of any individual movement, groups were exposed to one of the following secondary treatments: freezing at -18 °C or -80 °C from 0.25 to 24 hours; immersion in 10% neutral buffered formalin, 70% isopropyl alcohol, or reverse osmosis water for 0.25 or 0.5 hours; or intracoelomic injection of potassium chloride (456 mEq/kg) or pentobarbital-based euthanasia solution (3.9 g/kg). A control group remained in the air-tight isoflurane chamber for 24 hours. Following all treatments, cockroaches were monitored for an additional 24 hours for spontaneous movement. Irreversible loss of movement was considered synonymous with irreversible loss of consciousness (death). Across all species, isoflurane anesthesia followed by either 70% isopropyl alcohol immersion for 0.25 or 0.5 hours or isoflurane exposure for 24 hours resulted in euthanasia in 100% of cockroaches. This study is the first evaluation of American Veterinary Medical Association-recommended euthanasia protocols in cockroaches.

Comparison of Corticosterone Concentrations in Dermal Secretions and Urine in Free-Ranging Marine Toads (Rhinella marina) in Human Care.

Corticosterone concentrations have been measured in amphibians by collecting blood or urine samples. However, blood sampling is invasive, and urine can be difficult to collect. A novel method of swabbing the skin of an amphibian has been utilized in numerous species but has not been verified in marine toads (Rhinella marina). This pilot study tested dermal swabs as a noninvasive method for collecting and measuring dermal corticosterone secretions. Swabs were used to collect dermal secretion samples from sixty-six free-ranging marine toads collected on Zoo Miami grounds. The subsequent day the toads were shipped to the North Carolina Zoo where dermal samples were collected again. Additional dermal and urine samples were collected on days 9, 15, 32, and 62 under human care to measure corticosterone concentrations. There was no significant correlation (P ≥ 0.05) noted between corticosterone concentrations reported in dermal swabs and those in urine samples at all four of the euthanasia time points or between the corticosterone concentrations reported in either urine or dermal swabs and the weight of the toads. Dermal swab concentrations (ng/mL) were significantly higher (P ≤ 0.05) on the day of capture (0.64 ± 0.03) and the day of arrival (0.67 ± 0.03) than on day 15 (0.47 ± 0.03). The urine corticosterone concentrations decreased while the toads were in human care with a significant decrease (P ≤ 0.05) between days 9 (0.45 ± 0.07) and 32 (0.21 ± 0.06). This study demonstrated that dermal swabs can be used to collect marine toad corticosterone concentration samples.

Reye's or Reye's-like syndrome in western lowland gorilla (Gorilla gorilla gorilla).

BACKGROUND: A 15-year-old western lowland gorilla (Gorilla gorilla gorilla) died shortly after transfer to the North Carolina Zoo. METHODS: Complete blood count, serum biochemical analysis, and necropsy were performed. RESULTS: Combination of compatible clinical signs, biochemical and histopathological findings fulfilled all of the CDC definition criteria of Reye's or a Reye's like syndrome. CONCLUSIONS: This report describes a case of Reye's syndrome or Reye's-like syndrome in a non-human primate.