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Briefly considered extinct in the wild, the future of the wild population of the Wyoming toad (Anaxyrus baxteri) continues to rely on captive breeding to supplement the wild population. Given its small natural geographic range and history of rapid population decline at least partly due to fungal disease, investigation of the diversity of key receptor families involved in the host immune response represents an important conservation need. Population decline may have reduced immunogenetic diversity sufficiently to increase the vulnerability of the species to infectious diseases. Here we use comparative transcriptomics to examine the diversity of toll-like receptors and major histocompatibility complex (MHC) sequences across three individual Wyoming toads. We find reduced diversity at MHC genes compared to bufonid species with a similar history of bottleneck events. Our data provide a foundation for future studies that seek to evaluate the genetic diversity of Wyoming toads, identify biomarkers for infectious disease outcomes, and guide breeding strategies to increase genomic variability and wild release successes.

Objective To document ocular lesions present in cold-stunned sea turtles and determine the impact of these lesions on their release. Animals All sea turtles (Caretta caretta, Chelonia mydas, and Lepidochelys kempii) presenting to rehabilitation centers in North Carolina over two cold stun seasons. Procedures Complete ophthalmic examination using slit-lamp biomicroscopy, fluorescein stain, rebound tonometry, and occasionally binocular indirect funduscopy was performed within 1 week of presenting to the rehabilitation centers. A second examination was performed 2 weeks after the first examination in animals with ocular lesions and still present at the center. Results One hundred and sixty-four turtles (121 Chelonia mydas, 26 Lepidochelys kempii, 17 Caretta caretta) were evaluated over two cold stun seasons (2016-2017 and 2017-2018). Ocular or periocular lesions were identified in 78 of 164 (47.5%) turtles examined with 37 of 164 (22.5%) having bilateral disease. The most common ocular lesion was superficial corneal ulceration, accounting for 21.3% of all lesions. Adnexal lesions were also common, while intraocular abnormalities were overall rare. Most resolved uneventfully. Though some blinding lesions were noted, these were not present bilaterally in any one turtle, and thus, ocular findings did not affect release. Conclusion Ocular lesions in cold-stunned sea turtles are common. Some of these lesions are potentially blinding or require appropriate care, so evaluation of cold-stunned turtles should include a complete ophthalmic examination.

Sea turtles are frequently presented for rehabilitation with injuries for which analgesic treatment is warranted. Ketoprofen is a nonsteroidal anti-inflammatory drug (NSAID) widely used in clinical veterinary medicine for musculoskeletal pain relief. Pharmacokinetics of 2 mg/kg IM have been studied in loggerhead sea turtles (Caretta caretta) as a single and a repeated dose q24hr for 3 days. Safety of longer term administration has not been performed, however, and NSAID use carries a risk of potential complications, including gastrointestinal ulceration, kidney damage, and bleeding. The objective of the current study was to determine the effects of a 5-day course of ketoprofen on thromboelastography (TEG) and hematological (including thrombocytes) and plasma biochemical analytes in loggerheads. A secondary objective was to determine 24-hr trough concentrations of ketoprofen after 5 days of treatment. Eight loggerheads were treated with ketoprofen 2 mg/kg IM q24hr for 5 days, and TEG, hematology, and plasma biochemistry panels were performed before and at the conclusion of treatment. Eight controls were treated with an equivalent volume of saline intramuscularly. Virtually no changes were detected before and after treatment or between treatment and control groups in any of the 24 endpoints evaluated, and marginal differences were not considered clinically relevant. Plasma ketoprofen concentrations after 5 days of treatment indicated no accumulation over that duration. Ketoprofen at 2 mg/kg IM q24hr for up to 5 days in loggerheads appears safe with respect to blood clotting and blood data, although other potential effects were not evaluated.

Sea turtles are frequently presented for rehabilitation with injuries for which analgesic treatment is warranted. Ketoprofen is a nonsteroidal anti-inflammatory drug (NSAID) widely used in clinical veterinary medicine for musculoskeletal pain relief. Pharmacokinetics of 2 mg/kg IM have been studied in loggerhead sea turtles (

Fibropapillomatosis is associated with chelonid alphaherpesvirus 5 (ChHV5) and tumor formation in sea turtles. We collected blood samples from 113 green (Chelonia mydas) and 112 loggerhead (Caretta caretta) turtles without fibropapillomatosis, including 46 free-ranging turtles (20 green turtles, 26 loggerheads), captured in Core Sound, North Carolina, and 179 turtles (93 green turtles, 86 loggerheads) in rehabilitative care in North Carolina. Blood samples were analyzed for ChHV5 DNA using quantitative polymerase chain reaction (qPCR), and for antibodies to ChHV5 peptides using an enzyme-linked immunosorbent assay (ELISA). None of the samples from foraging turtles tested positive for ChHV5 by qPCR, ELISA was not used for foraging turtles. Samples from 18/179 (10.1%) rehabilitating turtles tested positive for ChHV5 using qPCR, and 32/56 (57.1%) rehabilitating turtles tested positive for antibodies to ChHV5 using ELISA. Five turtles that tested positive by qPCR or ELISA at admission converted to being undetectable during rehabilitation, and five that initially tested negative converted to being positive. Both sea turtle species were significantly more likely to test positive for ChHV5 using ELISA than with qPCR (p &lt, 0.001). There was no difference in the proportions of green turtles versus loggerheads that tested positive for ChHV5 using qPCR, but loggerheads were significantly more likely than green turtles to test positive for ChHV5 using ELISA. This finding suggests that loggerheads infected with ChHV5 at some point in their life may be more able than green turtles to mount an effective immune response against recrudescent infection, pointing to species-specific genetic differences in the two species&rsquo, immune response to ChHV5 infection. This is the first study to analyze antibodies to ChHV5 in loggerhead turtles and represents the most complete dataset on ChHV5 DNA detection in sea turtles encountered in the more northern latitudes of their western Atlantic habitat.

Ketoprofen is a nonsteroidal anti-inflammatory and analgesic agent that nonselectively inhibits cyclooxygenase, with both COX-1 and COX-2 inhibition. Recent studies on COX receptor expression in reptiles suggest that nonselective COX inhibitors may be more appropriate than more selective inhibitors in some reptiles, but few pharmacokinetic studies are available. The goal of this study was to determine single- and multidose (three consecutive days) pharmacokinetics of racemic ketoprofen administered intravenously and intramuscularly at 2 mg/kg in healthy juvenile loggerhead turtles (Caretta caretta). The S-isomer is the predominant isomer in loggerhead sea turtles, similar to most mammals, despite administration of a 50:50 racemic mixture. Multidose ketoprofen administration demonstrated no bioaccumulation; therefore, once-daily dosing will not require dose adjustment over time. S-isomer pharmacokinetic parameters determined in this study were Cmax of 10.1 μg/ml by IM injection, C0 of 13.4 μg/ml by IV injection, AUC of 44.7 or 69.4 μg*hr/ml by IM or IV injection, respectively, and T½ of 2.8 or 3.6 hr by IM or IV injection, respectively. Total ketoprofen plasma concentrations were maintained for at least 12 hr above concentrations determined to be effective for rats and humans. A dose of 2 mg/kg either IM or IV every 24 hr is likely appropriate for loggerhead turtles.

OBJECTIVE To compare physiologic and anesthetic effects of alfaxalone administered IV to yearling loggerhead sea turtles (Caretta caretta) at 3 different doses. DESIGN Randomized crossover study. ANIMALS 9 healthy yearling loggerhead sea turtles. PROCEDURES Animals received each of 3 doses of alfaxalone (3 mg/kg [1.4 mg/lb], 5 mg/kg [2.3 mg/lb], or 10 mg/kg [4.5 mg/lb]) administered IV in randomly assigned order, with a minimum 7-day washout period between doses. Endotracheal intubation was attempted following anesthetic induction, and heart rate, sedation depth, cloacal temperature, and respirations were monitored. Times to first effect, induction, first voluntary muscle movement, first respiration, and recovery were recorded. Venous blood gas analysis was performed at 0 and 30 minutes. Assisted ventilation was performed if apnea persisted 30 minutes following induction. RESULTS Median anesthetic induction time for all 3 doses was 2 minutes. Endotracheal intubation was accomplished in all turtles following induction. Heart rate significantly increased after the 3- and 5-mg/kg doses were administered. Median intervals from alfaxalone administration to first spontaneous respiration were 16, 22, and 54 minutes for the 3-, 5-, and 10-mg/kg doses, respectively, and median intervals to recovery were 28, 46, and 90 minutes, respectively. Assisted ventilation was required for 1 turtle after receiving the 5-mg/kg dose and for 5 turtles after receiving the 10-mg/kg dose. The 10-mg/kg dose resulted in respiratory acidosis and marked hypoxemia at 30 minutes. CONCLUSIONS AND CLINICAL RELEVANCE IV alfaxalone administration to loggerhead sea turtles resulted in a rapid anesthetic induction and dose-dependent duration of sedation. Assisted ventilation is recommended if the 10 mg/kg dose is administered.

Four captive raptors, an American kestrel ( Falco sparverius ), peregrine falcon ( Falco peregrinus ), golden eagle ( Aquila chrysaetos ), and barn owl ( Tyto alba ), were diagnosed with ventricular and/or proventricular foreign material impactions consisting of artificial turf substrate, paper and plastic substrate, grass, and newspaper. Partial or total anorexia was reported in all birds and decreased casting in 2 birds. Survey radiographs confirmed presence of gastric enlargement in all 4 birds. The kestrel and eagle were treated unsuccessfully with gastroscopy and gastric lavage, respectively, followed by surgical intervention to remove the ventricular impactions. Both birds died of undetermined causes after surgery. The peregrine falcon died before medical or surgical intervention was started, and the owl was managed successfully with oral mineral oil and liquid diet to facilitate egestion of the foreign material as a pellet. Lead poisoning was suspected as the predisposing cause for foreign body ingestion in the eagle, but underlying causes for pica in the other birds were not determined. Radiographs can provide useful diagnostic information in sick raptors that exhibit vomiting or changes in appetite or casting frequency, and may help guide treatment decisions of impacted birds. Careful consideration of substrate, enrichment items, and access to potential foreign material that could be ingested may be the best pre-emptive management strategy in captive raptors.

Following the explosion of the Deepwater Horizon MC252 oil rig in 2010, 319 live sea turtles exposed to crude oil and oil-dispersant (Corexit) combinations were admitted to rehabilitation centers for decontamination and treatment. Treatment of oiled sea turtles was guided by expected physiological and pathological effects of crude oil exposure extrapolated from studies in other species and from a single loggerhead sea turtle (Caretta caretta) study. While invaluable starting points, inherent limitations to extrapolation, and small sample size of the experimental exposure study, reduce their utility for clinical guidance and for assessing oil spill impacts. Effects of dispersants were not included in the previous experimental exposure study, and cannot be effectively isolated in the analysis of field data from actual spills. A terminal study of pivotal temperature of sex determination using eggs salvaged from doomed loggerhead nests provided an opportunity for an ancillary exposure study to investigate the acute effects of crude oil, dispersant, and a crude oil/dispersant combination in sea turtle hatchlings. Eggs were incubated at 27.2 – 30.8 °C, and hatchlings were randomly assigned to control, oil, dispersant, and combined oil/dispersant exposures for 1 day or 4 days. Contaminant exposures were started after a 3 day post-hatching period simulating nest emergence. Turtles were placed in individual glass bowls containing aged seawater and exposed to oil (Gulf Coast – Mixed Crude Oil Sweet, CAS #8002-05-9, 0.833 mL/L) and/or dispersant (Corexit 9500A, 0.083 mL/L), replicating concentrations encountered during oil spills and subsequent response. Statistically significant differences between treatments and non-exposed controls were detected for PCV, AST, uric acid, glucose, calcium, phosphorus, total protein, albumin, globulin, potassium and sodium. The principal dyscrasias reflected acute osmolar, electrolyte and hydration challenges that were more numerous and greater in combined oil/dispersant exposures at 4 days. Clinicopathological findings were supported by a failure to gain weight (associated with normal hatchling hydration in seawater) in dispersant and combination exposed hatchlings. These findings can help guide clinical response for sea turtles exposed to crude oil and crude oil/dispersant combinations, and indicate potential impacts on wildlife to consider when deploying dispersants in an oil spill response.

Enrofloxacin is a fluoroquinolone widely used in animals including fish. Intramuscular (IM) injection of enrofloxacin is a feasible and efficacious option for drug delivery. In many species IM injection has been associated with injection site reactions and increases in serum muscle enzymes. Injection site reactions have not been well characterized in fish. Three groups of striped bass (

OBJECTIVE: To determine the combined mydriatic effects of topical rocuronium bromide and phenylephrine in juvenile loggerhead turtles and identify any adverse effects associated with treatment. ANIMALS STUDIED: Eleven juvenile loggerhead turtles (Caretta caretta). PROCEDURES: Four 20 μL drops of rocuronium bromide and four 20 μL drops of 10% phenylephrine were placed into the right eye at 2‐minute intervals of 5 turtles, while the same volume of saline was administered to six control turtles. A pupilometer recorded pupil measurements at rest and following a light stimulus at 2, 15, 30, 60, 120, 150, 180, 210, 240, 300, and 360 minutes following delivery of the final drop to the ocular surface. Intraocular pressure (IOP) was also measured at similar time points. RESULTS: The nonilluminated and light‐stimulated pupillary diameter of the right eye of treated turtles was significantly greater than baseline starting at 120 and 15 minutes, respectively. Light‐stimulated pupillary diameter of treated eyes was greater than that of control eyes from time 15 minutes until the end of the treatment period. No systemic side effects were noted over a 24 hours period following treatment and all turtles showed normal behavior and appetite. No mydriasis was noted in either eye at 24 hours and the anterior segment was normal. CONCLUSIONS: A combination of topical ophthalmic rocuronium bromide and 10% phenylephrine is safe and effective for mydriasis in juvenile loggerhead turtles.

Background The Southern Stingray (Dasyatis americana) is a batoid elasmobranch frequently exhibited in zoological institutions. Blood is commonly collected from the caudal hemal arch at the tail base in stingrays for the purpose of health assessment and clinical pathology tests. An alternative site that allows a dorsal or ventral approach without necessitating puncture of a cartilaginous structure has been identified between the cartilaginous pectoral fin rays (ceratotrichia). Objectives The purpose of the study was to compare CBC, plasma biochemistry analytes, and blood gas variables between blood samples collected from the caudal and pectoral fin vasculature sites of the Southern Stingray. Methods Fifteen captive Southern Stingrays (10 females, 5 males) from 4 zoo and aquarium facilities were sampled. Lithium heparinized blood samples were collected from the caudal and pectoral venipuncture sites of each animal. Values from estimated total and differential leukocyte counts, plasma biochemistry analytes, and blood gas variables were compared. Results There were no statistically significant differences between venipuncture sites for the measured analytes except for CK activity, which was statistically significantly higher in the pectoral site samples. Levels of agreement between sites were good or moderate for 22 analytes and poor for ALT, AST, CK, pO2, lactate, monocytes, and eosinophils. Conclusions The good agreement between sampling sites for the majority of the measured analytes and the lack of differences that would alter clinical interpretation support the use of the pectoral site as an alternative to the traditional caudal fin venipuncture site in Southern Stingrays.

Intravascular access in batoid species is commonly achieved using the ventral coccygeal or radial wing vessels. However, these approaches can be difficult because of the presence of cartilage, lack of specific landmarks, species variation, and small vessel size in many species. This study used postmortem contrast radiography and gross dissection to develop landmarks for a new, dependable vascular access in three Myliobatiform species commonly maintained in captivity: Atlantic stingray (Hypanus sabinus), cownose ray (Rhinoptera bonasus), and smooth butterfly ray (Gymnura micrura). The mesopterygial vein provides quick vascular access and is suitable for administration of large fluid volumes and intravascular drugs. It is located immediately ventrolateral to the metapterygium cartilage, which sits adjacent to the coelomic cavity and supports the caudal half of the pectoral fin. Using the pectoral girdle and cranial third of the metapterygium cartilage as landmarks, vascular access can be achieved by directing a needle medially at approximately a 30° (adult cownose rays) or 45° angle (Atlantic stingrays, juvenile cownose rays, smooth butterfly rays) toward the metapterygium cartilage. Differences in the degree of needle direction are due to species and age-specific shapes of the metapterygium cartilage. The mesopterygial vein is an alternate site of quick and reliable venous access in batoid species.

Thromboelastography (TEG) provides a global evaluation of haemostasis. This diagnostic test is widely used in mammals but has not previously been performed in reptiles, mainly due to the limited availability of taxon-specific reagents. The objective of this pilot study was to establish a protocol to perform TEG in sea turtles. Pooled citrated plasma, stored at −80°C, from four green turtles (Chelonia mydas) was assayed on a TEG 5000. Several initiators were evaluated: kaolin (n=2), RapidTEG (n=2), fresh (n=2) and frozen (n=6) thromboplastin extracted from pooled brain tissue from several chelonian species, human recombinant tissue factor at 1:100 (n=1), Reptilase (n=2), and rabbit thromboplastin (n=1). Both fresh and frozen chelonian thromboplastin were superior in producing quantifiable TEG reaction time compared with all other reagents. These findings are consistent with the lack of an intrinsic pathway in turtles and confirmed a lack of coagulation in the turtle samples in response to mammalian thromboplastin. A TEG protocol was subsequently established for harvested species-specific frozen thromboplastin. The frozen thromboplastin reagent remained stable after one year of storage at −80°C. The developed protocol will be useful as a basis for future studies that aim to understand the pathophysiology of haemostatic disorders in various stranding conditions of sea turtles.

A 12 –yr-old female Goeldi's monkey (Callimico goeldii) in British Columbia, Canada was diagnosed with alveolar echinococcosis (AE) on postmortem examination. Echinococcus multilocularis has been identified in several species of nonhuman primates, most frequently Old World primates, in zoos and research facilities in Europe and Asia. The strain affecting this monkey was identified as a European haplotype, indistinguishable from E. multilocularis recently identified in several canids in British Columbia. The animal is suspected to have been exposed while living in a zoological institution in Alberta, where E. multilocularis has also been reported in urban coyotes. Alveolar echinococcosis is a zoonotic disease of increasing concern in the United States and Canada, and this disease should be considered on the differential list of any nonhuman primate exhibiting signs of abdominal pain or distension, along with diagnostic imaging consistent with cystic structures of the liver or other organs.

Measuring plasma protein electrophoresis fractions and plasma concentrations of acute phase proteins (APPs) is commonly used to monitor inflammation and illness in mammalian species, but its use is not widespread in fish species, despite the presence of similar proteins. This study aims to determine plasma protein electrophoresis values for koi carp ( Cyprinus carpio ) and to assess the use of plasma protein electrophoresis and commercially available assays for 3 primary APPs (C-reactive protein, serum amyloid A, and haptoglobin) as markers of inflammation in 13 koi following invasive surgery. Median packed cell volume decreased by nearly 50% but recovered to baseline by 14 days postsurgery. Plasma protein electrophoresis fractions were consistent across the experimental period and did not demonstrate an inflammatory pattern, but electrophoretograms exhibited variation between individual fish. No clinically meaningful changes were observed in the APPs of the test subjects 28 days following surgery. Despite grossly evident inflammation of the incision line, either this surgical procedure did not stimulate a marked systemic inflammatory response, or the available assays are not effective in detecting the acute phase inflammatory response in this fish species.

Leatherback sea turtles are globally distributed and endangered throughout their range. There are limited data available on disease in this species. Initial observations of solitary large intestinal diverticulitis in multiple leatherbacks led to a multi-institutional review of cases. Of 31 subadult and adult turtles for which complete records were available, all had a single exudate-filled diverticulum, as large as 9.0 cm in diameter, arising from the large intestine immediately distal to the ileocecal junction. All lesions were chronic and characterized by ongoing inflammation, numerous intralesional bacteria, marked attenuation of the muscularis, ulceration, and secondary mucosal changes. In three cases, Morganella morganii was isolated from lesions. Diverticulitis was unrelated to the cause of death in all cases, although risk of perforation and other complications are possible.

Cold-stunning in sea turtles is a frequent natural cause of mortality and is defined as a hypothermic state due to exposure to water temperatures

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.

Tricaine methanesulfonate (MS-222) solutions are used routinely for clinical and experimental anesthesia of a variety of aquatic organisms. It is commonly accepted that MS-222 alters pH when added to freshwater, and buffering freshwater anesthetic solutions is considered to be the standard of care. However, buffering saltwater solutions is considered by many to be unnecessary; some individuals question whether any water with high natural mineral hardness needs to be buffered when MS-222 is added. To assess the need for buffering, we studied the dynamics of pH change when adding clinically relevant amounts of MS-222 to artificial seawater and to freshwater of varying hardness. We found that artificial seawater does not possess adequate buffering capacity to prevent a physiologically significant drop in pH when MS-222 is added, and hardness does not accurately predict the reaction of water pH to the addition of MS-222 in clinically relevant concentrations. Our data for salt water suggest that MS-22...

Monogenean parasite infestations are common in captive marine teleosts, and are generally found on the skin and gills. This report describes an unusual pathological presentation of exuberant granulation tissue of the gills, suspected to be related to Neobenedenia infestation in two cobia housed together at a North Carolina aquarium.

Gonadectomy of pet lizards may be necessary to treat various medical conditions, and prophylactic surgery may prevent life-threatening reproductive-associated conditions. This study provides information relevant to the pre-surgical evaluation of bearded dragons (Pogona vitticeps). Blood samples collected from seven male and seven female inland bearded dragons prior to elective gonadectomy surgery, and again 12 days following surgery, were analyzed using a point-of-care blood gas analyzer (iStat) and a semiportable chemistry analyzer (Abaxis VetScan) to obtain plasma biochemistry and electrolyte concentrations. Statistically significant differences were seen for packed cell volume and protein, calcium, and potassium concentrations when pre-surgical values were compared with post-surgical values. Four female lizards in the cohort died in the immediate post-operative period, and three of these animals had large, well-developed ovarian follicles. The ovaries of all surviving female lizards were chara...