Effect of photobiomodulation therapy as a multimodal approach to treatment of dermal abrasions in cownose rays (Rhinoptera bonasus).

Photobiomodulation therapy (PBMT) has become increasingly utilized in the zoo and aquarium fields to treat ailments in various species; however, its use in teleosts and elasmobranchs is relatively underrepresented in the literature. This study examined the efficacy of PBMT as an adjunct treatment to reduce the healing time of dermal abrasions in cownose rays (Rhinoptera bonasus). Ten cownose rays were included in this study and separated into control (n = 5) and treatment groups (n = 5). Animals in both treatment and control groups received intramuscular injections of enrofloxacin every 72 h to provide broad-spectrum antibiotic coverage. Cownose rays in the treatment group additionally received PBMT using the Companion® CTC-12 unit using the large, noncontact head. Control group rays were subjected to identical handling without the use of PBMT. A total of 196 Joules were delivered at a power of 2 W based on a dose of 5 J/cm2 at each session. Observational analysis revealed a lack of appreciable difference in healing time between the treatment and control groups in the aforementioned settings; however, it is recommended that PBMT protocols should be re-evaluated if there is little to no response in healing after three to four sessions. PBMT use on acute superficial dermal abrasions in cownose rays was well-tolerated in this study and may have utility in a multimodal treatment approach to wound healing. This study also produced a wound scoring metric based on photographs captured at each treatment. Future studies should utilize increased fluence (J/cm2) and irradiance (W/cm2) and incorporate synchronous histological analysis and increased sample size.

RETROSPECTIVE REVIEW OF NEUROLOGIC DISEASE IN STRANDED ATLANTIC HARBOR SEALS (PHOCA VITULINA CONCOLOR) ALONG THE NEW ENGLAND COAST.

Harbor seals (Phoca vitulina) are a common species admitted to marine mammal rehabilitation facilities. As important indicators of marine ecosystem health, monitoring trends of disease in harbor seal populations is critical. However, few studies have evaluated neurologic disease in this species. The general objective of this study was to retrospectively review and delineate neurologic disease in free-ranging Atlantic harbor seals (P. vitulina concolor) that stranded along the New England (United States) coast and entered a rehabilitation facility between 2006 and 2019. Any Atlantic harbor seal that stranded live along the New England coast during the study period and was diagnosed with neurologic disease on either antemortem or postmortem evaluation was included; medical records and pathologic reports were reviewed. From 211 records, 24 animals met the inclusion criteria. Prevalence of neurologic disease was 11% in the study population and six major categories of neurologic disease were identified including: inflammatory (54%), idiopathic (33%), trauma (4%), congenital (4%), and degenerative (4%). Of the seals diagnosed with neurologic disease, 13 (54%) seals died during rehabilitation, 10 (42%) seals were euthanized, and 1 (4%) seal survived to release. Unique cases seen included a seal with Dandy-Walker-like malformation and another seal with histopathologic findings compatible with neuroaxonal dystrophy, a degenerative process that has not been previously reported in marine mammals. This study contributes to the overall knowledge of the health of free-ranging Atlantic harbor seals and may aid clinicians in characterizing neurologic conditions that may be present in seals undergoing rehabilitation.

INVESTIGATION OF THE USE OF SERUM BIOMARKERS FOR THE DETECTION OF CARDIAC DISEASE IN MARINE MAMMALS.

Cardiac disease has been extensively documented in marine mammals; however, it remains difficult to diagnose antemortem. Assays measuring cardiac troponin I (cTnI) and N-terminal pro-brain natriuretic peptide (NT-proBNP) are used as sensitive and specific biomarkers of cardiac disease in many species, but have not been widely investigated in marine mammals. This study aimed to provide a set of reference values for cTnI and NT-proBNP in belugas (BW) (Delphinapterus leucas), sea otters (SO) (Enhydra lutris), Steller sea lions (SSL) (Eumetopias jubatus), and California sea lions (CSL) (Zalophus californianus) with and without cardiac disease, and to determine if these biomarkers are useful indicators of cardiac disease in these species. First, existing immunoassays for cTnI and NT-proBNP were successfully validated utilizing species-specific heart lysate spiked serum. Cohorts were defined by histopathology as animals with no evidence of cardiac disease ("control"), with confirmed cardiac disease ("disease"), and with concurrent renal and cardiac disease ("renal") for which serum samples were then analyzed. Serum concentration ranges for cTnI (ng/ml) and NT-proBNP (pmol/L) were determined for control and disease cohorts. There was significantly higher cTnI (P= 0.003) and NT-proBNP (P= 0.004) concentrations in the CSL disease cohort, as well as positive trends in BW, SO, and SSL disease cohorts that did not reach statistical significance. NT-proBNP concentrations were significantly higher in the CSL renal cohort compared with the control (P < 0.001) and disease (P= 0.007) cohorts. These results suggest that cTnI and NT-proBNP may be clinically useful in the antemortem diagnosis of cardiac disease in CSL, and warrant further investigation in BW, SO, and SL.

BASELINE HEMATOLOGIC AND BIOCHEMICAL VALUES AND CORRELATIONS TO ENVIRONMENTAL PARAMETERS IN MANAGED JAPANESE SPIDER CRABS (MACROCHEIRA KAEMPFERI).

Although the evaluation of hematologic and biochemical parameters is a well-established diagnostic tool in vertebrate medicine, comprehensive understanding of these parameters in invertebrate species is lacking. This study provides baseline hemocyte concentrations and biochemistry values for a population of managed Japanese spider crabs (JSC; Macrocheira kaempferi) housed at six different public aquariums. The methodology for obtaining diagnostic hemolymph samples is described. Distinct hemocyte types were identified, including hyaline cells, semigranulocytes, and granulocytes, with hyaline cells as the predominant type. Correlates to exam findings and environmental parameters were evaluated and included higher absolute semigranulocyte counts (r = 0.65, P = 0.020) and triglyceride levels (r = 0.44, P = 0.014) in JSC with exoskeletal lesions; higher total protein (mean = 5.93 g/dl, P = 0.028), cholesterol (median = 18.5 mg/dl, P = 0.018), triglyceride (median = 15.5 mg/dl, P = 0.002), and amylase (median = 243 U/L, P = 0.013) in nonmolting JSC compared with JSC that have previously molted since acquisition (total protein mean = 4.83 g/dl, cholesterol median = 14 mg/dl, triglyceride median = 6.4 mg/dl, and amylase median = 131 U/L); and lower relative and absolute granulocyte counts (mean = 8.83% P = 0.030, median = 1,162 cells/µl P = 0.006, respectively) and higher albumin (median = 1.35 g/dl, P = 0.031) in JSC housed at facilities that used ozone sterilization. The data presented serve as a foundation for understanding basic clinical parameters in JSC hemolymph, as well as the potential influence of environmental stressors on those parameters.

Use of a canine melanoma vaccine in the management of malignant melanoma in an African penguin (Spheniscus demersus).

CASE DESCRIPTION: A 25-year-old 4.4-kg male aquarium-hatched African penguin (Spheniscus demersus) was evaluated because of a raised 1.5 × 0.5-cm pigmented mass extending from within the right naris noted 2 days earlier. CLINICAL FINDINGS: The penguin had a raised pigmented mass extending out from the right naris and onto the upper beak. Histologic examination of excisional biopsy specimens confirmed a diagnosis of malignant melanoma. A treatment plan including administration of meloxicam, radiation therapy, and immunotherapy was initiated. TREATMENT AND OUTCOME: Treatment with meloxicam (0.2 mg/kg, PO, q 24 h) was initiated and continued for a total of 45 weeks; however, the medication was discontinued for a period of 6 weeks because of the risk of toxic effects in the chick that the penguin was feeding at that time. The penguin underwent local hypofractionated radiation therapy and received 4 once weekly 8-Gy fractions of radiation (total radiation dose, 32 Gy). The penguin was administered a canine melanoma vaccine transdermally every other week for 4 doses, with a booster injection given 7 months after the first dose. Treatment with the vaccine appeared to have no adverse effects. The penguin's pre- and postvaccination tyrosinase-specific antibody titers were measured with an anti-human tyrosinase-specific ELISA, and a 3-fold titer increase indicated a positive humoral immune response to the canine melanoma vaccination. The penguin died of unrelated causes 54 weeks after initial diagnosis, and there was no evidence of metastasis on necropsy. CLINICAL RELEVANCE: These case findings suggested that vaccination with a canine melanoma vaccine may be a safe and useful adjunct treatment for management of malignant melanoma in penguins.

TOTAL EAR CANAL ABLATION AND LATERAL BULLA OSTEOTOMY (TECA-LBO) IN ATLANTIC HARBOR SEALS (PHOCA VITULINA CONCOLOR) FOR SUCCESSFUL SURGICAL MANAGEMENT OF OTITIS MEDIA.

Chronic, severe otitis media was diagnosed in four Atlantic harbor seals (Phoca vitulina concolor), three of which were stranded animals undergoing rehabilitation. All seals presented with unilateral purulent aural discharge that would intermittently recur despite prolonged topical and systemic antimicrobial therapy. Aerobic culture from aural discharge isolated multidrug-resistant organisms in all seals, including Pseudomonas aeruginosa, Staphylococcus pseudintermedius, Klebsiella pneumoniae, and/or Enterococcus faecalis. Computed tomography was used in three cases to confirm otitis media and positive contrast ear canalography was used in one case to confirm tympanic membrane rupture. Given the persistent nature of otitis, surgical intervention in the form of a total ear canal ablation and lateral bulla osteotomy (TECA-LBO) was indicated. Surgery was successful in achieving complete clinical resolution of otitis in all seals. Postoperative complications included temporary unilateral paralysis of the left nare (2/4) and a transient left ptosis (1/4). Partial to complete surgical site dehiscence occurred in all cases; however, complete healing was achieved by second intention in 60 d or less. One rehabilitated seal was fitted with a satellite tag that confirmed normal swimming and diving patterns post release. In harbor seals, TECA-LBO can be performed safely to treat persistent cases of otitis media and should be considered in cases of chronic otitis that are not responsive to medical management.

RADIOGRAPHIC DETERMINATION OF GASTRIC EMPTYING AND GASTROINTESTINAL TRANSIT TIME IN COWNOSE RAYS (RHINOPTERA BONASUS) AND WHITESPOTTED BAMBOO SHARKS (CHILOSCYLLIUM PLAGIOSUM) AND THE EFFECT OF METOCLOPRAMIDE ON GASTROINTESTINAL MOTILITY.

Gastrointestinal (GI) pathology is common in elasmobranchs; however, information regarding normal GI transit time and the effect of therapeutics on GI motility is lacking. The objective of this study was to determine baseline gastric emptying and GI transit times in cownose rays (Rhinoptera bonasus) and whitespotted bamboo sharks (Chiloscyllium plagiosum) via radiographic barium sulfate contrast studies. Additionally, a pilot study was undertaken to determine the effect of metoclopramide on GI transit time in whitespotted bamboo sharks. Eight cownose rays and eight whitespotted bamboo sharks were administered a 98% w/w barium sulfate suspension at 8 ml/kg via orogastric tube. Post-contrast radiographs were obtained at 2 min, 3, 6, 12, and 23 hr for rays; and 2 min, 3, 6, 9, 12, 16, 25, 30, 36, and every 12 hr until complete gastric emptying occurred for sharks. In cownose rays, the mean and standard error were established for time of initial spiral colon filling (3.4 ± 0.4 hr), complete spiral colon opacification (12 ± 0 hr), initial spiral colon emptying (21.6 ± 1.4 hr), and complete gastric emptying (23 ± 0 hr). In bamboo sharks, the mean and standard error were established for time of initial spiral colon filling (5.3 ± 0.5 hr), complete spiral colon opacification (12.4 ± 1.3 hr), initial spiral colon emptying (22.5 ± 2.7 hr), and complete gastric emptying (39.9 ± 3.3 hr). Cownose rays had a significantly shorter time to spiral colon filling and complete gastric emptying compared with bamboo sharks (P < 0.05). Whitespotted bamboo sharks (n = 8) were administered metoclopramide (0.4 mg/kg orally once daily for 10 days) and the barium series was repeated. Complete gastric emptying time was significantly shorter in treated sharks compared with control (P < 0.05), suggesting that metoclopramide may be a useful therapeutic for GI motility disorders in elasmobranchs.

SERUM ACUTE-PHASE PROTEINS IN BOTTLENOSE DOLPHINS (TURSIOPS TRUNCATUS) AND CORRELATION WITH COMMONLY UTILIZED INFLAMMATORY INDICES.

Acute-phase proteins (APP) are the foundation to the innate immune response and valuable biomarkers that increase with inflammation, infection, neoplasia, stress, and trauma.2,4,16 Little is known about the acute-phase response in cetaceans and if these proteins can be used for health monitoring in individuals and free-ranging populations. The purpose of this study was to characterize serum concentrations of haptoglobin (Hp) and serum amyloid A (SAA), as well as electrophoretic profiles of common bottlenose dolphins (Tursiops truncatus) in free-ranging (n = 33) and professional care (n = 27) settings. Results were correlated to commonly utilized inflammatory indices including erythrocyte sedimentation rate (ESR), fibrinogen, total white blood cell count (WBC), and absolute neutrophil count. SAA levels, measured with a dolphin-specific enzyme linked immunosorbent assay (ELISA), were significantly higher (P = 0.05) in free-ranging dolphins (mean = 4.26; SE = 1.12) when compared with those under professional care (mean = 1.82; SE = 0.45). For dolphins under professional care, a statistically significant correlation was identified between ESR and Hp (P < 0.001; r = 0.69), ESR and SAA (P < 0.001; r = 0.67), fibrinogen and Hp (P = 0.001; r = 0.58), and fibrinogen and SAA (P = 0.002; r = 0.56). In addition, there was a significant correlation between WBC and SAA (P = 0.01; r = 0.38) and absolute neutrophil count and SAA (P = 0.04; r = 0.32). There were no significant correlations between study variables observed in free-ranging dolphins. The variable correlation of APPs with commonly utilized inflammatory indices demonstrates that these proteins are independent measures of inflammation with unique sensitivity, specificity, and timeline of expression. The results of this study contribute to improved health monitoring of dolphins and have the potential to assist in identification of compromised health.

Labeled quantitative mass spectrometry to study the host response during aspergillosis in the common bottlenose dolphin (Tursiops truncatus).

Aspergillosis is a fungal infection caused by Aspergillus molds that can affect both humans and animals. Despite advances in diagnostics and therapy, medical management of this disease remains difficult. Expansion of the basic knowledge regarding its pathophysiology in animals is critical to aid in the identification of new biomarkers of infection for diagnosis and therapeutic targets. For such a purpose, proteomics can be used by addressing protein changes during various disease processes. In the present study, a mass spectrometry analysis based on isobaric tagging for relative and absolute quantitation (iTRAQ®) was applied for direct identification and relative quantitation of proteins in blood collected from 32 Aspergillus-diseased common bottlenose dolphins (Tursiops truncatus, 32 samples) in comparison with blood from 55 other dolphins (55 samples from 41 clinically-normal controls and from 14 cetaceans with miscellaneous non-Aspergillus inflammation diseases) and ten convalescent dolphins (28 samples). Sixty-six and 40 proteins were found to be ≥2.0-fold over- and underrepresented versus miscellaneous non-Aspergillus inflammatory dolphins, respectively, and most were confirmed vs. clinically-normal controls and convalescents. Many proteins which play a role in the adaptive immune response were identified, including MHC proteins and others involved in catalytic activity like the NADPH-ubiquinone oxido-reductases. Overall, iTRAQ® appears to be a convenient proteomic tool greatly suited for exploratory ex vivo studies focusing on pathophysiology. This technique should be considered as a preliminary step before validation of new diagnostic markers.

Prognostic indicators for survival in African penguin chicks (Spheniscus demersus): A retrospective analysis.

Mystic Aquarium has successfully maintained an active breeding population of African penguins since 1990. Between 1990 and 2015, 477 eggs were laid, 84 of which were recommended for rearing according to the SSP Breeding and Transfer Plan. Sixty-five percent hatched successfully, and total fertility ranged from 56% to 78%. Of the 55 hatchlings, 60% of chicks reached fledgling age (80 days). Of the 22 chicks that died before fledging, 59% died within 7 days of hatching, and 82% within 30 days of hatching. Categories of mortality were broken down into the following categories: failure to thrive (in the absence of a definitive cause of death) (41%), respiratory disease (32%), parent-associated trauma (13%), congenital defects (9%), and gastrointestinal disease (5%). Analysis of associated factors identified a significant correlation between the hatch weights of chicks that survived beyond day 7 (60.5-66.0 g), and those that did not (49.5-59.5 g). Further positive correlations were established between chick hatch weight and both dam body weight, and egg weight. While hand-reared chicks gained weight more slowly than their parent-reared counterparts, results showed that rearing method had no significant impact on survival. The findings of this study provide a baseline for assessing reproductive success and chick mortality, and supply prognostic indicators for evaluating chick success in managed African penguin populations.

Seroprevalence Survey for Microsporidia in Common Bottlenose Dolphin ( Tursiops truncatus): Example of a Quantitative Approach Based on Immunoblotting.

Little is known about microsporidiosis pathogenicity in cetaceans. Here we report seroprevalence of 76% for microsporidia in blood samples from common bottlenose dolphins ( Tursiops truncatus), from animals managed under human care ( n=108) or captured for health assessments ( n=13) and released.

BASELINE CORTICOSTERONE, HEMATOLOGY, AND BIOCHEMISTRY RESULTS AND CORRELATIONS TO REPRODUCTIVE SUCCESS IN NESTING LOGGERHEAD SEA TURTLES ( CARETTA CARETTA).

Characterizing the health status and reproductive success of wild populations of sea turtles can be difficult; however, obtaining data to do this can provide important insight into the stability and long-term success of a population. This study examined the use of baseline corticosterone to assess reproductive success of a population of nesting loggerhead sea turtles ( Caretta caretta) on Jekyll Island, Georgia and investigated hematological and biochemical trends in this population. A total of 37 nesting loggerhead sea turtles was sampled for this study. Eleven (29.7%) turtles were sampled in 2013 and 26 (70.3%) were sampled in 2014. A majority of the turtles sampled successfully nested (29/37, 78.4%; false crawls: 8/37, 21.6%). There was no significant correlation between baseline corticosterone concentrations and reproductive success (hatch success or emergence success) in this population. There was a significant positive correlation ( r = 0.461, P = 0.02) between corticosterone concentrations and absolute monocyte counts. There was a significant negative correlation between monocyte count and hatch success ( r = -0.464, P = 0.05) and a positive correlation between phosphorus and hatch success ( r = 0.405, P = 0.05). Calcium concentrations were significantly different ( P = 0.01) between animals that false crawled and those that nested, with nesting turtles having lower calcium concentrations than those that false crawled. Turtles that false crawled were significantly ( P = 0.008) more likely to have elevated potassium concentrations than turtles that nested. This study provides the first attempt at characterizing baseline corticosterone, hematology, and biochemistry data and correlations with reproductive success in nesting loggerhead sea turtles. Overall, loggerhead sea turtles capable of nesting were found to be in good health and have good reproductive success while maintaining low levels of corticosterone during reproductive activities.

Evaluation of a genus-specific ELISA and a commercial Aspergillus Western blot IgG® immunoblot kit for the diagnosis of aspergillosis in common bottlenose dolphins (Tursiops truncatus).

Aspergillosis is a fungal infection with high mortality and morbidity rates. As in humans, its definitive diagnosis is difficult in animals, and thus new laboratory tools are required to overcome the diagnostic limitations due to low specificity and lack of standardization. In this study of common bottlenose dolphins (Tursiops truncatus), we evaluated the diagnostic performance of a new commercial immunoblot kit that had been initially developed for the serologic diagnosis of chronic aspergillosis in humans. Using this in a quantitative approach, we first established its positive cutoff within an observation cohort of 32 serum samples from dolphins with "proven" or "probable" diagnosis of aspergillosis and 55 negative controls. A novel enzyme-linked immunosorbent assay (ELISA) test was also developed for detecting anti-Aspergillus antibodies, and results were compared between the two assays. Overall, the diagnostic performance of immunoblot and ELISA were strongly correlated (P < .0001). The former showed lower sensitivity (65.6% versus 90.6%), but higher specificity (92.7% vs. 69.1%), with no cross-reaction with other fungal infections caused by miscellaneous non-Aspergillus genera. When assessing their use in a validation cohort, the immunoblot kit and the ELISA enabled positive diagnosis before mycological cultures in 42.9% and 33.3% subjects addressed for suspicion of aspergillosis, respectively. There was also significant impact of antifungal treatment on the results of the two tests (P < .05). In all, these new serological methods show promise in aiding in the diagnosis of aspergillosis in dolphins, and illustrate the opportunity to adapt commercial reagents directed for human diagnostics to detect similar changes in other animals.

CUTANEOUS SQUAMOUS CELL CARCINOMA IN A PANTHER CHAMELEON (FURCIFER PARDALIS) AND TREATMENT WITH CARBOPLATIN IMPLANTABLE BEADS.

A 3-yr-old male panther chameleon (Furcifer pardalis) presented with bilateral raised crusted skin lesions along the lateral body wall that were found to be carcinoma in situ and squamous cell carcinoma. Similar lesions later developed on the caudal body wall and tail. A subcutaneous implantable carboplatin bead was placed in the first squamous cell carcinoma lesion identified. Additional new lesions sampled were also found to be squamous cell carcinomas, and viral polymerase chain reaction was negative for papillomaviruses and herpesviruses. Significant skin loss would have resulted from excision of all the lesions, so treatment with only carboplatin beads was used. No adverse effects were observed. Lesions not excised that were treated with beads decreased in size. This is the first description of cutaneous squamous cell carcinoma and treatment with carboplatin implantable beads in a panther chameleon.

CIRCULATING CONCENTRATIONS OF THYROID HORMONE IN BELUGA WHALES (DELPHINAPTERUS LEUCAS): INFLUENCE OF AGE, SEX, AND SEASON.

Thyroid hormones play a critical physiologic role in regulating protein synthesis, growth, and metabolism. To date, because no published compilation of baseline values for thyroid hormones in beluga whales (Delphinapterus leucas) exists, assessment of thyroid hormone concentrations in this species has been underused in clinical settings. The purpose of this study was to document the concentrations of total thyroxine (tT4) and total triiodothyronine (tT3) in healthy aquarium-maintained and free-ranging beluga whales and to determine the influence of age, sex, and season on the thyroid hormone concentrations. Archived serum samples were collected from healthy aquarium-maintained (n=43) and free-ranging (n=39) belugas, and serum tT4 and tT3 were measured using chemiluminescence immunoassay. The mean tT4 concentration in aquarium-maintained belugas was 5.67±1.43 µg/dl and the mean tT3 concentration was 70.72±2.37 ng/dl. Sex comparisons showed that aquarium-maintained males had significantly greater tT4 and tT3 (9.70±4.48 µg/dl and 92.65±30.55 ng/dl, respectively) than females (7.18±2.82 µg/dl and 77.95±20.37 ng/dl) (P=0.004 and P=0.013). Age comparisons showed that aquarium-maintained whales aged 1-5 yr had the highest concentrations of tT4 and tT3 (8.17±0.17 µg/dl and 105.46±1.98 ng/dl, respectively) (P=0.002 and P<0.001). tT4 concentrations differed significantly between seasons, with concentrations in winter (4.59±1.09 µg/dl) being significantly decreased compared with spring (P=0.009), summer (P<0.0001), and fall (P<0.0001) concentrations. There was a significant difference in tT4 and tT3 concentrations between aquarium-maintained whales (5.67±1.43 µg/dl and 70.72±15.57 ng/dl, respectively) and free-ranging whales (11.71±3.36 µg/dl and 103.38±26.45 ng/dl) (P<0.0001 and P<0.001). Clinicians should consider biologic and environmental influences (age, sex, and season) for a more accurate interpretation of thyroid hormone concentrations in belugas. The findings of this study provide a baseline for thyroid health monitoring and comprehensive health assessments in both aquarium-maintained and free-ranging beluga whales.

Baseline plasma corticosterone, haematological and biochemical results in nesting and rehabilitating loggerhead sea turtles (Caretta caretta).

The evaluation of hormonal responses to stress in reptiles relies on acquisition of baseline corticosterone concentrations; however, the stress associated with the restraint needed to collect the blood samples can affect the results. The purpose of this study was to determine a time limit for the collection of blood samples to evaluate baseline corticosterone, haematological and biochemical results in nesting (n = 11) and rehabilitating (n = 16) loggerhead sea turtles (Caretta caretta). Blood samples were collected from the dorsal cervical sinus of each turtle immediately after touching the animal (t 0; 0-3 min) and 3 (t 3; 3-6 min), 6 (t 6; 6-9 min; nesting turtles only), 10 (t 10; 10-13 min) and 30 min (t 30; rehabilitating turtles only) after the initial hands-on time. Consistent between the rehabilitating and nesting turtles, there was a subtle yet significant increase in white blood cell counts over time. Despite the fact that white blood cell counts increased during the sampling period, there was no direct correlation between white blood cell count and corticosterone in the sampled turtles. In the nesting turtles, significant elevations in corticosterone were noted between t 0 and t 3 (P = 0.014) and between t 0 and t 6 (P = 0.022). Values at t 10 were not significantly different from those at t 0 (P = 0.102); however, there was a trend for the corticosterone values to continue to increase. These results suggest that sampling of nesting loggerhead sea turtles within 3 min of handling will provide baseline corticosterone concentrations in their natural environment. Significant elevations in corticosterone were also noted in the rehabilitating loggerhead sea turtles between t 0 and t 10 (P = 0.02) and between t 0 and t 30 of sampling (P = 0.0001). These results suggest that sampling of loggerhead sea turtles within 6 min of handling should provide baseline corticosterone concentrations in a rehabilitation setting. The delay in the corticosterone response noted in the rehabilitating turtles may be associated with the daily contact (visual or direct) they have with their human caretakers.

Esophageal squamous cell carcinoma in six harbor seals (Phoca vitulina spp.).

Six cases of esophageal squamous cell carcinoma were identified in six captive adult Pacific (Phoca vitulina richardsii; n = 2) and Atlantic (Phoca vitulina concolor; n = 4) harbor seals. These seals presented with intermittent dysphagia, regurgitation, inappetence, and abnormal posturing. Common clinical pathology findings in these seals included azotemia, hyperproteinemia, hyperglobulinemia, and leukocytosis. Gastrointestinal endoscopy commonly revealed an ulcerated mass near the gastroesophageal junction. Each seal was euthanized (n = 3) due to poor prognosis, subsequently died while undergoing an anesthetic procedure (n = 2), or found dead (n = 1). The diagnosis of squamous cell carcinoma was confirmed via biopsy of esophageal mucosa during endoscopy or histopathologic examination of affected tissues after necropsy. On the basis of clinical and postmortem findings, esophageal squamous cell carcinoma should be considered as a differential diagnosis in aged harbor seals exhibiting clinical signs of regurgitation, decreased appetite or anorexia, vomiting, and/or abnormal posturing.

Plasma electrophoretic profiles and hemoglobin binding protein reference intervals in the eastern box turtle (Terrapene carolina carolina) and influences of age, sex, season, and location.

Evaluation of plasma electrophoretic profiles and acute phase protein concentrations may play a valuable role in health assessment of reptiles; however, little is known about reference intervals in free-ranging eastern box turtles (Terrapene carolina carolina). The purpose of this study was to establish reference intervals of protein electrophoretic profiles and hemoglobin binding protein ([HBP] as determined by a haptoglobin assay) in free-ranging eastern box turtles and to assess any possible correlations between varying age class (adults vs. juvenile), sex (male, female, or unknown), season (spring, summer, or fall), or location (Tennessee vs. Illinois). Blood samples were obtained from 324 eastern box turtles from 2010 to 2012 at three sites in Illinois and one site in Tennessee, USA. Significant differences were observed with total protein (sex, season, state, Illinois location), albumin (age class, season, state, Illinois location), α-1 globulins (sex, season, Illinois location), α-2 globulins (sex, season, state, Illinois location), ß globulins (age class, sex, season, state, Illinois location), γ globulins (sex, season state, Illinois location), and hemoglobin binding protein (age class, sex, state, Illinois location). The use of electrophoretic profiles and acute phase proteins is a relatively new concept in reptilian medicine, and this study allowed for establishment of references intervals in the eastern box turtle and emphasized differences that occured based on age, sex, season, and location. Future research in this area can now build on these data to determine changes in population health over time or alterations due to specific environmental or disease threats.

Antemortem diagnosis and successful management of noncompressive segmental myelopathy in a Siberian-Bengal mixed breed tiger.

A 10-yr-old female spayed mixed breed tiger presented for a 9-day history of acute and nonprogressive paralysis of the pelvic limbs. Magnetic resonance imaging revealed a lesion suggestive of fibrocartilaginous embolic myelopathy with regional spinal cord edema, decreased disk signal intensity at L2-L3, and mild intervertebral disk protrusion at L1-L2 and L2-L3. Cerebral spinal fluid analysis showed no overt evidence of infection or neoplasia. Medical therapy was instituted, including corticosteroids and gastroprotectants as well as nursing care and physical therapy. The tiger began showing clinical improvement 2 wk after initiating treatment, progressing to the point where the animal was standing and intermittently walking. Three months after diagnosis, the tiger had regained muscle strength of its hind limbs and walked regularly with improving coordination. This case is the first report of antemortem diagnosis and successful medical management of suspected fibrocartilaginous embolic myelopathy in a large exotic felid.