Physiologic responses of Cownose Rays following freshwater dips for treatment of capsalid monogeneans.

OBJECTIVE: Two Cownose Rays Rhinoptera bonasus were presented for management of a severe capsalid monogenean infection Benedeniella posterocolpa in a mixed species habitat at an aquarium. METHODS: A series of freshwater (FW) dips were elected to mitigate parasite resistance to praziquantel due to the endemic monogenean population in the system. A pretreatment blood sample was opportunistically obtained, and subsequent samples were processed due to development of clinical signs. RESULT: While the first FW dip was tolerated well and did not induce abnormal behavior, the second led to one individual presenting with tachypnea, pallor, and lethargy. Marked hematological abnormalities requiring medical intervention occurred in both individuals after the second dip. After treatment with elasmobranch Ringer's fluid therapy, sodium bicarbonate, prednisolone acetate, and vitamin E/selenium in the more severely affected ray, and elasmobranch Ringer's alone in the second ray, hematologic derangements and symptoms resolved. Hemoconcentration, hyperproteinemia, hyperlactatemia, and hyperglycemia were attributed to a stress response. A selective loss of urea after exposure to FW resulted in decreased plasma osmolality. CONCLUSION: Given the severity of the stress response and associated complications observed, hyposalinity treatments should be utilized with caution in this species. If this therapy is employed, the clinician is advised to be prepared to administer supportive care coinciding with the FW dip if necessary.

The microbiome of African penguins (Spheniscus demersus) under managed care resembles that of wild marine mammals and birds.

Animals under managed care in zoos and aquariums are ideal surrogate study subjects for endangered species that are difficult to obtain in the wild. We compared the fecal and oral microbiomes of healthy, managed African penguins (Spheniscus demersus) to those of other domestic and wild vertebrate hosts to determine how host identity, diet, and environment shape the penguin microbiome. The African penguin oral microbiome was more similar to that of piscivorous marine mammals, suggesting that diet and a marine environment together play a strong role in shaping the oral microbiome. Conversely, the penguin cloaca/fecal microbiome was more similar to that of other birds, suggesting that host phylogeny plays a significant role in shaping the gut microbiome. Although the penguins were born under managed care, they had a gut microbiome more similar to that of wild bird species compared to domesticated (factory-farmed) birds, suggesting that the managed care environment and diet resemble those experienced by wild birds. Finally, the microbiome composition at external body sites was broadly similar to that of the habitat, suggesting sharing of microbes between animals and their environment. Future studies should link these results to microbial functional capacity and host health, which will help inform conservation efforts.

RETROSPECTIVE REVIEW OF PROPOFOL ANESTHESIA IN MULTIPLE ELASMOBRANCH SPECIES AT GEORGIA AQUARIUM, 2010-2022.

General anesthesia is often required in elasmobranch species for medical procedures. A variety of anesthetic drugs have been administered to elasmobranchs with wide variability in efficacy and safety. A retrospective review of 47 anesthetic procedures using IV propofol in eight different elasmobranch species at the Georgia Aquarium from 2010 to 2022 was performed. Cases involving seven sand tiger sharks (Carcharias taurus), four largetooth sawfish (Pristis perotteti), one longcomb sawfish (Pristis zijsron), four blacktip reef sharks (Carcharhinus melanopterus), three silvertip sharks (Carcharhinus albimarginatus), one sandbar shark (Carcharhinus plumbeus), five cownose rays (Rhinoptera bonasus), and one blotched fantail stingray (Taeniura meyeni) were evaluated. Induction dose of IV propofol (median: 2.5; 25-75%: 2.3-3.0; range: 1.7-4.0 mg/kg), time to desired effect (median: 4.0; 25-75%: 2.0-5.0; range: 0.5-15.0 min) and anesthetic duration (median: 76.0; 25-75%: 61.5-119.0; range: 27-216 min) were reported in all species. In six procedures (12.7%), maintenance of desired anesthetic plane required a supplemental dose of propofol IV (1 mg/kg) or addition of tricaine methanesulfonate (70 mg/L) as an immersion bath. The most common side effects were apnea and prolonged recovery. The IV propofol was efficacious and provided a procedural plane of anesthesia for a clinically relevant period of time in the majority of elasmobranch species, but observation for and management of complications is warranted.

BARIUM SULFATE GELATIN AS A NOVEL APPROACH TO FACILITATE GASTROINTESTINAL TRACT POSITIVE CONTRAST STUDIES IN A CAPTIVE HARBOR SEAL (PHOCA VITULINA) AND CALIFORNIA SEA LIONS (ZALOPHUS CALIFORNIANUS) WITHOUT RESTRAINT.

Gastrointestinal tract contrast medium studies are a valuable diagnostic modality to evaluate gastrointestinal anatomy, motility, and pathology. Four positive contrast medium studies were performed twice on a harbor seal (Phoca vitulina) and once each on two California sea lions (Zalophus californianus) to evaluate for gastrointestinal abnormalities by using barium-impregnated gelatin. Because marine mammals in human care routinely receive plain gelatin as a component of their diet and as secondary reinforcement, feeding a barium gelatin mitigates the need for tube feeding or restraint. Operant conditioning was used to position the pinnipeds for voluntary radiographs. The barium gelatin permitted adequate evaluation of gastric emptying and intestinal transit times and good evaluation of the structure of the intestinal tract. Full evaluation of gastric anatomy was limited due to the gelatin blocks not conforming to the stomach in their solid form and the barium quickly exiting the stomach as barium gelatin blocks disintegrated and because orthogonal views could not always be acquired. Even with these limitations, barium gelatin resulted in diagnostically valuable contrast imaging in a stress-free patient setting with reduced risk of aspiration and eliminated the effects that anesthesia can have on gastric motility as well as other anesthesia-associated risks.

Baseline Health and Nutritional Parameters of Wild Sand Tigers Sampled in Delaware Bay.

Species-specific hematological reference values are essential for diagnosis and treatment of disease and maintaining overall health of animals. This information is lacking for many species of elasmobranchs maintained in zoos and aquaria, thus reducing the effectiveness of care for these animals. Descriptive statistics and reference intervals were calculated for hematocrit and complete blood cell counts, biochemistry and protein electrophoresis parameters, trace minerals, vitamins, heavy metals, reproductive hormones, and fatty acids in the blood of 153 wild Sand Tigers Carcharias taurus of both sexes and a range of sizes caught in Delaware Bay (Delaware, USA). Mean hematocrit, total white blood cell counts, lymphocyte differentials, glucose, phosphorus, amylase, and aspartate aminotransferase levels were significantly higher in juveniles than in adults. Levels of estradiol, progesterone, testosterone, and differences in selenium and eicosapentaenoic acid (a polyunsaturated fatty acid) between males and females suggest that they are important parameters for improving Sand Tiger breeding success in managed care. Finally, blood metal levels for arsenic, cadmium, lead, and mercury suggest low levels of contaminant exposure for Sand Tigers during their summer residence in Delaware Bay. The results of this study provide baseline health parameters for wild Sand Tigers that will aid in effective maintenance of aquarium animals and contribute to a greater understanding of the biology of these sharks and efforts to accomplish sustainable management of their populations.

A novel herpes-like virus inducing branchial lesions in a tiger shark (Galeocerdo cuvier).

A juvenile, male tiger shark (Galeocerdo cuvier) developed illness after capture in Florida waters and was euthanized. Gross lesions included mild skin abrasions, hepatic atrophy, and coelomic fluid. Histologically, gills contained multifocal lamellar epithelial cell necrosis and thromboses. Scattered gill and esophageal epithelial cells had large, basophilic, intracytoplasmic, and intranuclear inclusions. Ultrastructurally, lamellar epithelial cells contained arrays of intracytoplasmic viral particles and scattered intranuclear nucleocapsids. Capsulated virions were 148 ± 11 nm with an 84 ± 8 nm icosahedral nucleocapsid and an electron-dense core. Next-generation sequencing, quantitative polymerase chain reaction, and in situ hybridization performed on formalin-fixed tissue confirmed a herpes-like viral infection. The viral polymerase shared 24% to 31% protein homology with other alloherpesviruses of fish, indicating a divergent virus. This report documents the pathologic findings associated with a molecularly confirmed novel herpes-like virus in an elasmobranch.

Multi-Injection Pharmacokinetics of Meloxicam in Kemp's Ridley (Lepidochelys kempii) and Green (Chelonia mydas) Sea Turtles after Subcutaneous Administration.

The objective of this study was to determine the pharmacokinetics and safety of multiple injections of meloxicam (MLX) administered subcutaneously (SQ) in Kemp's ridley (Lepidochelys kempii) and green (Chelonia mydas) sea turtles. Based on results from a previously published single-injection study, a multiple-injection regimen was derived for the Kemp's ridleys, which consisted of administering MLX at a dose of 1 mg/kg SQ every 12 h for 5 days, and for green turtles at a dose of 1 mg/kg SQ every 48 h for three treatments. Six turtles of each species were used for the study, and blood samples were taken at multiple time intervals. The terminal half-life after the last dose for the Kemp's ridley sea turtles was calculated at 7.18 h, and for the green sea turtles at 23.71 h. Throughout the multiple injections, MLX concentrations remained above 0.57 µg/mL, a concentration targeted in humans for the analgesic and anti-inflammatory effects. No negative side effects or changes to blood parameters evaluated were observed during the study in either species. The results of this study suggest MLX should be administered SQ to Kemp's ridley sea turtles at a dosage of 1 mg/kg every 12 h and in green sea turtles at a dose of 1 mg/kg every 48 h. The novelty of this work is that it is a multiple-injection study. Multiple injections were administered and produced concentrations that were considered therapeutic in humans, and the turtles did not have any adverse side effects. Furthermore, there were large differences in the pharmacokinetic values between green and Kemp's ridley sea turtles.

PHARMACOKINETIC BEHAVIOR OF MELOXICAM IN LOGGERHEAD (CARETTA CARETTA), KEMP'S RIDLEY (LEPIDOCHELYS KEMPII), AND GREEN (CHELONIA MYDAS) SEA TURTLES AFTER SUBCUTANEOUS ADMINISTRATION.

The objective of this study was to determine the pharmacokinetics of a single dose of meloxicam administered subcutaneously (SQ) to three species of sea turtles: loggerheads (Caretta caretta), Kemp's ridley (Lepidochelys kempii), and greens (Chelonia mydas). A dose of 1 mg/kg was given to the Kemp's ridleys and greens, whereas the loggerheads received 2 mg/kg. After SQ administration, the half-life (t1/2) of meloxicam administered at 1 mg/kg in the Kemp's ridleys was 5.51 hr but could not be determined in the greens. The half-life of meloxicam administered at 2 mg/kg in the loggerheads was 2.99 hr. The maximum concentration (Cmax) for meloxicam after SQ administration at 1 mg/kg in the Kemp's ridleys was 6.76 µg/ml and in the greens was 9.35 µg/ml. The Cmax in loggerheads for meloxicam after SQ administration at 2 mg/kg was 3.63 µg/mL. Meloxicam administered SQ at a dose of 1 mg/kg to the Kemp's ridley and greens provided measurable plasma concentrations of meloxicam for 48 and 120 hr, respectively, with no adverse side effects. In loggerheads, meloxicam administered SQ at a dose of 2 mg/kg provided measurable plasma levels of meloxicam for only 24 hr. Plasma levels of meloxicam of greater than 0.5 µg/ml are considered to be therapeutic in humans. Results suggested that administration of meloxicam SQ at 1 mg/kg in Kemp's ridleys and greens would result in plasma concentrations greater than 0.5 µg/ml for 12 and 120 hr, respectively. The administration of 2 mg/kg meloxicam to loggerhead turtles resulted in plasma concentrations greater than 0.5 µg/ml for only 4 hr.

Chronic exposure to glyphosate in Florida manatee.

Florida manatees depend on freshwater environments as a source of drinking water and as warm-water refuges. These freshwater environments are in direct contact with human activities where glyphosate-based herbicides are being used. Glyphosate is the most used herbicide worldwide and it is intensively used in Florida as a sugarcane ripener and to control invasive aquatic plants. The objective of the present study was to determine the concentration of glyphosate and its breakdown product, aminomethylphosphonic acid (AMPA), in Florida manatee plasma and assess their exposure to manatees seeking a warm-water refuge in Crystal River (west central Florida), and in South Florida. We analyzed glyphosate's and AMPA's concentrations in Florida manatee plasma (n = 105) collected during 2009-2019 using HPLC-MS/MS. We sampled eight Florida water bodies between 2019 and 2020, three times a year: before, during and after the sugarcane harvest using grab samples and molecular imprinted passive Polar Organic Chemical Integrative Samplers (MIP-POCIS). Glyphosate was present in 55.8% of the sampled Florida manatees' plasma. The concentration of glyphosate has significantly increased in Florida manatee samples from 2009 until 2019. Glyphosate and AMPA were ubiquitous in water bodies. The concentration of glyphosate and AMPA was higher in South Florida than in Crystal River, particularly before and during the sugarcane harvest when Florida manatees depend on warm water refuges. Based on our results, Florida manatees were chronically exposed to glyphosate and AMPA, during and beyond the glyphosate applications to sugarcane, possibly associated with multiple uses of glyphosate-based herbicides for other crops or to control aquatic weeds. This chronic exposure in Florida water bodies may have consequences for Florida manatees' immune and renal systems which may further be compounded by other environmental exposures such as red tide or cold stress.

Seasonal patterns in stable isotope and fatty acid profiles of southern stingrays (Hypanus americana) at Stingray City Sandbar, Grand Cayman.

Ecotourism opportunities in the marine environment often rely heavily on provisioning to ensure the viewing of cryptic species by the public. However, intentional feeding of wildlife can impact numerous aspects of an animals' behavior and ecology. Southern stingrays (Hypanus americana) provisioned at Stingray City Sandbar (SCS) in Grand Cayman have altered diel activity patterns and decreased measures of health. This study looked at seasonal changes in stable isotope (SI) and fatty acid (FA) profiles of provisioned stingrays at SCS. Plasma δ15N was higher in male stingrays (11.86 ± 1.71‰) compared to females (10.70 ± 1.71‰). Lower values for δ15N in males and females were measured in October during low tourist season, suggesting stingrays may be forced to rely on native prey items to supplement the decreased amount of provisioned squid available during this time. Plasma FA profiles were significantly different between sexes and across sampling time points, with FAs 22:6n3, 16:0, 20:5n3, 18:1n3C, 18:0 and 18:1n9T contributing to dissimilarity scores between groups. Dietary FAs primarily contributed to differences between males and females lending further evidence to differences in foraging patterns at SCS, likely due to intraspecific competition. Further, canonical analysis of principal coordinates (CAP) analysis of FA profiles suggest similar diets during peak tourist season and differences in diet between males and females during the low season. This study demonstrates alterations in feeding ecology in stingrays at SCS which is of critical importance for effective management of the SCS aggregation.

Natural environmental conditions and collaborative efforts provide the secret to success for sand tiger shark Carcharias taurus reproduction in aquaria.

Sand tiger sharks are an iconic large shark species held in aquaria worldwide. They rarely reproduce under managed care, with only seven aquaria reporting limited and sporadic success. For the first time in the Americas, a full-term young was born in an aquarium. The young was the result of breeding among a group of sharks purposefully brought together in 2016 for reproduction. Sharks were maintained in natural seawater and exposed to natural light and seasonal temperature fluctuations similar to their in situ counterparts. Decreased food consumption associated with breeding season and gestation was observed. Gestation time estimated from breeding observations and parturition was 321 days. Although the neonate was stillborn, this was a significant achievement. The husbandry details described within will be useful for other aquaria striving to support the reproduction of sand tiger sharks.

Primary intracranial fibrosarcoma in a southern sea otter Enhydra lutris nereis.

Southern sea otters Enhydra lutris nereis, a threatened marine mammal species, face numerous environmental and infectious disease challenges in their native habitat of coastal California, USA. However, there are few published cases describing neoplasia in sea otters despite their relatively long life span when cared for in aquarium settings. An 18 yr old neutered male southern sea otter, born and raised in human care, presented with an acute onset of seizures and dull mentation. Magnetic resonance imaging of the head revealed a large, central brain lesion. After no improvement with treatment, euthanasia was elected due to a poor prognosis. Grossly, a poorly demarcated, granular, tan mass expanded the cranial meninges in the longitudinal fissure at the level of the cruciate sulcus and extended into the underlying gray matter and superficial white matter. Histologically, the mass was composed of spindle cells, forming haphazardly arranged interlacing bundles and herringbone patterns, with a high mitotic count, moderate cellular pleomorphism, and prominent vascularization. Neoplastic cells demonstrated positive immunoreactivity for vimentin and negative immunoreactivity for smooth muscle actin, factor VIII-related antigen, S100, melan-A, E-cadherin, desmin, glial fibrillary acidic protein, and cytokeratin AE1/AE3. Based on gross, histologic, and immunohistochemical findings, the mass was most consistent with a primary intracranial fibrosarcoma (PIF). PIFs are a rare neoplasm in both humans and other animals with few reports in the veterinary literature. This is the first recorded case of a PIF in a sea otter.

Skin microbiome of beluga whales: spatial, temporal, and health-related dynamics.

BACKGROUND: Host-specific microbiomes play an important role in individual health and ecology; in marine mammals, epidermal microbiomes may be a protective barrier between the host and its aqueous environment. Understanding these epidermal-associated microbial communities, and their ecological- or health-driven variability, is the first step toward developing health indices for rapid assessment of individual or population health. In Cook Inlet, Alaska, an endangered population of beluga whales (Delphinapterus leucas) numbers fewer than 300 animals and continues to decline, despite more than a decade of conservation effort. Characterizing the epidermal microbiome of this species could provide insight into the ecology and health of this endangered population and allow the development of minimally invasive health indicators based on tissue samples. RESULTS: We sequenced the hypervariable IV region of bacterial and archaeal SSU rRNA genes from epidermal tissue samples collected from endangered Cook Inlet beluga whales (n = 33) and the nearest neighboring population in Bristol Bay (n = 39) between 2012 and 2018. We examined the sequences using amplicon sequence variant (ASV)-based analyses, and no ASVs were associated with all individuals, indicating a greater degree of epidermal microbiome variability among beluga whales than in previously studied cetacean species and suggesting the absence of a species-specific core microbiome. Epidermal microbiome composition differed significantly between populations and across sampling years. Comparing the microbiomes of Bristol Bay individuals of known health status revealed 11 ASVs associated with potential pathogens that differed in abundance between healthy individuals and those with skin lesions or dermatitis. Molting and non-molting individuals also differed significantly in microbial diversity and the abundance of potential pathogen-associated ASVs, indicating the importance of molting in maintaining skin health. CONCLUSIONS: We provide novel insights into the dynamics of Alaskan beluga whale epidermal microbial communities. A core epidermal microbiome was not identified across all animals. We characterize microbial dynamics related to population, sampling year and health state including level of skin molting. The results of this study provide a basis for future work to understand the role of the skin microbiome in beluga whale health and to develop health indices for management of the endangered Cook Inlet beluga whales, and cetaceans more broadly.

Testosterone and semen seasonality for the sand tiger shark Carcharias taurus†.

Understanding the fundamental reproductive biology of a species is the first step toward identifying parameters that are critical for reproduction and for the development of assisted reproductive techniques. Ejaculates were collected from aquarium (n = 24) and in situ (n = 34) sand tiger sharks Carcharias taurus. Volume, pH, osmolarity, sperm concentration, motility, status, morphology, and plasma membrane integrity were assessed for each ejaculate. Semen with the highest proportion of motile sperm was collected between April and June for both in situ and aquarium sand tiger sharks indicating a seasonal reproductive cycle. Overall, 17 of 30 semen samples collected from aquarium sharks from April through June contained motile sperm compared to 29 of 29 of in situ sharks, demonstrating semen quality differences between aquarium and in situ sharks. Sperm motility, status, morphology, and plasma membrane integrity were significantly higher (P < 0.05) for in situ compared to aquarium sand tiger sharks. Testosterone was measured by an enzyme immunoassay validated for the species. Testosterone concentration was seasonal for both aquarium and in situ sharks with highest concentrations measured in spring and lowest in summer. In situ sharks had higher (P < 0.05) testosterone concentration in spring than aquarium sharks. This study demonstrated annual reproduction with spring seasonality for male sand tiger sharks through marked seasonal differences in testosterone and semen production. Lower testosterone and poorer semen quality was observed in aquarium sharks likely contributing to the species' limited reproductive success in aquariums.

Comparing progesterone in blubber and serum to assess pregnancy in wild beluga whales (Delphinapterus leucas).

The Cook Inlet population of beluga whales (Delphinapterus leucas) is listed as endangered and continues to decline for largely unknown reasons; however, there is some evidence that poor reproductive success is a contributing factor. Pregnancy is difficult to detect through observation, and, there is reluctance to capture endangered beluga whales for reproductive tract imaging via ultrasound or to obtain suitable samples for pregnancy assessments. An endocrine analysis of blubber biopsies collected by remote darting could represent a minimally invasive way to identify pregnant females and compare pregnancy rates among years or populations. Studies have validated the use of blubber biopsies to identify pregnant females in other cetacean species, but not beluga whales; therefore, validation of blubber progesterone levels to proven tests that reliably detect pregnancy was needed for this species. As part of a larger study, we sampled blood and blubber from live-captured beluga whales (21 females, 26 males) in Bristol Bay, Alaska. Progesterone levels were determined in serum samples obtained from all animals and in blubber samples from a subset (14 females, 13 males) to determine pregnancy status, estimate the stage of pregnancy, and evaluate the suitability of using blubber alone for these assessments. In general, there was distinct separation of high levels of progesterone in serum and blubber for presumed pregnant females and low levels for males and presumed non-pregnant females. Blubber progesterone levels in two females (14% of females tested) were intermediate (i.e. ambiguous); their corresponding serum levels were consistent with being pregnant in one case and not being pregnant in the other. Except for these two intermediate values, pregnancy status of beluga whales could be determined from blubber alone, thereby providing a valuable tool to better understand reproduction dynamics from populations that cannot be captured for examination.

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.

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.

De novo transcriptome assembly and RNA-Seq expression analysis in blood from beluga whales of Bristol Bay, AK.

Assessing the health of marine mammal sentinel species is crucial to understanding the impacts of environmental perturbations on marine ecosystems and human health. In Arctic regions, beluga whales, Delphinapterus leucas, are upper level predators that may serve as a sentinel species, potentially forecasting impacts on human health. While gene expression profiling from blood transcriptomes has widely been used to assess health status and environmental exposures in human and veterinary medicine, its use in wildlife has been limited due to the lack of available genomes and baseline data. To this end we constructed the first beluga whale blood transcriptome de novo from samples collected during annual health assessments of the healthy Bristol Bay, AK stock during 2012-2014 to establish baseline information on the content and variation of the beluga whale blood transcriptome. The Trinity transcriptome assembly from beluga was comprised of 91,325 transcripts that represented a wide array of cellular functions and processes and was extremely similar in content to the blood transcriptome of another cetacean, the bottlenose dolphin. Expression of hemoglobin transcripts was much lower in beluga (25.6% of TPM, transcripts per million) than has been observed in many other mammals. A T12A amino acid substitution in the HBB sequence of beluga whales, but not bottlenose dolphins, was identified and may play a role in low temperature adaptation. The beluga blood transcriptome was extremely stable between sex and year, with no apparent clustering of samples by principle components analysis and <4% of genes differentially expressed (EBseq, FDR<0.05). While the impacts of season, sexual maturity, disease, and geography on the beluga blood transcriptome must be established, the presence of transcripts involved in stress, detoxification, and immune functions indicate that blood gene expression analyses may provide information on health status and exposure. This study provides a wealth of transcriptomic data on beluga whales and provides a sizeable pool of preliminary data for comparison with other studies in beluga whale.

WHOLE BLOOD TRANSFUSIONS TO TREAT SEVERE ANEMIA IN SEVEN COWNOSE RAYS ( RHINOPTERA BONASUS) AND ONE SHORT-TAIL STINGRAY ( DASYATIS BREVICAUDATA).

Blood transfusions can provide life-saving treatment to severely anemic animals. Due to limited availability and the difficulty of storing whole blood and blood products, such as fresh frozen plasma and packed red blood cells, exotic animals often receive fresh whole blood transfusions. Little is known about elasmobranch blood types and transfusions. Conspecific cross-matches within several different elasmobranch species were negative, indicating that in an emergency situation a single whole blood transfusion may be possible without causing a transfusion reaction. Experimental transfusions between healthy conspecific Atlantic rays ( Dasyatis sabina) showed no adverse reactions and autotransfusions in marbled electric rays ( Torpedo marmorata) were successful. There are no published reports of blood transfusions performed on clinically abnormal elasmobranchs. The following case series documents blood transfusions performed on seven cownose rays ( Rhinoptera bonasus) and one short-tail stingray ( Dasyatis brevicaudata). All rays were treated with the same protocol, which included pretreatment with steroids and antibiotics followed by an intravenous transfusion of freshly collected, heparinized, whole blood. Three animals survived and currently exhibit no abnormal clinical signs. Two animals died 55 days and 100 days post transfusion. Three animals died 2-22 days post transfusion. Although complications from blood transfusions could not be ruled out, all five animals that died had other health problems that likely contributed to their demise. All eight animals would almost certainly have died without a blood transfusion as they were severely anemic and moribund at the time of presentation. The methods described in this paper may be useful in the treatment of severely anemic elasmobranchs and this is the first report of blood transfusions in clinically abnormal elasmobranchs.