Ocular Examinations and Investigation of Intraocular Pressure, Tear Production, Central Corneal Thickness, and Corneal Touch Threshold in a Captive Flock of Atlantic Puffins (Fratercula arctica).

Ocular examinations were completed on a group of 10 Atlantic puffins (Fratercula arctica), 5 males and 5 females that ranged in age from 8 months to older than 30 years. The exams consisted of intraocular pressure/rebound tonometry, tear production/phenol red thread test, central corneal thickness/ultrasound pachymetry, and corneal sensitivity/esthesiometry. On ocular examination, there were no corneal abnormalities observed. Bilateral cataracts were diagnosed in 8 puffins, 6 of which were considered incipient, focal subcapsular opacities. One bird had hypermature cataracts and was removed from the study and excluded from data analysis; the other birds had no evidence of ophthalmic pathology that would interfere with diagnostic results (n = 9). All results for 9 birds were included in the study, with the exception of 1 puffin's tear production, which was too low for accurate assessment and was excluded from data analysis. There were no significant differences between right and left eye measurements for intraocular pressure, corneal thickness, and corneal sensitivity. The median intraocular pressure for both eyes (OU) was 13 mm Hg with an interquartile range [IQR] of 12-15 mm Hg. The median corneal thickness OU was 241 µm, IQR 233-248 µm. The median corneal sensitivity OU was 1.13 cm, IQR 0.81-1.50 cm. There was a significant difference between right and left eye measurements for tear production (right eye median, 7.5 mm/15 s, IQR 6.5-9.3 mm/15 s; and left eye median, 5.0 mm/15 s, IQR 4.0-7.3 mm/15 s) (P= .03), with the right eye producing more tears than the left. However, 1 puffin was determined to be an outlier, and when removed, there was no longer a significant difference (OU median, 7.0 mm/15 s, IQR 4.6-8.0 mm/15 s) (P = .38). There was no significant difference between sex and intraocular pressure, tear production, and corneal sensitivity. However, there was a significant difference between sex and corneal thickness (P = .02), with males (left eye median, 249 µm, IQR 241-249 µm) having thicker corneas than females (left eye median, 236 µm, 234-238 µm). Although sample size precluded statistical testing, there appeared to be an association between opacities and increasing age. There were no associations between age and intraocular pressure, tear production, or corneal thickness. There was a moderate correlation between age and corneal sensitivity, with older birds showing decreased corneal sensitivity (r = -0.57). Although the sample size of 9 birds was small, these findings provide preliminary ranges for ocular parameters of Atlantic puffins.

RISK ANALYSIS: CHANGING THE QUARANTINE PARADIGM?

As modern zoos evolved from menageries, quarantine practices were developed to protect animal collections from the introduction of diseases that might arrive with new animals from wild populations and, more recently, from other managed populations. As the understanding of, and ability to diagnose, diseases grew tremendously in recent decades, quarantine protocols became more stringent and standardized as the tolerance for disease introduction decreased. These practices resulted in healthier animal collections, and the introduction of transmissible pathogens of concern became rarer. Sustainable population management in zoos has further lessened the reliance on taking animals from the wild, and the knowledge of disease risk in such situations has deepened. Today animals being transferred between modern zoological institutions have well-documented medical histories complemented with robust collection health histories. This improved understanding has reduced concerns about disease transmission in many cases. Consequently, more formal use of risk analysis to drive transfer-related examination, diagnostic testing, and quarantine decision making was adopted by a number of zoos and aquariums that have modified their quarantine practices and use risk analysis to reduce quarantine requirements, in some cases moving animals directly into collections after transportation. This commentary discusses the rationale for and changes to quarantine processes for animal acquisitions in zoos and aquariums, using risk analysis to direct decision making.

COMPARISON OF A SMARTPHONE-BASED ELECTROCARDIOGRAM DEVICE WITH A STANDARD SIX-LEAD ELECTROCARDIOGRAM IN THE ATLANTIC BOTTLENOSE DOLPHIN ( TURSIOPS TRUNCATUS).

A bipolar, single-lead electrocardiogram (ECG) device is capable of recording ECGs with an integrated smartphone application. To determine the utility of this device, phone-based ECGs (pECG) were compared with standard six-lead ECGs (sECG) in four female Atlantic bottlenose dolphins ( Tursiops truncatus) at the National Aquarium. Study animals were trained to haul out onto a dry deck in ventral recumbency and allow simultaneous 30-sec ECG acquisition using the two devices. The pECG device was held against the thoracic wall caudal to the left axilla. The sECGs were recorded in the frontal plane. Instantaneous heart rates were obtained from identical QRS complexes on both ECGs. Three boarded cardiologists independently evaluated the rhythm and the polarity of the QRS depolarization for each recording and the results were compared. The mean heart rate was 80 beats/min (range 62-92 beats/min) and 80 beats/min (range 60-92 beats/min) for the pECG and sECGs, respectively. All four dolphins displayed sinus respiratory arrhythmia, and one animal had occasional atrial premature contractions. Rhythm diagnosis and QRS polarity were identical for the pECG and sECG. Dolphin vocalizations created artifacts on the pECG that were not present on the sECG, so pECGs had to be acquired without vocalization. The pECG appears to be an accurate and useful method to monitor the heart rate in Atlantic bottlenose dolphins. This system is inexpensive and portable, making it valuable for health examinations, transport monitoring, and stranding responses.

RETROSPECTIVE REVIEW OF MORTALITY IN GIANT PACIFIC OCTOPUS (ENTEROCTOPUS DOFLEINI).

The giant Pacific octopus (Enteroctopus dofleini) is a popular exhibit species in public display aquaria, but information on health and disease is limited. This retrospective review evaluates time in collection and describes antemortem clinical signs and pathology of giant Pacific octopuses in an aquarium setting. Between March 2004 and December 2013, there were 19 mortalities: eight males, 10 females, and one individual whose sex was not recorded. Average time spent in collection for all octopuses was 375 ± 173 days (males 351 ± 148 days, females 410 ± 196 days). Ten (52.6%) of the octopuses were sexually mature at the time of death, six (31.6%) were not sexually mature, and reproductive status could not be determined in three octopuses (15.8%). Minimal changes were noted on gross necropsy but branchitis was histologically evident in 14 octopuses, often in conjunction with amoeboid or flagellate parasites. Senescence, parasitism, and husbandry were all important contributors to mortality and should be considered when caring for captive octopuses.

Hydrocoelom and lymphedema in dendrobatid frogs at National Aquarium, Baltimore: 2003-2011.

Hydrocoelom and lymphedema are common in amphibians. In the family Dendrobatidae at the National Aquarium, Baltimore, prevalence was 3.19% from 2005 to 2011. Complete review of all cases in dendrobatids from 01 January 2003 to 01 June 2011 at the National Aquarium identified annual incidence rate, associated signs, diagnostic findings, therapeutics, and outcomes using medical record review. Associated etiologies were assessed via available necropsy and histopathology reports. Hydrocoelom was defined as coelomic distention with fluid; lymphedema was defined as fluid accumulation in the lymph spaces. These two syndromes could occur concurrently, and no association between case success (36.5% overall) or etiology was found to contrast hydrocoelom and lymphedema. Species susceptibility varied, with increased prevalence and incidence rate in Adelphobates castaneoticus, Dendrobates auratus, and Phyllobates spp. D. auratus also had reduced case success overall (26.0%). Females experienced hydrocoelom and/or lymphedema at an attack rate more than three times that of males. Fluid aspiration for culture was the most predictive antemortem diagnostic to find infectious etiologies, but antemortem diagnostics were largely noncontributory in identifying other causes of fluid accumulation. Enrofloxacin treatment increased case success more than fivefold when compared to those not treated and was the only treatment positively associated with case success. Infectious etiology, renal disease, and gastrointestinal disease, including parasitism, were seen commonly on postmortem evaluation of cases. These findings underscore the importance of the ability to individually track and treat cases, with fluid culture and initiation of enrofloxacin therapy (10 mg/kg p.o.q. 24 hr) seen as the best first steps after presentation with hydrocoelom or lymphedema.

KUDOA SP. (Myxozoa: Multivalvulida) in skeletal muscle of captive bullnose eagle rays, Myliobatis freminvillei (Rajiformes: Myliobatidae).

Kudoa spp. (Myxozoa: Multivalvulida) are of significant concern in marine and estuarine teleosts because of trophozoites and spores in the skeletal muscle reducing the marketability of wild and cultured fish. Kudoa spp. have rarely been reported from elasmobranchs. Spores of a Kudoa sp. were identified in two captive, mature bullnose eagle rays (Myliobatisfreminvillei) (Rajiformes: Myliobatidae). The male bullnose eagle ray (case 1) showed intractable weight loss, muscle wasting, and hyperkalemia, and was euthanized. The skeletal muscle showed myofiber degeneration, vacuolation, and necrosis, and spores of a Kudoa sp. were present multifocally through the muscle. The female bullnose eagle ray (case 2) had a severe, rapidly progressive, erosive lesion extending through the pectoral fin and was euthanized. Spores of a Kudoa sp. were not found in the lesion but were present in two disparate sections of skeletal muscle. There was no inflammation or degeneration associated with the infected myofibers in case 2. From case 1, morphometrics of 15 spores in histologic section were apical width 6.5-8.0 µm, apical thickness 5.0-7.0 µm, lateral width 5.5-6.5 µm, length 4.5-6.5 µm; the equal-sized polar capsules were spherical, 1.5-2.5 µm in diameter; there was a clear halo 2 µm thick around the spores, considered to be a mucous envelope. Spores from case 2 were indistinguishable from those in case 1. This appears to be the first report of Kudoa sp. in bullnose eagle rays.

Proliferative thyroid lesions in three diplodactylid geckos: Nephrurus amyae, Nephrurus levis, and Oedura marmorata.

Over a 5-mo period, three diplodactylid geckos housed at the National Aquarium were diagnosed with proliferative thyroid lesions: a rough knob-tail gecko (Nephrurus amyae), a smooth knob-tail gecko (Nephrurus levis), and a marbled velvet gecko (Oedura marmorata). Clinical signs included an intraoral mass or ventral throat swelling (or both), oral bleeding, and weight loss. Both of the knob-tail geckos died. The histologic diagnosis for the rough knob-tail gecko was thyroid carcinoma with metastases to the liver and lungs, and thyroid carcinoma with no metastases was reported in the smooth knob-tail gecko. A thyroidectomy was performed on the marbled velvet gecko with a histologic diagnosis of adenomatous hyperplasia. Postoperative weight loss and bradycardia resolved following oral supplementation with levothyroxine. The animal is in normal health 10 mo post-surgery. Five other diplodactylid geckos in the collection remain unaffected, giving a 38% prevalence of proliferative thyroid lesions (3/8). The etiology remains undetermined. This is the first report of a cluster of proliferative thyroid lesions in geckos.

PREVALENCE OF MULTIPLE REPTILIAN PATHOGENS IN THE OROPHARYNGEAL MUCOSA, CLOACAL MUCOSA, AND BLOOD OF DIAMONDBACK TERRAPIN (MALACLEMYS TERRAPIN) POPULATIONS FROM MARYLAND AND GEORGIA, USA.

The diamondback terrapin (Malaclemys terrapin) is a coastal turtle with a range from Massachusetts to Texas and is the only exclusively brackish water turtle in North America. Two populations of wild terrapins from Maryland (n=55) and Georgia (n=7) were examined and tested for potential reptile pathogens. Whole blood and a mucosal (combined oropharyngeal and cloacal) swab from each animal were evaluated by quantitative PCR for 15 potential pathogens including frog virus 3, box turtle Mycoplasmopsis, Mycoplasma agassizii, Mycoplasma testudineum, Salmonella Enteritidis, Salmonella Typhimurium, Borrelia burgdorferi, Anaplasma phagocytophilum, tortoise intranuclear coccidia, testudinid alphaherpesvirus 2, terrapene herpesvirus 1, and terrapene adenovirus. Swabs were positive for a DNA segment 100% homologous to M. testudineum in both populations, with Maryland animals 87% (48 of 55) positive and Georgia animals 86% (6 of 7) positive. Although Mycoplasmopsis spp. are important respiratory pathogens for members of the order Testudines, none of the animals in the study showed any sign of upper respiratory disease. Our data suggest that M. testudineum may survive in non-Testudinidae turtles without causing clinical sigs of disease and suggesting appropriate precautions should be taken in facilities that house multiple species of turtles simultaneously.

Phaeohyphomycosis due to Exophiala in Aquarium-Housed Lumpfish (Cyclopterus lumpus): Clinical Diagnosis and Description.

Phaeohyphomycosis caused by Exophiala species represents an important disease of concern for farmed and aquarium-housed fish. The objective of this study was to summarize the clinical findings and diagnosis of Exophiala infections in aquarium-housed Cyclopterus lumpus. Clinical records and postmortem pathology reports were reviewed for 15 individuals from 5 public aquaria in the United States and Canada from 2007 to 2015. Fish most commonly presented with cutaneous ulcers and progressive clinical decline despite topical or systemic antifungal therapy. Antemortem fungal culture of cutaneous lesions resulted in colonial growth for 7/12 samples from 8 individuals. Amplification of the internal transcribed spacer region (ITS) of nuclear rDNA identified Exophiala angulospora or Exophiala aquamarina in four samples from three individuals. Postmortem histopathologic findings were consistent with phaeohyphomycosis, with lesions most commonly found in the integument (11/15), gill (9/15), or kidney (9/15) and evidence of fungal angioinvasion and dissemination. DNA extraction and subsequent ITS sequencing from formalin-fixed paraffin-embedded tissues of seven individuals identified E. angulospora, E. aquamarina, or Cyphellophora sp. in four individuals. Lesion description, distribution, and Exophiala spp. identifications were similar to those reported in farmed C. lumpus. Antemortem clinical and diagnostic findings of phaeohyphomycosis attributable to several species of Exophiala provide insight on the progression of Exophiala infections in lumpfish that may contribute to management of the species in public aquaria and under culture conditions.

Fish quarantine: current practices in public zoos and aquaria.

The primary goal of quarantine is to reduce the risk of introducing infectious diseases into established collections. Fish quarantine is inherently complex because of the variety of species, environmental requirements, and facilities. To examine current practices, questionnaires were submitted to 60 public zoos and aquaria, predominantly in North America. Questions reviewed system type (closed, flow-through), quarantine length, diagnostics, treatments, and cleaning and disinfection. Forty-two of the 60 institutions responded. Most institutions had separate quarantine protocols for freshwater teleosts, marine teleosts, and elasmobranchs. Ninety-five percent of institutions had a minimum quarantine period of 30 days or more. Sixty-four percent of institutions used isolated areas for some or all of their fish quarantine. Twenty-five percent had designated fish quarantine staff. All institutions used regular visual examinations to assess animal health. Fifty-four percent of the institutions carried out routine hands-on diagnostics on some fish; this was more common for elasmobranchs than teleosts. All institutions carried out necropsies on mortalities. Fifteen percent of institutions performed histopathology on almost all fresh mortalities; 54% percent performed histopathology on less than 10% of mortalities. Prophylactic treatments were common in closed systems, in particular, formalin immersion for teleosts, freshwater dips and copper sulfate immersion for marine teleosts, and praziquantel immersion for marine teleosts and elasmobranchs. Institutions using dips generally did so at the start or end of quarantine. Fenbendazole- and praziquantel-medicated foods were used commonly in teleosts, but dosages varied greatly. Cleaning and disinfection of systems and equipment increased in response to known pathogens. These results can be used to compare and discuss fish quarantine practices at display facilities in order to improve quarantine success.

Development of methodology and reference intervals for the analysis of the free-ranging Atlantic horseshoe crab Limulus polyphemus hemolymph.

BACKGROUND: The Atlantic horseshoe crab, Limulus polyphemus, is valuable to fishing and biomedical industries, for education in public aquaria, and of ecological importance because the eggs provide a critical food source for migratory shorebirds. While hematologic and biochemical analyses are fundamental tools for health assessment, reports are limited for the four extant species of horseshoe crabs; L polyphemus (East Coast North America), Carcinoscorpius rotundicauda (mangrove horseshoe crab Southeast Asia), Tachypleus gigas, and Tachypleus tridentatus (Southeast and East Asia). OBJECTIVES: This prospective study measured hemolymph values to determine RIs from free-ranging horseshoe crabs during the annual breeding aggregation in Delaware Bay, USA. METHODS: Near-shore animals were briefly manually restrained for hemolymph collection from the cardiac sinus. Hematologic samples (25 males, 25 females) were collected for hemocytometer counts using the Natt and Herrick method. A second set of 50 animals (25 males, 25 females) was sampled for biochemistry measurands, including copper concentration by spectrophotometer, osmolality by vapor pressure osmometer, and protein electrophoresis. RESULTS: Total hemocyte counts varied widely (RI 6600-44 300/µL). Electrolytes were consistent with osmoconformers living in seawater. There was a marked difference in sexes, with females showing much lower protein, glucose, and copper levels. Eleven females had multiple outliers, and all data were excluded from RI calculations. CONCLUSIONS: Further studies of this species outside of spawning season are needed. Expanding our knowledge of horseshoe crab health is important to decreasing morbidity and mortality in research applications and maximizing longevity and overall health in display settings.

Cross matching of blood in carcharhiniform, lamniform, and orectolobiform sharks.

The transfusion of whole blood in elasmobranchs could provide cardiovascular support following hemorrhage. Since donor and recipient compatibility is not known, a technique was established to allow cross matching of red blood cells and serum in sharks. Cross matching was carried out among 19 individuals from seven species: the nurse shark (Ginglymostoma cirratum), sandbar shark (Carcharhinus plumbeus), sandtiger shark (Carcharias taurus), white-spotted bamboo shark (Chiloscyllium plagiosum), brown-banded bamboo shark (Chiloscyllium punctatum), zebra shark (Stegostoma fasciatum), and spotted wobbegong (Orectolobus maculatus). Negative cross-matches showed no agglutination or hemolysis, suggesting that donor and recipient would be compatible. Cross-matches between conspecifics were all negative (sandbar, sandtiger, nurse, and white-spotted bamboo sharks). All cross-matches between sandbar and sandtiger sharks were also negative. Positive crossmatches consisted of agglutination or hemolysis of red blood cells, suggesting that the donor and recipient would be incompatible. Strong positive reactions occurred, for example, with red blood cells from sandtiger and sandbar sharks and serum from nurse sharks. Cross matching should be carried out in elasmobranchs prior to any blood transfusion.

Amphibian Renal Disease.

Amphibians are a remarkably diverse group of vertebrates with lifestyles ranging from fully aquatic to entirely terrestrial. Although some aspects of renal anatomy and physiology are similar among all amphibians, species differences in nitrogenous waste production and broad normal variation in plasma osmolality and composition make definitive antemortem diagnosis of renal disease challenging. Treatment is often empirical and aimed at addressing possible underlying infection, reducing abnormal fluid accumulation, and optimizing husbandry practices to support metabolic and fluid homeostasis. This article reviews amphibian renal anatomy and physiology, provides recommendations for diagnostic and therapeutic options, and discusses etiologies of renal disease.

Coliform Bacteria Monitoring in Fish Systems: Current Practices in Public Aquaria.

Public aquaria evaluate coliform indicator bacteria levels in fish systems, but the purpose of testing, testing methods, and management responses are not standardized, unlike with the coliform bacteria testing for marine mammal enclosures required by the U.S. Department of Agriculture. An online survey was sent to selected aquaria to document current testing and management practices in fish systems without marine mammals. The information collected included indicator bacteria species, the size and type of systems monitored, the primary purpose of testing, sampling frequency, test methods, the criteria for interpreting results, corrective actions, and management changes to limit human exposure. Of the 25 institutions to which surveys were sent, 19 (76%) responded. Fourteen reported testing for fecal indicator bacteria in fish systems. The most commonly tested indicator species were total (86%) and fecal (79%) coliform bacteria, which were detected by means of the membrane filtration method (64%). Multiple types and sizes of systems were tested, and the guidelines for testing and corrective actions were highly variable. Only three institutions performed additional tests to confirm the identification of indicator organisms. The results from this study can be used to compare bacterial monitoring practices and protocols in fish systems, as an aid to discussions relating to the accuracy and reliability of test results, and to help implement appropriate management responses. Received August 23, 2015; accepted December 29, 2015.

Osmoregulation in fish. Mechanisms and clinical implications.

Fish have developed remarkable mechanisms for coping with life in water. The salinity/osmolarity of aquatic habitats can be quite variable. Fish have evolved mechanisms for maintaining fluid and electrolyte homeostasis across a wide range of salinities. Marine teleosts, freshwater teleosts, and marine elasmobranchs all utilize different physiologic strategies for osmoregulation. Kidneys do play a role in osmoregulation, but overall, extrarenal mechanisms are equally if not more important sites for maintaining osmotic homeostasis. Extrarenal sites include the gill tissue, the alimentary tract, the rectal gland (elasmobranchs), and the urinary bladder. Through an understanding of osmoregulatory physiology in these three groups of fishes, the effects of injury and disease on fluid and electrolyte balance can be elucidated. Armed with this knowledge and the results of diagnostic testing, safe and effective fluid/electrolyte support can then be instituted.

Assessment of mercury and selenium concentrations in captive bottlenose dolphin's (Tursiops truncatus) diet fish, blood, and tissue.

Concentrations of total mercury (Hg) and selenium (Se) were determined in diet fish and whole blood and tissue samples from seven bottlenose dolphins (Tursiops truncatus) housed at the National Aquarium Baltimore (NAB). In addition, concentrations of monomethylmercury (CH(3)Hg(+)) were determined in diet fish and dolphins' tissue samples. The data were compared with the values found in wild populations to better understand how the dietary Hg and Se uptake rates affect the Hg and Se levels in dolphins. The diet fish total Hg concentrations ranged between 14 and 47 ng g(-1) and were markedly lower than for similar fish found in Florida, South Carolina, and other aquaria. CH(3)Hg(+) accounted for 85 to 91% of the total Hg found in diet fish. The diet fish Se concentrations ranged between 270 and 800 ng g(-1), indicating excess molar concentrations of Se over Hg. The Hg concentration range in the blood of NAB dolphins was 27-117 ng g(-1) and the concentrations were about one order of magnitude and several factors lower than the concentrations found in the blood of wild bottlenose dolphins in Florida and in South Carolina, respectively. The total Hg and CH(3)Hg(+) in tissue samples were also significantly lower than the reported values obtained from wild populations of bottlenose dolphins. The differences in the Hg concentrations in the dolphins' blood may be due to the different levels of Hg atmospheric deposition in the area where the dolphins' diet fish were found. The Se concentration range in the blood of NAB dolphins was 221-297 ng g(-1) which was two factors lower than the values found in wild populations. The lower Hg levels, as well as higher Se:Hg molar ratios in the blood of NAB dolphins, suggest that NAB dolphins may be less susceptible to the potential neurotoxicity from the CH(3)Hg(+) in their blood.