Pamela A. Tuomi

ASSESSMENT OF ULTRASOUND IMAGING AS A NONINVASIVE MEASURE OF BLUBBER THICKNESS IN PINNIPEDS

Abstract Portable ultrasound imaging was examined as a noninvasive measure of skin including blubber thickness in captive subadult Steller sea lions (Eumetopias jubatus) and adult harbor seals (Phoca vitulina). This method was validated through comparison with blubber biopsy. Ultrasound images provided depth measurements that were accurate to 99.8% of the actual. This method allowed clear images of the epidermis, dermis, and blubber layer to be rapidly obtained, with minimal animal restraint, and allowed differentiation of phocid and otariid blubber structure.

Assessment of clinical pathology and pathogen exposure in sea otters (Enhydra lutris) bordering the threatened population in Alaska.

Northern sea otter (Enhydra lutris kenyoni) abundance has decreased dramatically over portions of southwest Alaska, USA, since the mid-1980s, and this stock is currently listed as threatened under the Endangered Species Act. In contrast, adjacent populations in south central Alaska, USA, and Russia have been stable to increasing during the same period. Sea otters bordering the area classified in the recent decline were live-captured during 2004–2006 at Bering Island, Russia, and the Kodiak Archipelago, Alaska, USA, to evaluate differences in general health and current exposure status to marine and terrestrial pathogens. Although body condition was lower in animals captured at Bering Island, Russia, than it was at Kodiak, USA, clinical pathology values did not reveal differences in general health between the two regions. Low prevalences of antibodies (>5%) were found in Kodiak, USA, and on Bering Island, Russia, to Toxoplasma gondii, Sarcocystis neurona, and Leptospira interrogans. Exposure to phocine herpesvirus-1 was found in both Kodiak, USA (15.2%), and Bering Island, Russia (2.3%). Antibodies to Brucella spp. were found in 28% of the otters tested on Bering Island, Russia, compared with only 2.7% of the samples from Kodiak, USA. Prevalence of exposure to Phocine distemper virus (PDV) was 41% in Kodiak, USA, but 0% on Bering Island, Russia. Archived sera from southwest and south-central Alaska dating back to 1989 were negative for PDV, indicating exposure occurred in sea otters in Kodiak, USA, in recent years. Because PDV can be highly pathogenic in naïve and susceptible marine mammal populations, tissues should be examined to explore the contribution of this virus to otter deaths. Our results reveal an increase in exposure to pathogens in sea otters in Kodiak, Alaska, USA, since the 1990s.

Intraperitoneal implantation of life-long telemetry transmitters in otariids

BackgroundPinnipeds, including many endangered and declining species, are inaccessible and difficult to monitor for extended periods using externally attached telemetry devices that are shed during the annual molt. Archival satellite transmitters were implanted intraperitoneally into four rehabilitated California sea lions (Zalophus californianus) and 15 wild juvenile Steller sea lions (Eumetopias jubatus) to determine the viability of this surgical technique for the deployment of long-term telemetry devices in otariids. The life history transmitters record information throughout the life of the host and transmit data to orbiting satellites after extrusion following death of the host.ResultsSurgeries were performed under isoflurane anesthesia and single (n = 4) or dual (n = 15) transmitters were inserted into the ventrocaudal abdominal cavity via an 8.5 to 12 cm incision along the ventral midline between the umbilicus and pubic symphysis or preputial opening. Surgeries lasted 90 minutes (SD = 8) for the 19 sea lions. All animals recovered well and were released into the wild after extended monitoring periods from 27 to 69 days at two captive animal facilities. Minimum post-implant survival was determined via post-release tracking using externally attached satellite transmitters or via opportunistic re-sighting for mean durations of 73.7 days (SE = 9.0, Z. californianus) and 223.6 days (SE = 71.5, E. jubatus).ConclusionThe low morbidity and zero mortality encountered during captive observation and post-release tracking periods confirm the viability of this surgical technique for the implantation of long-term telemetry devices in otariids.

Effects of ACTH, capture, and short term confinement on glucocorticoid concentrations in harlequin ducks (Histrionicus histrionicus)

Little is known about baseline concentrations of adrenal hormones and hormonal responses to stress in sea ducks, although significant population declines documented in several species suggest that sea ducks are exposed to increased levels of environmental stress. Such declines have been observed in geographically distinct harlequin duck populations. We performed an adrenocorticotropic hormone (ACTH) challenge to evaluate adrenal function and characterize corticosterone concentrations in captive harlequin ducks and investigated the effects of capture, surgery, and short term confinement on corticosterone concentrations in wild harlequin ducks. Harlequin ducks responded to the ACTH challenge with an average three-fold increase in serum corticosterone concentration approximately 90 min post injection, and a four- to five-fold increase in fecal glucocorticoid concentration 2 to 4 h post injection. Serum corticosterone concentrations in wild harlequin ducks increased within min of capture and elevated levels were found for several hours post capture, indicating that surgery and confinement maintain elevated corticosterone concentrations in this species. Mean corticosterone concentrations in wild harlequin ducks held in temporary captivity were similar to the maximum response levels during the ACTH challenge in captive birds. However, large variation among individuals was observed in responses of wild birds, and we found additional evidence suggesting that corticosterone responses varied between hatch year and after hatch year birds.

Chemical immobilization of Weddell seals (Leptonychotes weddellii) by ketamine/midazolam combination.

OBJECTIVE To provide reliable, effective immobilization for Weddell seals under extreme field conditions using an injectable ketamine/midazolam combination. STUDY DESIGN Observational study. ANIMALS Thirty adult Weddell seals (12 male, 18 female) in Erebus Bay, Antarctica, body mass (mean ± SD) 412 ± 47 kg, aged 9-27 years. METHODS Seals were immobilized with a target dose of 2 mg kg-1 ketamine hydrochloride and 0.1 mg kg-1 midazolam hydrochloride (IM), based on visually estimated body mass. When required, maintenance doses were administered at a target of 0.5 mg kg-1 ketamine hydrochloride and 0.025 mg kg-1 midazolam hydrochloride (IV). RESULTS Complete immobilization was achieved in 33 of 40 injections (14 of which were repeat events on the same individual). Time to immobilization averaged 12 ± 4 minutes, with a duration of initial immobility of 38 ± 19 minutes. Total immobilization time varied by handling protocol, including condition assessment and muscle biopsy (Protocol 1, 60 ± 13 minutes), condition assessment and instrument attachment (Protocol 2, 154 ± 13 minutes), and condition assessment, muscle biopsy and instrument retrieval (Protocol 3, 48 ± 8 minutes). Overall, a total immobilization time of 114 ± 60 minutes was accomplished with 4 ± 4 maintenance doses, and an average recovery time of 36 ± 17 minutes. Most effects of the anesthetic combination were unrelated to mass, age, sex or total body fat. However, leaner seals had longer duration of initial immobility (% and kg total body fat) and recovery times (kg fat). Apnea events were uncommon and treated effectively with doxapram. No animals died. CONCLUSIONS AND CLINICAL RELEVANCE Reliable and effective field immobilization of Weddell seals was accomplished with a low dose of ketamine hydrochloride and midazolam hydrochloride, utilizing IM injection initially and IV maintenance methods.OBJECTIVE To provide reliable, effective immobilization for Weddell seals under extreme field conditions using an injectable ketamine/midazolam combination. STUDY DESIGN Observational study. ANIMALS Thirty adult Weddell seals (12 male, 18 female) in Erebus Bay, Antarctica, body mass (mean +/- SD) 412 +/- 47 kg, aged 9-27 years. METHODS Seals were immobilized with a target dose of 2 mg kg(-1) ketamine hydrochloride and 0.1 mg kg(-1) midazolam hydrochloride (IM), based on visually estimated body mass. When required, maintenance doses were administered at a target of 0.5 mg kg(-1) ketamine hydrochloride and 0.025 mg kg(-1) midazolam hydrochloride (IV). RESULTS Complete immobilization was achieved in 33 of 40 injections (14 of which were repeat events on the same individual). Time to immobilization averaged 12 +/- 4 minutes, with a duration of initial immobility of 38 +/- 19 minutes. Total immobilization time varied by handling protocol, including condition assessment and muscle biopsy (Protocol 1, 60 +/- 13 minutes), condition assessment and instrument attachment (Protocol 2, 154 +/- 13 minutes), and condition assessment, muscle biopsy and instrument retrieval (Protocol 3, 48 +/- 8 minutes). Overall, a total immobilization time of 114 +/- 60 minutes was accomplished with 4 +/- 4 maintenance doses, and an average recovery time of 36 +/- 17 minutes. Most effects of the anesthetic combination were unrelated to mass, age, sex or total body fat. However, leaner seals had longer duration of initial immobility (% and kg total body fat) and recovery times (kg fat). Apnea events were uncommon and treated effectively with doxapram. No animals died. CONCLUSIONS AND CLINICAL RELEVANCE Reliable and effective field immobilization of Weddell seals was accomplished with a low dose of ketamine hydrochloride and midazolam hydrochloride, utilizing IM injection initially and IV maintenance methods.

Genotypic characterization of Streptococcus infantarius subsp. coli isolates from sea otters with infective endocarditis and/or septicemia and from environmental mussel samples

ABSTRACT Pulsed-field gel electrophoresis (PFGE) was used to type 128 Streptococcus infantarius subsp. coli isolates from sea otters and mussels. Six SmaI PFGE groups were detected, with one predominant group representing 57% of the isolates collected over a wide geographic region. Several sea otter and mussel isolates were highly related, suggesting that an environmental infection source is possible.

NOVEL POXVIRUS INFECTION IN NORTHERN AND SOUTHERN SEA OTTERS (ENHYDRA LUTRIS KENYONI AND ENHYDRA LUTRIS NEIRIS), ALASKA AND CALIFORNIA, USA

Abstract Small superficially ulcerated skin lesions were observed between October 2009 and September 2011 during captive care of two orphaned sea otter pups: one northern (Enhydra lutris kenyoni) in Alaska and one southern (Enhydra lutris nereis) in California. Inclusions consistent with poxviral infection were diagnosed by histopathology in both cases. Virions consistent with poxvirus virions were seen on electron microscopy in the northern sea otter, and the virus was successfully propagated in cell culture. DNA extraction, pan-chordopoxviral PCR amplification, and sequencing of the DNA-dependent DNA polymerase gene revealed that both cases were caused by a novel AT-rich poxvirus. Bayesian and maximum likelihood phylogenetic analyses found that the virus is divergent from other known poxviruses at a level consistent with a novel genus. These cases were self-limiting and did not appear to be associated with systemic illness. To our knowledge, this is the first report of a poxvirus in a mustelid species. The source of this virus, mode of transmission, zoonotic potential, and biological significance are undetermined.

Vitamin a deficiency and hepatic retinol levels in sea otters, Enhydra lutris

Abstract Vitamin A deficiency has rarely been reported in captive or free-ranging wildlife species. Necropsy findings in two captively housed southern sea otters (Enhydra lutris nereis) included irregular thickening of the calvaria characterized by diffuse hyperostoses on the internal surface. One animal also had moderate squamous metaplasia of the seromucinous glands of the nose. There was no measurable retinol in the liver of either sea otter. For comparison, hepatic retinol concentration was determined for 23 deceased free-ranging southern and northern (Enhydra lutris kenyoni) sea otters from California and Alaska. Free-ranging otters were found to have similar hepatic retinol concentrations (316 ± 245 mg/kg wet weight) regardless of their location and subspecies. All of these values were significantly higher than the levels in the affected animals. Consumption of a diet with very low vitamin A concentrations and noncompliance in daily supplementation are hypothesized as the causes of vitamin A deficiency in these two sea otters.

Description of a hair follicle mite (Acari: Demodecidae) parasitic in the earless seal family Phocidae (Mammalia: Carnivora) from the harbor seal Phoca vitulina Linnaeus, 1758

Abstract Demodex phocidi n. sp. is described (all life stages) from a captive Atlantic harbor seal, Phoca vitulina, from Seward, Alaska. This is the first description of a demodecid hair follicle mite from the earless seal family Phocidae. The mites reside in the sebaceous glands of the hair follicles and are associated with persistent areas of hyperkeratosis.

Best practice recommendations for the use of fully implanted telemetry devices in pinnipeds

Electronic telemetry devices have enabled many novel and important data collection and experimental opportunities for difficult to observe species. Externally attached devices have limited retention and may affect thermoregulation, energetics, social and reproductive behavior, visibility, predation risk and entanglement. Internally placed, surgically implanted devices can mitigate some of these effects and may open additional experimental opportunities. However, improper implementation can significantly affect animals and data. From a review of recent studies using fully implanted tags and studying their effects, we present 15 specific best practice recommendations for the use of such tags in pinnipeds. Recommendations address issues including device size, coating and sterilization, implantation surgery and effect assessment, within the framework of the Three R’s: Reduction, Refinement, Replacement. While developed for pinnipeds, these recommendations could apply to other aquatic mammals and vertebrates and to partially implanted or even external tags.