Computed Tomography of the Mandibles of a Stranded Offshore Killer Whale (Orcinus orca).

A mature, adult female, offshore killer whale (Orcinus orca) was stranded deceased in Portage Bay, Alaska, in October 2015. Full necropsy examination with histopathology was performed. Consistent with previous studies of offshore killer whales, and thought to be a result of their unique elasmobranch diet, all the teeth were significantly abraded and almost flush with the gingival margin. Age was estimated at 30-35 years based on annuli and growth arrest lines in a remaining tooth. The dentate portion of the mandibles were excised en bloc and frozen until imaging could be completed. Radiography and computed tomography revealed lesions consistent with severe abrasion, pulp exposure and evidence of endodontic and/or periodontal disease in nine of the 15 mandibular teeth present (60.0%). Only five (33.3%) teeth were suspected to have been vital at the time of death based on imaging. Lesions were more severe rostrally, with the caudal teeth less affected. Autolysis precluded gingival histopathology and no teeth were analyzed histologically. Necropsy examination revealed a likely multifactorial cause of death, with most significant lesions including the severe chronic periodontal/endodontic disease with abrasion, inanition and emaciation with possible cardiovascular disease. This case highlights the importance of imaging in evaluating periodontal and endodontic status, especially post mortem when other tissues are no longer available, and demonstrates that periodontal and endodontic disease occur naturally in this species and can be a significant cause of morbidity in mature free-ranging killer whales of the offshore ecotype.

Immune status and function in harbor seal pups during the course of rehabilitation.

Routine hematological and serum chemistry parameters are important tools for the evaluation of health and the treatment of marine mammals admitted to rehabilitation centers. The evaluation of phagocytosis, oxidative burst and immunoglobulin G (IgG), as markers of immune system function, and haptoglobin (Hp), as a stress marker, were evaluated alongside routine hematology and chemistry as potentially informative diagnostic tools for marine mammal health assessments. Blood samples from harbor seal pups (Phoca vitulina) admitted to (n=46), and released from (n=28), the Vancouver Aquarium's Marine Mammal Rescue Center (VAMMRC) were collected (1) to perform routine and novel functional approaches to evaluate the health of pups at admission; (2) to determine how these parameters changed during the rehabilitation process; and (3) to generate baseline values for reference purposes. Sodium was the only blood parameter which differed between seal pups that survived and those that died, with the surviving pups exhibiting higher levels on admission diagnostics. Positive correlations between total protein concentrations, IgG and Hp levels were observed with globulin concentrations of seal pups. Changes in serum chemistry values between admission and release included a decrease in red blood cells (RBCs), glucose, bicarbonate, total bilirubin, γ-glutamyltransferase (GGT) levels, and an increase in mean cell volume (MCV), mean cell hemoglobin (MCH), mean cell hemoglobin concentration (MCHC), lymphocytes, eosinophils, urea, potassium, anion gap, calcium, phosphorus, total protein, albumin, globulin and osmolality levels. During the rehabilitation process, phagocytosis decreased, while Hp levels increased. Age and improved health appeared to underlie changes in these parameters during the rehabilitation period.

Mercury toxicity in beluga whale lymphocytes: limited effects of selenium protection.

Increasing emissions of anthropogenic mercury represents a growing concern to the health of high trophic level marine mammals. In its organic form, this metal bioaccumulates, and can be toxic to several physiological endpoints, including the immune system. In this study, we (1) evaluated the effects of inorganic mercury (mercuric chloride, HgCl2) and organic mercury (methylmercuric chloride, MeHgCl) on the in vitro function of lymphocytes isolated from the peripheral blood of beluga whales (Delphinapterus leucas); (2) characterized the potential protective effects of sodium selenite (Na2SeO3) on cell proliferation of HgCl2 or MeHgCl-treated beluga whale lymphocytes; and (3) compared these dose-dependent effects to measurements of blood Hg in samples collected from traditionally harvested beluga whales in the western Canadian Arctic. Lymphocyte proliferative responses were reduced following exposure to 1 µM of HgCl2 and 0.33 µM of MeHgCl. Decreased intracellular thiol levels were observed at 10 µM of HgCl2 and 0.33 µM of MeHgCl. Metallothionein induction was noted at 0.33 µM of MeHgCl. Concurrent exposure of Se provided a degree of protection against the highest concentrations of inorganic Hg (3.33 and 10 µM) or organic Hg (10 µM) for T-lymphocytes. This in vitro protection of Se against Hg toxicity to lymphocytes may contribute to the in vivo protection in beluga whales exposed to high Hg concentrations. Current Hg levels in free-ranging beluga whales from the Arctic fall into the range of exposures which elicited effects on lymphocytes in our study, highlighting the potential for effects on host resistance to disease. The implications of a changing Arctic climate on Hg fate in beluga food webs and the consequences for the health of beluga whales remain pressing research needs.

The effect of diet and feeding level on survival and weight gain of hand-raised harbor seal pups (Phoca vitulina).

Hundreds of stranded harbor seals pups (Phoca vitulina) are brought to wildlife rescue centers every year. Typical hand-rearing diets include artificial milk-replacers and diets based on macerated fish fed via gavage, but weight gains are often low and mortality rates can be high. This study compared survival and weight gain of orphaned seal pups fed either artificial milk-replacer or fish-formula. Pups admitted to the facility in summer 2007 (n=145) and 2008 (n=98) were randomly assigned to one of two diets and fed by gavage until weaning. In 2007, pups fed milk-replacer gained more (43 ± 12 g/d) than those fed fish-formula (loss of 13 ± 6 g/d; P<0.002). In 2008, when intake was increased from 8 to 11% of body weight daily, weight gain improved for both diets but remained higher in pups fed milk-replacer (123 ± 12 g/d, vs. loss of 6 ± 8 g/day; P<0.001). Pup survival to weaning was significantly higher in 2008 than 2007 (P<0.001) and was higher for pups on milk-replacer compared with those on fish-formula (P<0.05). Survival was also correlated with body weight at admittance (P<0.001). Although neither diet achieved the weight gains recorded in mother-raised pups (400-800 g/d), the artificial milk-replacer was clearly more successful, and pups fared better in the second year of the study when intake was higher.

Pathology and epidemiology of phocid herpesvirus-1 in wild and rehabilitating harbor seals (Phoca vitulina richardsi) in the northeastern Pacific.

Phocid herpesvirus-1 (PhHV-1, subfamily Alphaherpesvirinae) was isolated from harbor seals (Phoca vitulina vitulina) in the Netherlands in 1985, and was subsequently identified in Pacific harbor seals (Phoca vitulina richardsi) from California, USA in the 1990s. PhHV-1-associated pathology was first recognized in harbor seal carcasses submitted to a veterinary diagnostic laboratory in Abbotsford, British Columbia, Canada in 2000, and 63 cases were identified by 2008. A review of these cases indicated that PhHV-1-associated disease is widespread in harbor seals in the wild and within rehabilitation facilities in the coastal northeastern Pacific (including British Columbia, Canada, and Washington, USA). Morbidity and mortality occurred primarily in neonatal and weanling seal pups, and was due to PhHV-1 alone, or in combination with other disease processes. All cases occurred between July and October, corresponding to the pupping and weaning seasons in this area. Although previous publications have described the prevalence of antibody to PhHV-1 in harbor seals from British Columbia, Canada and Washington, USA this is the first study to focus on the epidemiology and pathology of the virus in this region.

Novel symptomatology and changing epidemiology of domoic acid toxicosis in California sea lions (Zalophus californianus): an increasing risk to marine mammal health.

Harmful algal blooms are increasing worldwide, including those of Pseudo-nitzschia spp. producing domoic acid off the California coast. This neurotoxin was first shown to cause mortality of marine mammals in 1998. A decade of monitoring California sea lion (Zalophus californianus) health since then has indicated that changes in the symptomatology and epidemiology of domoic acid toxicosis in this species are associated with the increase in toxigenic blooms. Two separate clinical syndromes now exist: acute domoic acid toxicosis as has been previously documented, and a second novel neurological syndrome characterized by epilepsy described here associated with chronic consequences of previous sub-lethal exposure to the toxin. This study indicates that domoic acid causes chronic damage to California sea lions and that these health effects are increasing.

Fetus in fetu in a harbor seal (Phoca vitulina richardi): histopathologic, genetic, and toxicologic analysis.

A young harbor seal (Phoca vitulina richardi), stranded on the coast of California, was found to have a 20-cm-diameter cranial cervical mass. Surgical excision revealed the subcutaneous mass to be covered in haired skin with multiple glabrous areas and structures resembling a jaw with tooth buds, eyelids, and a tail. The mass deformed the host pup's skull. Histologic examination revealed a complete vertebra in the tail, teeth in the jaw, and areas resembling tongue and larynx. Class 1 MHC sequences amplified by polymerase chain reaction from the mass and the host twin were identical. The mass was diagnosed as a fetus in fetu, a rare congenital anomaly in which 1 conjoined twin is completely enclosed in the body of the other twin. The host pup died, and no additional defects were found; however, blubber levels of persistent organic pollutants were high. The cause of the congenital anomaly in this pup is uncertain.

Pleomorphic rhabdomyosarcoma with pulmonary metastasis in a stranded Steller (northern) sea lion (Eumetopias jubatus).

A Steller (northern) sea lion (SSL), stranded in northern California in July 2000 had an anaplastic pleomorphic rhabdomyosarcoma in the latissimus dorsi muscle, with pulmonary metastasis. Diagnosis was based on light and electron microscopy and immunohistochemistry. Death was attributed to multiple parasitic and bacterial lesions. The SSL is of special concern because, for unknown reasons, the global population has declined by 50% over the last decade. Published post-mortem data, however, are scarce. This case report highlights several disease conditions that affect this species and is the first report of a malignant neoplasm in a free-ranging SSL.

Domoic acid toxicity in Californian sea lions (Zalophus californianus): clinical signs, treatment and survival.

Eighty-one Californian sea lions (Zalophus californianus) with signs of domoic acid toxicity stranded along the coast of California in 1998 when there were blooms of the domoic acid-producing alga Pseudonitzschia australis off-shore. In 2000, a further 184 sea lions stranded with similar clinical signs, but the strandings occurred both during detectable algal blooms and after the blooms had subsided. The clinical signs in these 265 Californian sea lions included seizures, ataxia, head weaving, decreased responsiveness to stimuli and scratching behaviour. Affected animals had high haematocrits, and eosinophil counts, and high activities of serum creatine kinase. They were treated supportively by using fluid therapy, diazepam, lorazepam and phenobarbitone. Fifty-five of the 81 sea lions (68 per cent) affected in 1998 and 81 of the 184 (44 per cent) affected in 2000 died despite the treatment. Three of the 23 sea lions which survived in 1998 were tracked with satellite and radiotransmitters; they travelled as far south as San Miguel Island, California, and survived for at least three months. Eleven of the 129 animals which were released stranded within four months of being released.

Use of medetomidine-zolazepam-tiletamine with and without atipamezole reversal to immobilize captive California sea lions.

From June 1998 to August 1999, 39 California sea lions (Zalophus californianus) were immobilized at a rehabilitation center in northern California (USA) using medetomidine plus zolazepam and tiletamine (MZT), alone and in combination with isoflurane, with atipamezole reversal. Animals were given 70 microg/kg medetomidine with 1 mg/kg of a 1:1 solution of tiletamine and zolazepam intramuscularly. Mean (+/-SD) time to maximal effect was 5+/-3 min. At the end of the procedure, animals were given 200 microg/kg atipamezole intramuscularly. Immobilization and recovery times were, respectively, 28+/-18 and 9+/-7 min for 15 animals maintained with MZT alone and 56+/-47 and 9+/-6 min for 18 animals intubated and maintained with isoflurane. One mortality occurred during anesthesia. Other disadvantages of the MZT combination included some prolonged ataxia, weakness and disorientation during recovery. However, the use of MZT resulted in faster induction and a more reliable plane of anesthesia that was reversible with atipamezole and safer than other previously used intramuscular agents. Physiological parameters including heart rate, respiratory rate, temperature, pulse oximeter saturation, and end-tidal carbon dioxide were monitored.

Humoral immune responses to phocine herpesvirus-1 in Pacific harbor seals (Phoca vitulina richardsii) during an outbreak of clinical disease.

Infection with phocine herpesvirus type-1 (PHV-1) has been associated with morbidity and high mortality in neonatal harbor seals (Phoca vitulina). A PHV-1 specific indirect enzyme linked immunosorbent assay (ELISA) was developed to sequentially measure the serological status of 106 harbor seal neonates admitted to a Pacific coast rehabilitation center (total number of sera tested was 371). Early in the season (February-April), the majority of pups had low serum levels of PHV-1 specific antibody. A dramatic increase in PHV-1 specific antibody, involving the majority of hospitalized pups, was observed during a 4-week period in May. This coincided with a high incidence of PHV-1 associated adrenal lesions and mortality. Although there was overall agreement between the timing of seroconversion to PHV-1 and histological evidence of PHV-1 infection, 82.4% of individual pups with adrenalitis had no evidence of a humoral response to PHV-1 at the time of their death. This suggests either a rapid disease course, or an inability to develop a humoral response in some neonatal seals.

Immobilization of California sea lions using medetomidine plus ketamine with and without isoflurane and reversal with atipamezole.

The use of medetomidine and ketamine, alone and in combination with isoflurane, with atipamezole reversal was evaluated for immobilizing 51 California sea lions (Zalophus californianus) for a variety of medical procedures at a rehabilitation center in northern California (USA) between May 1997 and August 1998. Animals were given 140 microg/kg medetomidine with 2.5 mg/kg ketamine intramuscularly. Mean (+/-SD) time to maximal effect was 8+/-5 min. At the end of the procedure, animals were given 200 microg/kg atipamezole intramuscularly. Immobilization and recovery times were, respectively, 25+/-12 and 9+/-7 min for 35 animals maintained with medetomidine and ketamine alone and 58+/-30 and 9+/-9 min for 16 animals intubated and maintained with isoflurane. No mortalities occurred as a result of the immobilizations. Disadvantages of the medetomidine and ketamine combination included a moderate variation in time to maximal effect and plane of sedation, a large injection volume and high cost. However, this combination offers safe and reversible immobilization that can be easily administered by the intramuscular route and that produces a plane of anesthesia that is sufficient to carry out most routine diagnostic procedures.

Mortality of sea lions along the central California coast linked to a toxic diatom bloom.

Over 400 California sea lions (Zalophus californianus) died and many others displayed signs of neurological dysfunction along the central California coast during May and June 1998. A bloom of Pseudo-nitzschia australis (diatom) was observed in the Monterey Bay region during the same period. This bloom was associated with production of domoic acid (DA), a neurotoxin that was also detected in planktivorous fish, including the northern anchovy (Engraulis mordax), and in sea lion body fluids. These and other concurrent observations demonstrate the trophic transfer of DA resulting in marine mammal mortality. In contrast to fish, blue mussels (Mytilus edulus) collected during the DA outbreak contained no DA or only trace amounts. Such findings reveal that monitoring of mussel toxicity alone does not necessarily provide adequate warning of DA entering the food web at levels sufficient to harm marine wildlife and perhaps humans.