Ancient DNA analysis of the Japanese sea lion (Zalophus californianus japonicus Peters, 1866): preliminary results using mitochondrial control-region sequences

In this study, we successfully extracted ancient DNA from skeletal remains of the Japanese sea lion-a species that is practically extinct-from archaeological sites and determined a partial sequence of its mitochondrial DNA control region. A molecular phylogenetic tree constructed by the neighbor-joining (NJ) method showed that the sequences from Japanese sea lions clustered together, with a high bootstrap value, and that this cluster was closest to the California sea lion cluster. The distinctly divergent cluster of Japanese sea lions reflected the morphological classification of these animals as a distinct species of the genus Zalophus; however, proximity to the California sea lion cluster simultaneously implied conformation with the traditional classification of these animals as a subspecies of Zalophus californianus. The average amount of nucleotide substitution between the Japanese and California sea lions was 7.02%. The Japanese and California sea lions were estimated to have diverged 2.2 million years ago, i.e., in the late Pliocene Epoch. This is the first report on a genetic analysis of the Japanese sea lion.     

Hepatic carcinoma with spleen metastasis in a California sea lion from the Gulf of California.

A primary hepatic carcinoma with a neuroendocrine pattern was detected in an adult female California sea lion (Zalophus californianus) found dead on Granito Island in the Gulf of California (Mexico) in January 1996. At necropsy, several light yellow nodules of different sizes were observed on the entire surface of the liver and spleen. Microscopic examination of these nodules using routine haematoxylin-eosin stain, revealed cubic, polyhedral and pleomorphic cells with three to four bizarre mitotic figures per field (40X). An immunohistochemistry test revealed a positive reaction of indirect immunoperoxide to cytokeratin (CK2). This is the first known case of a primary hepatic carcinoma in free-ranging California sea lions from Mexican waters.     

Pathology and preliminary characterization of a parapoxvirus isolated from a California sea lion (Zalophus californianus)

Cutaneous pox-like lesions are a common complication in the rehabilitation of pinnipeds. However, the exact identity, taxonomy, and host range of pinniped parapoxviruses remain unknown. During a poxvirus outbreak in May 2003 in California sea lions (Zalophus californianus) at a marine mammal rehabilitation facility, multiple raised, firm, 1-3-cm skin nodules from the head, neck, and thorax of one sea lion weanling pup that spontaneously died were collected. Histologically, the nodules were characterized by inflammation and necrosis of the dermis and epidermis, acanthosis, and ballooning degeneration of the stratum spinosum. Large, coalescing eosinophilic cytoplasmic inclusions were observed in the ballooned cells. A parapoxvirus (sea lion poxvirus 1, SLPV-1) was isolated on early passage California sea lion kidney cells inoculated with a tissue homogenate of a skin nodule. The morphology of the virions on electron microscopy was consistent with that of parapoxviruses. Partial sequencing of the genomic region encoding the putative major virion envelope antigen p42K confirmed the assignment of the sea lion poxvirus to the genus Parapoxvirus. Although SLPV-1 is most closely related to the poxvirus of harbor seals of the European North Sea, it is significantly different from orf virus, bovine papular stomatitis virus, pseudocowpox virus and the parapoxvirus of New Zealand red deer.     

Evidence of injury caused by gas bubbles in a live marine mammal: barotrauma in a California sea lion Zalophus californianus

A yearling male California sea lion Zalophus californianus with hypermetric ataxia and bilateral negative menace reflexes was brought to The Marine Mammal Center, Sausalito, California, U.S.A., in late 2009 for medical assessment and treatment. The clinical signs were due to multiple gas bubbles within the cerebellum. These lesions were intraparenchymal, multifocal to coalescing, spherical to ovoid, and varied from 0.5 to 2.4 cm diameter. The gas composed 21.3% of the total cerebellum volume. Three rib fractures were also noted during diagnostic evaluation and were presumed to be associated with the gas bubbles in the brain. The progression of clinical signs and lesion appearance were monitored with magnetic resonance imaging, cognitive function testing and computed tomography. Gas filled voids in the cerebellum were filled with fluid on follow up images. Clinical signs resolved and the sea lion was released with a satellite tag attached. Post release the animal travelled approximately 75 km north and 80 km south of the release site and the tag recorded dives of over 150 m depth. The animal re-stranded 25 d following release and died of a subacute bronchopneumonia and pleuritis. This is the first instance of clinical injury due to gas bubble formation described in a living pinniped and the first sea lion with quantifiable cerebellar damage to take part in spatial learning and memory testing.     

Metastrongyloid nematode (Otostrongylus circumlitus) infection in a stranded California sea lion (Zalophus californianus)--a new host-parasite association

A stranded yearling male California sea lion was admitted to a rehabilitation center June 2003. On presentation, the sea lion was emaciated and had diarrhea and neutrophilia. Two weeks later, the animal became anorexic, blood and mucus were observed around the oral cavity, and corneal opacity was noted in the right eye. Hematology results at that time included leukocytosis consisting of neutrophilia with a left shift, anemia, and thrombocytopenia. Despite supportive care, the sea lion died. On post mortem examination, there were multiple areas of hemorrhage scattered throughout all lung lobes, and pulmonary blood vessels were occluded by fibrin thrombi. Nematodes identified as immature forms of Otostrongylus circumlitus were found in the right ventricle and pulmonary arteries. Histologic findings in the lungs included severe suppurative and necrotizing arteritis with vascular thrombosis, interstitial pneumonia, and large areas of pulmonary hemorrhage. This report of O. circumlitus infection in a California sea lion (Zalophus californianus) might indicate a potentially new host-parasite association.     

Shunt dynamics in experimental atrial septal defects.

Inorder to study the hemodynamic variables involving the magnitude, direction, and timing of phasic shunt flow, both the interatrial pressure gradient and blood flow along with other pertinent hemodynamic variables were measured instantaneously across a surgically created atrial septal defect (ASD) in seven awake dogs. Atrial and ventricular pacing and infusion of phenylephrine and isoproterenol were used to alter hemodynamic conditions. The wave form of phasic ASD flow was similar both in configuration and timing to the interatrial pressure gradient. During the cardiac cycle, both left-to-right (L-R) and right-to-left (R-L) shunting occurred: atrial contraction augmented L-R flow; the onset of ventricular contraction was associated with R-L flow; during the latter part of ventricular contraction, flow returned to L-R with the maximum L-R shunting occurring in early diastole. Tachycardia, infusion of phenylephrine and isoproterenol did not alter the phasic flow pattern. Both spontaneous and positive pressure respiration decreased net L-R shunting.     

Isolation of a retrovirus and a herpesvirus from a captive California sea lion

A non-oncogenic retrovirus was isolated from an explanted skin biopsy from a captive California sea lion (Zalophus californianus) with a history of recurring skin lesions. The morphology of the viral particles in electron photomicrographs was characteristic of a foamy virus, a retrovirus in the subfamily Spumavirinae. Viral cytopathic effects consistent with foamy virus infection were observed in subsequent explants of skin and lymph nodes and co-cultivated peripheral blood leukocytes. The sea lion with the persistent foamy virus infection later died from pericarditis caused by Pasteurella multocida. A herpesvirus was isolated from explants of lung.     

Meningoencephalitis in two stranded California sea lions (Zalophus californianus) caused by aberrant trematode migration

Meningoencephalitis caused by aberrant trematode migration is described in two California sea lions (Zalophus californianus) admitted to a rehabilitation hospital between May and August 2001. Both animals displayed seizure activity and were euthanized due to poor response to therapy. Gross abnormal findings included liver flukes (Zalophotrema hepaticum) in the bile ducts and areas of swelling and necrosis in the cerebrum, cerebellum, and brain stem. Histopathology revealed meningoencephalitis with necrosis, hemorrhage, and many trematode eggs within the brain. In one sea lion, an adult trematode was found on the surface of the cerebrum. These are believed to be the first reported cases of meningoencephalitis caused by aberrant trematode migration in pinnipeds.     

Two distinct carcinomas of mammary gland origin in a California sea lion

Two histologic types of mammary cancer were encountered in an aged captive California sea lion (Zalophus californianus). A cancer with myoepithelial cell proliferation, which had metastasized to distant viscera, was located in the left cranial mammary region. Another cancer without myoepithelial cell proliferation was located in the right posterior mammary region, formed secondary nodules, and had metastasized to a regional lymph node. The presence of two different neoplasms in this sea lion is unusual.     

Isolation of a novel adenovirus from California sea lions Zalophus californianus

Viral hepatitis associated with adenoviral infection has been reported in California sea lions Zalophus californianus admitted to rehabilitation centers along the California coast since the 1970s. Canine adenovirus 1 (CAdV-1) causes viral hepatitis in dogs and infects a number of wildlife species. Attempts to isolate the virus from previous sea lion hepatitis cases were unsuccessful, but as the hepatitis had morphologic features resembling canine infectious hepatitis, and since the virus has a wide host range, it was thought that perhaps the etiologic agent was CAdV-1. Here, we identify a novel adenovirus in 2 stranded California sea lions and associate the infection with viral hepatitis and endothelial cell infection. Phylogenetic analysis confirmed the classification of the sea lion adenovirus in the Mastadenovirus genus with the most similarity to tree shrew adenovirus 1 (TSAdV-1, 77%). However, as the sea lion adenovirus appeared to be equally distant from the other Mastadenovirus species based on phylogenetic analysis, results indicate that it represents an independent lineage and species. Although sequences from this novel virus, otarine adenovirus 1 (OtAdV-1), show some similarity to CAdV-1 and 2, it is clearly distinct and likely the cause of the viral hepatitis in the stranded California sea lions.     

A bald encounter: hairless southern sea lion at the Falkland Islands

A hairless adult male southern sea lion (Otaria flavescens) was sighted at Sea Lion Island, Falkland Islands, on the 14th of August 2009. The sea lion was observed on several further occasions and was defending a small harem consisting of four females. To our knowledge, this is the first report of an apparently healthy sea lion being entirely hairless with the exception of vibrissae on the muzzle.     

The fecal viral flora of California sea lions

California sea lions are one of the major marine mammal species along the Pacific coast of North America. Sea lions are susceptible to a wide variety of viruses, some of which can be transmitted to or from terrestrial mammals. Using an unbiased viral metagenomic approach, we surveyed the fecal virome in California sea lions of different ages and health statuses. Averages of 1.6 and 2.5 distinct mammalian viral species were shed by pups and juvenile sea lions, respectively. Previously undescribed mammalian viruses from four RNA virus families (Astroviridae, Picornaviridae, Caliciviridae, and Reoviridae) and one DNA virus family (Parvoviridae) were characterized. The first complete or partial genomes of sapeloviruses, sapoviruses, noroviruses, and bocavirus in marine mammals are reported. Astroviruses and bocaviruses showed the highest prevalence and abundance in California sea lion feces. The diversity of bacteriophages was higher in unweaned sea lion pups than in juveniles and animals in rehabilitation, where the phage community consisted largely of phages related to the family Microviridae. This study increases our understanding of the viral diversity in marine mammals, highlights the high rate of enteric viral infections in these highly social carnivores, and may be used as a baseline viral survey for comparison with samples from California sea lions during unexplained disease outbreaks.     

Identification of two novel coccidian species shed by California sea lions (Zalophus californianus)

Routine fecal examination revealed novel coccidian oocysts in asymptomatic California sea lions (Zalophus californianus) in a rehabilitation facility. Coccidian oocysts were observed in fecal samples collected from 15 of 410 California sea lions admitted to The Marine Mammal Center between April 2007 and October 2009. Phylogenetic analysis using the full ITS-1 region, partial small subunit 18S rDNA sequence, and the Apicomplexa rpoB region identified 2 distinct sequence clades, referred to as Coccidia A and Coccidia B, and placed them in the Sarcocystidae, grouped with the tissue-cyst-forming coccidia. Both sequence clades resolved as individual taxa at ITS-1 and rpoB and were most closely related to Neospora caninum. Coccidia A was identified in 11 and Coccidia B in 4 of 12 sea lion oocyst samples successfully sequenced (3 of those sea lions were co-infected with both parasites). Shedding of Coccidia A oocysts was not associated with age class, sex, or stranding location, but yearlings represented the majority of shedders (8/15). This is the first study to use molecular phylogenetics to identify and describe coccidian parasites shed by a marine mammal.     

Coccidioidomycosis in a California sea lion (Zalophus californianus)

Coccidioidomycosis in an adult male California sea lion (Zalophus californianus) is described. The animal was housed in a zoo in Tucson, Arizona, for approximately 5 years. This is believed to be the first reported case of coccidioidomycosis in a marine mammal.     

Common cancer in a wild animal: the California sea lion (Zalophus californianus) as an emerging model for carcinogenesis

Naturally occurring cancers in non-laboratory species have great potential in helping to decipher the often complex causes of neoplasia. Wild animal models could add substantially to our understanding of carcinogenesis, particularly of genetic and environmental interactions, but they are currently underutilized. Studying neoplasia in wild animals is difficult and especially challenging in marine mammals owing to their inaccessibility, lack of exposure history, and ethical, logistical and legal limits on experimentation. Despite this, California sea lions (Zalophus californianus) offer an opportunity to investigate risk factors for neoplasia development that have implications for terrestrial mammals and humans who share much of their environment and diet. A relatively accessible California sea lion population on the west coast of the USA has a high prevalence of urogenital carcinoma and is regularly sampled during veterinary care in wildlife rehabilitation centres. Collaborative studies have revealed that genotype, persistent organic pollutants and a herpesvirus are all associated with this cancer. This paper reviews research to date on the epidemiology and pathogenesis of urogenital carcinoma in this species, and presents the California sea lion as an important and currently underexploited wild animal model of carcinogenesis.     

Comparative assessment of hemodynamic parameters in ventricular septal defect, obtained by Doppler echocardiography and catheterization of cardiac cavities

The study was undertaken to assess hemodynamic parameters by Doppler echocardiography in patients with ventricular septal defect (VSD) and pulmonary hypertension. Seventy-two patients aged 5 months to 9 years (mean 2.5 years) who had isolated VSD were examined. The authors conclude that it is possible and necessary to assess hemodynamics in the lesser circulation by using Doppler echocardiography. The method permits monitoring the time course of changes in the right heart, which makes it possible to follow the natural history of disease without applying invasive studies.     

Characterizing habitat suitability for a central‐place forager in a dynamic marine environment

Characterizing habitat suitability for a marine predator requires an understanding of the environmental heterogeneity and variability over the range in which a population moves during a particular life cycle. Female California sea lions (Zalophus californianus) are central‐place foragers and are particularly constrained while provisioning their young. During this time, habitat selection is a function of prey availability and proximity to the rookery, which has important implications for reproductive and population success. We explore how lactating females may select habitat and respond to environmental variability over broad spatial and temporal scales within the California Current System. We combine near‐real‐time remotely sensed satellite oceanography, animal tracking data (n = 72) from November to February over multiple years (2003–2009) and Generalized Additive Mixed Models (GAMMs) to determine the probability of sea lion occurrence based on environmental covariates. Results indicate that sea lion presence is associated with cool ( <14°C ), productive waters, shallow depths, increased eddy activity, and positive sea‐level anomalies. Predictive habitat maps generated from these biophysical associations suggest winter foraging areas are spatially consistent in the nearshore and offshore environments, except during the 2004–2005 winter, which coincided with an El Niño event. Here, we show how a species distribution model can provide broadscale information on the distribution of female California sea lions during an important life history stage and its implications for population dynamics and spatial management.     

First isolation of a calicivirus from the Steller sea lion (Eumetopias jubatus)

A calicivirus was isolated from the rectum of a Steller sea lion (Eumetopias jubatus) pup on Rogue Reef, off the southern Oregon coast. Based on the results of neutralization tests with specific typing antisera, the isolate was identified as San Miguel sea lion virus serotype 6 (SMSV-6). Blood obtained from nine of 37 pups (24%) during virus sample collection procedures had specific neutralizing antibodies to SMSV-6. The isolation of SMSV-6 from a Steller sea lion represents, to our knowledge, the first isolation of any virus from this widely distributed marine mammal species, and serves to reconfirm the host-nonspecificity of yet another calicivirus of marine origin.     

Production of chitinase and β-N-acetylglucosaminidase by intestinal bacteria of Pinnipedian animals

The chitinase- and β-N-acetylglucosaminidase(GlcNAcase)-producing ability of intestinal bacteria from Pinnipedian animals was determined using fluorogenic 4-methylumbelliferone glycosides of N-acetylglucosamine oligosaccharides. Intestinal microflora of a single Cape fur seal, three California sea lions and three South American sea lions were characterized by a predominance of isolates of the Bacteroidaceae and Enterobacteriaceae families and the genus Clostridium . Of the 711 isolates tested 26.0, 10.0 and 8.7% could hydrolyse 4-MU(GlcNAc)₁ 4-MU(GlcNAc)₂, and 4-MU(GlcNAc)₃, respectively. This result suggests that β-GlcNAcase producers occur at a higher density than do chitinase producers. Moreover, β-GlcNAcase, and to a lesser degree, chitinase seem to be efficiently produced by facultative anaerobes in the Cape fur seal and the California sea lion, and by both facultative and obligate anaerobes in the South American sea lion. To our knowledge, this is the first paper to report that isolates of the family Bacteroidaceae and the genus Streptococcus produce chitinase and/or β-GlcNAcase.     

Scintigraphie myocardique et séquelles de myocardite : présentation d’un cas clinique

Acute and subacute myocarditis are well-defined pathological entities but it is often difficult to identify them because their clinical expression is variable and the diagnosis is histological showing myocardial inflammation associated with degeneration and/or necrosis. Often symptoms are similar to those of acute pericarditis with “chest-crushing” pain that mimics myocardial infarction and prompts practitioners to request angiography, especially when there are cardiovascular risk factors. We report the case of a 61-year-old patient with neither cardiac history nor cardiovascular risk factor who consulted for a long and self-limited atypical chest pain with normal clinical examination and electrocardiogram. Myocardial perfusion scintigraphy showed a non-reversible photopenic area suspected of being a nontransmural necrosis or an artifact. The discovery of inferolateral hypokinesis of left ventricle on echocardiography led to perform a coronary angiography which was finally normal. Cardiac MRI allowed to diagnose scars of a previous episode of myocarditis.