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Mass mortality in seals caused by a newly discovered morbillivirus.

During a recent disease outbreak among harbour seals (Phoca vitulina) in the North and Baltic seas, more than 17,000 animals have died. The clinical symptoms and pathological findings were similar to those of distemper in dogs. Based on a seroepizootiological study, using a canine distemper virus (CDV) neutralization assay, it was shown that CDV or a closely related morbillivirus (phocid distemper virus-PDV) was the primary cause of the disease. The virus was isolated in cell culture from the organs of dead seals and characterized as a morbillivirus by serology (immunofluorescence neutralization and enzyme-linked immunosorbent assays) and by negative contrast electron microscopy. Experimental infection of SPF dogs resulted in the development of mild clinical signs of distemper and CDV-neutralizing antibodies. The disease was reproduced in seals by experimental inoculation of organ material from animals that had died during the outbreak. However, seals that had been vaccinated with experimental inactivated CDV vaccines were protected against this challenge. This fulfilled the last of Koch's postulates, confirming that the morbillivirus isolated from the seal organs, was the primary cause of the disease outbreak. The recent demonstration of the presence of a similar virus in Lake Baikal seals (Phoca sibirica), which infected these Siberian seals 1 year before the northwestern European seals were infected, raises new questions about the origin of this infectious disease in pinnipeds.

Animals↗

Comparison of polychlorinated biphenyl (PCB) induced effects on innate immune functions in harbour and grey seals.

Polychlorinated biphenyls (PCBs) are known to have detrimental effects on the innate immune system of several mammalian species. Top predators such as marine mammals may be badly affected as PCBs can bioaccumulate in their blubber to high concentrations and previous studies have suggested that harbour seals may be particularly vulnerable to the immunotoxic effects of such contaminants. To investigate the effects of PCBs on innate immune functions in phocid seals, blood samples were collected from harbour and grey seals and exposed in vitro to a mixture of Aroclors. Separated mononuclear (PBMCs) and polymorphonuclear (PMNCs) leukocytes from each species were incubated with Aroclors (at 3 and 30 ngml(-1)) for 3 and 24 h incubation periods, after which phagocytosis, respiratory burst and cytotoxic activity were measured. The phagocytic activity of harbour seal PMNCs was decreased at both incubation times and at both Aroclor concentrations tested, but there was no effect on the grey seals. Similarly, the respiratory burst activity of harbour seals was decreased at both incubation times, but only at the higher concentration used. There were no differences in the cytotoxic activity of the PBMCs with respect to incubation times or concentrations in either species. However, differences were observed in the level of cytotoxic activity against YAC-1 target cells, with the grey seal PBMCs showing higher levels of activity. The observed differences in phagocytosis, respiratory burst and cytotoxic activity of the leukocytes following incubation with PCBs may have implications for the previously recorded differences in disease susceptibility between grey and harbour seals.

Animals↗

Isolation and partial characterization of a parapoxvirus isolated from a skin lesion of a Weddell seal.

A solitary skin lesion was found on the neck of a Weddell seal (Leptonychotes weddellii), chemically immobilized in Queen Maud Land (70 degrees 09'S, 05 degrees 22'E) Antarctica 2001. The lesion was elevated and 3cm in diameter, consisting of partly fresh and partly necrotic tissue, and proliferative papilloma-like structures were seen. Electron microscopy on a biopsy from the lesion revealed typical parapoxvirus particles. Polymerase chain reaction (PCR; B2L gene) generated amplicons of approximately 594 base pairs, comparable to Orf-virus, the prototype parapoxvirus. A comparison of these B2L PCR amplicon DNA sequences with corresponding sequences from other parapoxviruses, showed that the Weddell seal virus resembled isolates from grey seal (Halichoerus grypus) and harbour seal (Phoca vitulina) more than parapoxvirus from red deer (Cervus elaphus), sheep, cattle and Japanese serows (Capricornis crispus). It is thus concluded that the Weddell seal parapoxvirus belong to the tentative seal parapoxvirus species. Since parapox and orthopoxviruses may cause similar clinical diseases, we suggest that the term sealpox should be restricted to the clinical disease, whereas seal parapoxvirus should be used when caused by a parapoxvirus, rather than the general term "sealpox virus". This is the first verified case of parapoxvirus infection in a Weddell seal, and also the first report of any such infections in the Antarctic.

Animals↗

Serum leptin concentration during the terrestrial phase of the Southern elephant seal Mirounga leonina (Carnivora: Phocidae).

This study aimed to verify the serum leptin concentration during the terrestrial phase of the Southern elephant seal (Mirounga leonina) on Elephant Island (South Shetlands, Antarctica). Sera from 25 adult Southern elephant seals were analyzed. Leptin concentration was determined using the 125I radioimmunoassay method. Total protein, triglycerides, and cholesterol concentration were measured by spectrophotometric methods. Seals were grouped by sex and their physiological status (reproducing or molting seals). In reproducing seals, serum concentrations of leptin, total protein, triglycerides, and cholesterol were, respectively, 9.33+/-1.97 ng/ml, 6.87+/-0.09 g/dl, 98.26+/-2.12 mg/dl, and 232.17+/-41.18 mg/dl in males and 5.30+/-1.36 ng/ml, 6.44+/-0.29 g/dl, 109.01+/-3.34 mg/dl, and 219.20+/-26.65 in females (mean+/-1 SD). In molting seals, these values were 2.35+/-1.51 ng/ml, 7.42+/-0.25 mg/dl, 321.10+/-20.01 mg/dl, and 244.66+/-22.24 mg/dl in males and 2.94+/-1.89 ng/ml, 7.88+/-0.46 mg/dl, 197.54 mg/dl, and 224.55+/-16.70 mg/dl in females. In both males and females, there were no significant differences (P>0.05) in total protein and cholesterol concentration between reproducing and molting seals. However, both males and females showed higher leptin concentration (P<0.05) and lower triglyceride concentration (P<0.05) in the reproductive period than in the molt period. In the reproductive period, it was expected that the Southern elephant seal shows an inhibited hunger sensation, mobilizes stored energy, and stimulates the HPG axis. Results from the present study support this hypothesis and suggest that this strategy may be effected by the hormonal stimulation of leptin.

Adaptation, Physiological↗

Distribution of cytochrome P4501A (CYP1A) in the tissues of Baltic ringed and grey seals.

Information about the expression of CYP1A in wildlife species is essential for understanding the impact of organochlorine exposure on the health status of an exposed population. Therefore, we aimed at characterising a putative CYP1A enzyme expression in both hepatic and extrahepatic tissues of ringed and grey seals from the Baltic Sea and from less polluted waters. The cellular localisation of CYP1A was identified using a monoclonal antibody against scup P4501A1 (MAb 1-12-3). Immunohistochemical staining showed the highest level of CYP1A expression in liver hepatocytes, and the second highest level in the endothelial cells of capillaries and larger blood vessels in the liver and other organs. The most frequent and strongest staining was found in Baltic ringed seals. Although CYP1A-positive staining was observed in only a few tissues in the other seal populations, it was more intense in Baltic grey seals than in Canadian grey seals. The CYP1A enzyme activity, expressed as ethoxyresorufin O-deethylation (EROD), followed a similar tissue distribution and geographical pattern as the immunohistochemistry with clearly elevated EROD activities in most tissues of both Baltic seal populations. Immunochemical characterisation by immunoblotting confirmed the presence and elevation pattern of a putative CYP1A protein in ringed and grey seals, supporting our findings using other methods. The evenly distributed elevation of CYP1A expression among most of the tissues examined indicates that Baltic seals are exposed to CYP1A inducing agents affecting the whole body. This may result in an increased or decreased toxic potential of foreign substances, which may ultimately determine the biological effects of the contaminants.

Animals↗

cDNA cloning of an aryl hydrocarbon receptor from Baikal seals (Phoca sibirica).

Species differences in sensitivity to related planar halogenated aromatic hydrocarbons (PHAH) add significant uncertainty in assessing the ecological risk to aquatic mammals. To investigate mechanisms of PHAH sensitivity in aquatic mammals, we cloned and sequenced the cDNA of Baikal seal aryl hydrocarbon receptor (AHR), an intracellular protein that initiates PHAH-mediated effects. The Baikal seal AHR cDNA has an open reading frame of 843 amino acid residues with a predicted molecular mass of 94.6 kDa. Comparison of AHR amino acid sequences indicated a high degree of sequence conservation (98%) between Baikal and harbor seals. The high conservation of AHRs between Baikal and harbor seals indicates that these seals express AHR proteins closely related structurally. In our previous report (Kim & Hahn, 2002), the dioxin-binding affinity of the harbor seal AHR was at least as high as that of the AHR from a dioxin-sensitive strain of mice, suggesting that this seal species may be sensitive to PHAH effects. This implies that Baikal seal may also be sensitive to dioxin effects.

Amino Acid Sequence↗

Genome sequence based, comparative analysis of the fluorescent amplified fragment length polymorphisms (FAFLP) of tubercle bacilli from seals provides molecular evidence for a new species within the Mycobacterium tuberculosis complex.

Tuberculosis in seals is caused by a member of the Mycobacterium tuberculosis complex referred to as the 'seal bacillus'. Fluorescent amplified-fragment length polymorphism (FAFLP) analysis was applied to isolates from four Australian and six Argentinean seals and compared with FAFLP pattern for standard strains belonging to the M. tuberculosis complex. The FAFLP profiles derived from EcoRI/MseI restricted fragments of blind coded DNA samples differentiated the seal bacillus from other members of the M. tuberculosis complex. According to the phylogenetic analysis performed using FAFLP data, seal bacilli appear to have diverged significantly from other members of the M. tuberculosis complex. We describe the suitability of a panel of 19 highly polymorphic markers for rapid identification and comparative genomic analyses of the seal bacillus strains. It is likely that these bacilli got separated from the M. tuberculosis lineage as a result of different insertion deletion events occurring on a genome wide scale. Our analysis reveals that the seal bacillus and M. bovis are genetically related and therefore, might have originated from a common ancestor. Our data additionally support the hypothesis that seal bacillus occupies a unique taxonomic position within the M. tuberculosis complex.

Animals↗

Comparison of toxaphene congeners levels in five seal species from eastern Canada: what is the importance of biological factors?

Environmentally relevant chlorobornanes (CHBs) were measured in blubber samples of harbor (Phoca vitulina), gray (Halichoerus grypus), harp (Phoca groenlandica), and hooded seals (Cystophora cristata) sampled in different part of the St. Lawrence marine ecosystem (SLME) and ringed seals (Phoca hispida) sampled in the eastern Canadian Arctic waters. The purpose of this study was to compare the levels of six CHBs (Parlar-26, -40/-41, -44, -50, and -62) among the five seal species. Seal species could be separated into three groups based on their respective sigmaCHB mean concentrations (+/-standard error): gray (49+/-3.9 ng/g lipid weight) and harbor (80+/-20 ng/g lipid weight) seals were more contaminated than ringed seals (18+/-7.6 ng/g lipid weight) but less contaminated than harp (370+/-87 ng/g lipid weight) and hooded (680+/-310 ng/g lipid weight) seals. These differences are not expected to be related to different sources of toxaphene contamination, since both the SLME and the eastern Canadian Arctic environments are thought to be mainly contaminated via atmospheric transportfrom the southeastern part of the United States. Thus, biological factors such as sex, age, nutritive condition, metabolism capacity, and diet of the animals collected were considered. Results reported in this study indicated that the diet is likely the main factor accounting for interspecies variations in toxaphene contamination in seals from eastern Canada.

Adipose Tissue↗

Microsatellite variation in ringed seals (Phoca hispida): genetic structure and history of the Baltic Sea population.

Genetic variability and population structure of Baltic ringed seals and an Arctic reference population were assessed using eight microsatellite loci. Ringed seals colonized the Baltic Sea basin soon after deglaciation 11 500 years ago and are supposed to have remained largely isolated from the main Arctic stock since then, approximately 1000 generations. In the 1900s the Baltic population declined rapidly, and is now confined to three distinct breeding areas, with N < 6000 seals altogether. Microsatellite heterozygosity in ringed seals was higher than that in the closely related, boreal harbour seal and grey seal, for which the markers were initially developed. This is plausibly attributed to an overall greater population (species) size of ringed seals during the Quaternary. Allele frequency differentiation between the Baltic and Arctic ringed seals, conventionally treated as different subspecies, was weak. Assuming complete isolation, the divergence (FST=0.023) would imply a notably high postglacial effective population size, approximately 20 000 for the Baltic population. The isolation assumption however, seems unrealistic in the light of the data: a coalescent-based simulation approach to the likelihood of alternative demographic histories clearly favoured a scenario with recurrent gene flow to the Baltic, over one of complete isolation (drift only). Within the Baltic Sea, no differentiation was found between the Gulf of Finland and the Gulf of Bothnia breeding areas; the recent population decline and split have not yet affected the inbreeding levels of the disjunct breeding stocks.

Alleles↗

Heart rate and oxygen consumption of northern elephant seals during diving in the laboratory.

Many techniques have been employed to measure metabolic and cardiovascular changes in diving marine mammals. Each of these methods has its advantages, but the methods also have drawbacks when applied to phocid seals. The aim of this study was to investigate heart rate and metabolic responses to diving in juvenile northern elephant seals that are not associated with forced changes in exercise state, and, secondarily, to investigate whether heart rate could be used as an indicator of metabolic rate in this species. Six seals were allowed to dive freely in a metabolic chamber while simultaneous measurements of heart rate and oxygen consumption were made. Within each dive cycle (dive and surface interval), the seals spent an average of 74% of the time submerged. Mean dive duration was 6.43+/-0.6 (SD) min. Mean oxygen consumption during diving was 3.32+/-0.4 mL O2 min-1 kg-1, a decrease of approximately 26% from baseline values. An inverse relationship was observed between oxygen consumption and the percentage of time spent submerged in each dive cycle. The total amount of oxygen consumed during the surface interval increased with increasing dive duration, while the duration of the surface interval itself did not change, indicating that seals alter the rate of O2 uptake rather than the time spent at the surface. Mean heart rate during diving was 34.5+/-6.2 beats min-1, 36% lower than resting values. Mean diving heart rate was independent of dive duration, percent time submerged, and oxygen consumption. Mean surface interval heart rate was 66.6+/-11.1 beats min-1 and was not correlated with oxygen consumption. Average heart rate over the entire dive cycle increased with increasing oxygen consumption in all of the seals, but there was only a significant relationship in two seals, which casts some doubt on the usefulness of heart rate as an indicator of metabolic rate in this species. While providing important information on the changes in heart rate and oxygen consumption during diving in northern elephant seals, a complete understanding of the diving metabolic rate of these animals will require a combination of approaches that can be used in concert with data on freely living animals.

Animals↗

Identification and real-time PCR quantification of Phocine distemper virus from two colonies of Scottish grey seals in 2002.

The North Sea European harbour seal (Phoca vitulina) population has endured two phocine distemper virus (PDV) epidemics in 1988 and 2002. The grey seal (Halichoerus grypus) is a sympatric seal species that shows little or no mortality from PDV. Two Scottish grey seal breeding colonies were sampled for evidence of PDV infection approximately 2 months after the peak of the 2002 epidemic. In both colonies, a proportion of mothers (13/109) and pups (6/84) tested positive for PDV in their leukocytes. All infected animals were asymptomatic and completed the breeding season successfully. These results illustrate that grey seals come into contact with infectious seals and can become infected themselves without experiencing acute effects. In some seals the virus is able to replicate from the primary site of infection. This study provides evidence that grey seals may have an active role in the spread of PDV during an epidemic.

Animals↗

Onboard acoustic recording from diving northern elephant seals.

This study was the first phase in a long-term investigation of the importance of low-frequency sound in the aquatic life of northern elephant seals, Mirounga angustirostris. By attaching acoustic recording packages to the backs of six translocated juveniles, the aim was to determine the predominant frequencies and sound levels impinging on them, and whether they actively vocalize underwater on their return to their rookery at Ano Nuevo, California, from deep water in Monterey Bay. All packages contained a Sony digital audio tape recorder encased in an aluminum housing with an external hydrophone. Flow noise was minimized by potting the hydrophone in resin to the housing and orienting it posteriorly. The diving pattern of four seals was recorded with a separate time-depth recorder or a time-depth-velocity recorder. Good acoustic records were obtained from three seals. Flow noise was positively correlated with swim speed, but not so high as to mask most low-frequency sounds in the environment. Dominant frequencies of noise impinging on the seals were in the range 20-200 Hz. Transient signals recorded from the seals included snapping shrimp, cetacean vocalizations. boat noise, small explosive charges, and seal swim strokes, but no seal vocalizations were detected. During quiet intervals at the surface between dives, the acoustic record was dominated by respiration and signals that appeared to be heartbeats. This study demonstrates the feasibility of recording sounds from instruments attached to free-ranging seals, and in doing so, studying their behavioral and physiological response to fluctuations in ambient sounds.

Acoustics↗

Coronary blood flow and myocardial segment dimensions during simulated dives in seals.

Three harbor seals Phoca vitulina richardsi and five spotted seals Phoca vitulina largha were used in studies of acute episodes of local myocardial ischemia in open-chest, anesthetized animals and of coronary blood flow and regional function as indicated by left ventricular segment dimensions during experimentally simulated dives of conscious, instrumented animals. We observed that seal myocardium, in which there are few coronary anastomoses, responded to brief local occlusion with prompt local dysfunction and systolic bulging; coronary flow in the nondiving seal oscillated irregularly and declined with spontaneous apnea and related falling heart rate; flow continued to oscillate but was much reduced during dives, frequently ceasing entirely for periods as long as 45 s; ventricular segment dimension shortening was reduced intermittently during dives; and elevated heart rate induced during dives by cardiac pacing or by administration of atropine diminished or eliminated the reductions in coronary blood flow. Responses of seal heart reflect the reduction in cardiac metabolic demand during diving and the seal's myocardial adaptation for enhanced anaerobic glycolysis. The seal heart can maintain mechanical function during dives with minimal coronary perfusion, despite the progressive and ultimately profound hypoxia, hypercapnia, and acidosis. Reduced cardiac metabolism, copious glycolytic reserves, and metabolite washout by intermittent brief bursts of coronary blood flow are apparently sufficient to support continued cardiac function, even though the seal heart has little tolerance for acute localized ischemia.

Animals↗

Volume capacity and contraction control of the seal spleen.

Volume changes in the spleens of hooded seals (Cystophora cristata) and harp seals (Phoca groenlandica) were measured plethysmographically in vitro in response to epinephrine, norepinephrine, isoprenaline, phentolamine, and acetylcholine. Dilated spleens contracted forcefully within 1-3 min of alpha-adrenoceptor activation with 1.0-5.0 micrograms epinephrine/kg body mass, whereas stimulation of beta-adrenoceptors and cholinergic receptors had little effect. The mass of dilated hooded seal spleens corresponded to 2-4% (n = 7) of body mass, with volume (V; ml) relating to body mass (M; kg) as follows: V = 12.0M + 910 (r2 = 0.96, n = 4). Thus the spleen of a 250-kg hooded seal maximally expels 3.9 liters, or 13%, of its estimated total blood volume. Average hematocrit in splenic venous outflow from dilated spleens was 90 +/- 3% (n = 3) in hooded seals and 85% (n = 2) in harp seals. From these data we have estimated that the aerobic diving limit of a 250-kg hooded seal increases only 105 s, at the most, if complete emptying of the spleen occurs during diving, while the corresponding estimate for a 112-kg harp seal is 80 s.

Animals↗

Factors affecting stroking patterns and body angle in diving Weddell seals under natural conditions.

Aquatic animals use a variety of strategies to reduce the energetic cost of locomotion. Efficient locomotion is particularly important for breath-holding divers because high levels of exercise may quickly deplete oxygen reserves, leading to the termination of a dive. We investigated the swimming behavior of eight adult Weddell seals, which are proficient divers, in McMurdo Sound, Antarctica. A newly developed data logger was attached to free-ranging females at their own breeding sites to record swimming speed, depth, two-dimensional accelerations (stroke frequency and body angle) and temperature. All seals conducted multiple deep dives (the mean dive depth range for each animal was 223.3+/-66.5-297.9+/-164.7 m). Prolonged gliding while descending was observed with thinner females (N=5 seals). But the fatter females (N=3 seals) exhibited only swim-and-glide swimming, characterized by intermittent stroking and fluctuating swim speed, throughout their descent and ascent. The body angles of four of the seals were restricted to less than 30 degrees by the location of breathing holes in the ice and the slope of local bathymetric features. Of these four, the three fatter seals adopted the stroke-and-glide method while the other thinner seal descended with prolonged periods of gliding. Prolonged gliding seems to be a more efficient method for locomotion because the surface time between dives of gliding seals was significantly less than that of stroking animals, despite their same stroke frequencies.

Acceleration↗

Clinical and pathological characterization of northern elephant seal skin disease.

From 1984 through 1992, staff at The Marine Mammal Center (TMMC, Sausalito, California, USA) examined 207 northern elephant seals (Mirounga angustirostris) with a condition of unknown etiology called northern elephant seal skin disease (NESSD). The skin lesions were characterized by patchy to extensive alopecia and hyperpigmentation, punctate or coalescing epidermal ulceration, and occasionally, massive skin necrosis. Microscopic lesions included ulcerative dermatitis with hyperkeratosis, squamous metaplasia and atrophy of sebaceous glands. All diseased seals were less than 2 years of age and suffered from emaciation, depression, and dehydration. Mortality from septicemia increased significantly with severity of skin ulceration. Compared to 14 apparently unaffected seals, diseased seals had depressed levels of circulating thyroxine, triiodothyronine, retinol, serum iron, albumin, calcium, and cholesterol. Levels of alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase, gamma glutamyl transpeptidase, blood urea nitrogen, and uric acid were elevated. Morphometrically, diseased animals were approximately 15% smaller than normal seals of the same sage. Serum and blubber concentrations of 36 polychlorinated biphenyl congeners (sigma PCB) and dichloro-diphenyl-dichloroethylene (p,p'-DDE) were negatively correlated with body mass. Mean concentrations of sigma PCB and p,p'-DDE in serum in diseased seals were elevated as compared to apparently normal seals. Etiology of this syndrome remains unknown, but the possibility of PCB toxicosis cannot be ruled out.

Alopecia↗

Leptospirosis in rehabilitated Pacific harbor seals from California.

Renal disease was observed in two rehabilitated Pacific harbor seals (Phoca vitulina richardsii) from a facility in California (USA). The seals had leukocytosis and high serum phosphorus, blood urea nitrogen and creatinine concentrations. A retrospective study of leptospiral antibody serum titers indicated both seals had elevated titers to Leptospira interrogans serovar grippotyphosa. A third seal, which died about the time when the index cases occurred, also had elevated titers to L. interrogans serovar grippotyphosa. Post mortem histopathologic examination of all three seals showed tubular necrosis consistent with interstitial nephritis; spirochetes were seen within the kidney parenchyma of the third seal. Sea lions (Zalophus californianus) or elephant seals (Mirounga angustirostris) housed near the harbor seals were possible sources of exposure, but local wildlife also could have been responsible.

Animals↗

Leptospirosis in northern elephant seals (Mirounga angustirostris) stranded along the California coast.

Leptospirosis was identified in six northern elephant seals (Mirounga angustirostris) that were stranded in 1995 along the coast of California (USA). Histologic lesions in all seals included tubulointerstitial nephritis with tubular degeneration and necrosis. Infection was confirmed through identification of spirochetes using an immunohistochemical stain for Leptospira sp. antigens. One affected seal had an elevated titer to Leptospira interrogans serovar pomona. Four of the six seals developed leptospirosis during rehabilitation, and two seals had evidence of exposure in the wild. Potential sources of infection during rehabilitation include other elephant seals, California sea lions (Zalophus californianus), Pacific harbor seals (Phoca vitulina richardsii), or free-ranging wildlife. These results indicate that northern elephant seals are susceptible to leptospirosis and can develop disease both in the natural environment and in a rehabilitation setting.

Animals↗