Characterization of Otostrongylus circumlitus from Pacific harbor and northern elephant seals

Otostrongylus circumlitus (Railliet, 1899) from Pacific harbor seals (Phoca vitulina richardsi) and northern elephant seals (Mirounga angustirostris) were examined using morphological and molecular methods to determine whether northern elephant seals along the central California coast are infected by the same species of Otostrongylus as are Pacific harbor seals in the same area. Fixed nematodes were examined and measured using light microscopy. The polymerase chain reaction (PCR) was used to amplify and sequence the second internal transcribed spacer (ITS-2) and D3 expansion (26S) regions of ribosomal DNA of O. circumlitus from Pacific harbor and northern elephant seals. The ITS-2 region was also amplified from Parafilaroides sp. from the Pacific harbor seal, northern elephant seal, and California sea lion (Zalophus californianus) and used for restriction fragment length polymorphism (RFLP) analysis. Morphologically, it was not possible to distinguish O. circumlitus from Pacific harbor and northern elephant seals, and over a consensus length of 443 base pairs (bp) for ITS-2 and 321 bp for D3 the sequences of O. circumlitus from both hosts were identical. With the PCR-RFLP assay, it was possible to distinguish O. circumlitus from Parafilaroides sp. The results suggest that O. circumlitus is the same species in Pacific harbor and northern elephant seals, and molecular methods make it possible to distinguish this nematode from related nematodes.     

Immunoglobulin responses of northern elephant and Pacific harbor seals naturally infected with Otostrongylus circumlitus

Immunoglobulin (Ig) binding patterns of Pacific harbor seals (PHS, Phoca vitulina richardsi) and northern elephant seals (NES, Mirounga angustirostris) to tissues of adult Otostrongylus circumlitus were examined by immunoblotting to investigate the role of age in the unusual response of juvenile NES to infection with O. circumlitus. Serum was taken from NES between March 1997 and March 2001 and from PHS between May 1996 and August 1999. The serum of seals infected with O. circumlitus contained antibodies that bound to all nematode tissues examined. Intensity of band staining on Western blots suggested that there were higher levels of antibody recognizing the excretory-secretory (ES) glands in the serum of NES that were 1 yr and older and in the majority of PHS compared with that in 2- to 9-mo-old NES. All juvenile NES infected with O. circumlitus and a proportion of the PHS and older NES infected with O. circumlitus contained Ig specific to a 28 kDa protein band that was dominant in the female reproductive tract of the nematode. The Ig binding patterns of NES and PHS to adult Parafilaroides sp., larval Pseudoterranova sp., and larval and adult Anisakis sp. differed sufficiently from that of O. circumlitus that immunoblotting for the 28 kDa protein could be useful for diagnosis of this parasite in juvenile NES. The banding patterns suggest that O. circumlitus nematodes die and disintegrate in PHS and NES and that NES of 1 yr and older and most PHS respond differently to the ES glands than 2- to 9-mo-old NES.     

Infection with a novel gammaherpesvirus in northern elephant seals (Mirounga angustirostris)

Twenty juvenile northern elephant seals (Mirounga angustirostris) that died between 1998 and 2004 had ulcers on the tongue, palatine mucosa, and/or tonsils. Histologic examination of the lesions revealed cytoplasmic swelling, nuclear pyknosis, and eosinophilic to amphophilic intranuclear inclusions bodies suggestive of herpesviral infection. Electron microscopic examination and polymerase chain reaction analysis confirmed the presence of a herpesvirus. Subsequent DNA sequencing identified this to be a new gammaherpesvirus that was similar to Porcine lymphotropic virus 2, Alcephaline herpesvirus 1 (malignant catarrhal fever virus from wildebeest), and Chlorocebus rhadinovirus 1 from African green monkeys. Identical herpesviral DNA was also detected in blood and mucosal swabs collected from five healthy elephant seal pups.     

Prevalence and pathology of nematode infections in the lungs of ringed seals (Phoca hispida) of the western arctic of Canada

Two genera of lung nematodes were found in a sample of 382 wild ringed seals (Phoca hispida). Otostrongylus circumlitus were found mainly in young of the year where they occupied the main stem bronchi causing extensive mucus production, mucosal hyperplasia and peribronchitis. Immature stages of the nematode were found in pulmonary vessels causing end-arteritis. There was only a slight reduction in respiratory parenchyma in infected seals and no correlation between infection and size or body condition. However, the sharp decline of O. circumlitus infection from 32% in young of the year to 0.5% after the first winter suggests that this parasite may play a role in the population dynamics of ringed seals. Filaroides (Parafilaroides) hispidus was found in alveoli of seals throughout all age classes causing no significant lesions. The infection rate increased from 44 to 71% in the 7 to 13-yr-old animals and declined to 38% in older seals. Concurrent infections occurred in 28% of young of the year seals.     

Persistent right aortic arch and cribiform plate aplasia in a northern elephant seal (Mirounga angustirostris)

A female weanling northern elephant seal (Mirounga angustirostris) presented to The Marine Mammal Center in Sausalito, California, USA, in poor body condition. An esophageal obstruction was diagnosed by contrast radiography and esophagoscopy, but despite extensive diagnostics and supportive care, the seal died 6 days later. On postmortem examination, the right aortic arch was persistent, forming a vascular ring anomaly with a patent ductus arteriosus that compressed the distal esophagus. Aplasia of the right cribiform plate and hypoplasia of the right olfactory nerve was also identified. A review of necropsy reports from January 1988 to December 2003 revealed 16 severe congenital anomalies in 454 juvenile northern elephant seals that stranded in northern California.     

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.     

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.     

Parasites and associated pathology observed in pinnipeds stranded along the Oregon coast

Forty-two seals and sea lions found dead along the Oregon Coast were examined for parasites and associated pathology. Nematode infections of the lung and/or gastrointestinal tract were the primary cause of death in 5 of 42 animals examined. New distribution records were established for Pricetrema zalophi and Zalophotrema hepaticum. New host records include Z. hepaticum and Diphyllobothrium cordatum in the Steller's sea lion (Eumetopias jubatus); Nanophyetus salmincola in the California sea lion (Zalophus californianus); P. zalophi in the harbor seal (Phoca vitulina); and P. zalophi, Trigonocotyle sp. and Otostrongylus circumlitus in the northern elephant seal (Mirounga angustirostris).     

Development of foraging behavior in juvenile northern elephant seals

Rapid development of foraging ability is critical for phocids. In northern elephant seals Mirounga angustirostris , juvenile survivorship is low compared with adults and foraging difficulties are potentially associated with increased mortality. At Año Nuevo, California, foraging behavior of nine juvenile females during their third foraging migration and five juvenile females on their fourth foraging migration were documented using a variety of commercially available and custom time depth recorders. Foraging success, diving ability, time at depth, bouts of behavior and body composition changes were compared between trips to sea. There were no significant differences in foraging success measured as mass gain between the third and fourth trips to sea. There were differences in how energy was deposited between lean and adipose tissue compartments. Diving ability developed between trips to sea, reflected in significant increases in depth, dive duration and bottom time. Development also occurred within trips to sea. Depth, dive duration and bottom time increased with time at sea. Aerobic capacity appears to increase between the third and fourth trip, with a significantly increased percentage of total time submerged and a significantly lower diving rate. All juveniles on the fourth trip and four out of nine juveniles on the third trip followed marked diel patterns, foraging deep during the day and shallow at night. Like adults, juveniles appeared to stay primarily aerobic with surface intervals independent of dive durations. These results confirm that female juvenile northern elephant seals undergo important developmental changes in foraging behavior between the third and fourth trip, but these changes do not significantly impact foraging success.     

Rate of passage of digesta through the alimentary tract of southern elephant seals (Mirounga leonina) (Carnivora: Phocidae)

The inert markers chromium-EDTA (liquid phase marker) and ytterbium nitrate (solid phase marker) were added to the food of three southern elephant seals in Taronga Zoo, Sydney, in a series of nine trials. The enclosures were checked at 15 to 30 minute intervals for up to 60 hours after dosing, and all faeces voided on land were collected (91 samples). Marker concentrations in faecal dry matter were determined and mean retention times calculated from the concentration-time curves.
The faeces were soft to semiliquid, with mean water content of 58% (range 24–80%). The marker concentration curves indicated a rapid rate of food transit through the gastro-intestinal tract in elephant seals compared with other carnivores. The mean time between dosing and first recovery of marker (Initial Recovery Time) was nine hours. This was significantly longer than the figure of 4.8 hours for northern elephant seals reported previously, and possible reasons for the differences are discussed. Mean Retention Time, a better index of rate of passage of ingesta, was 13 hours for the three southern elephant seals. This compares with times of 22 hours for the dog, 15 hours for the raccoon, and 13 hours for the cat, all carnivores with much shorter gastrointestinal tracts, both absolutely and relative to body size, than the southern elephant seal.
It is suggested that the very long small intestine may be an adaptation to foraging at depth, combined with long periods of submergence and the need to ingest large amounts of food when the animal is at sea continuously for weeks or months.     

Angiotensin II and aldosterone increase with fasting in breeding adult male northern elephant seals (Mirounga angustirostris)

The renin-angiotensin-aldosterone system (RAAS) appears to contribute significantly to osmoregulation of fasting northern elephant seal (Mirounga angustirostris) pups; however, RAAS has not been characterized in fasting adult seals. Therefore, this study examined the contribution of RAAS to water turnover rates in fasting adult male northern elephant seals. Blood samples were obtained twice during their breeding fast at an interval of 6.5 wk, and water efflux rate was estimated by isotopic dilution during the same period. Serum electrolytes (Na+, K+, Cl-) and osmolality were unaltered between the two sampling periods, indicating ionic and osmotic homeostasis during the fast. Despite the lack of an increase in vasopressin, serum angiotensin II and aldosterone were increased and were significantly and positively correlated. Changes in aldosterone concentration and water efflux rate were significantly and negatively correlated, suggesting that the greater the increase in aldosterone, the smaller the loss of water. Adult male seals maintain ionic and osmotic homeostasis similar to that of fasting weaned pups, and this homeostasis appears to be mediated, at least in part, by RAAS, which probably contributes to increased water retention as well. The hormonal mechanisms by which northern elephant seals maintain water and electrolyte balance during fasting conditions appear to be similar regardless of age.     

Plasma melatonin concentration in neonatal northern elephant seals,Mirounga angustirostris

The development of pineal function in northern elephant seals was examined in an attempt to understand the physiological basis for previously observed high daytime levels of melatonin in neonatal southern elephant seals. Pineal glands from four northern elephant seal pups, estimated age less than 1 week, weighed 3.0 ± 0.80 g, which was significantly less than that previously found in southern elephant seals (4.6 ± 0.35 g). Midday concentrations of plasma melatonin in pups averaged more than 3000 pmol/l in the first 5 days post-partum, but declined rapidly to less than 400pmol/l after day 9. Daytime melatonin levels in northern elephant seals tended to be lower than in southern elephant seals, although they were very high compared with other species. A circadian cycle of plasma melatonin concentration was observed in newborn northern elephant seals, with levels of 3000–5000 pmol/1 during the day, rising to more than 10,000 pmol/1 late in the dark phase. Soon after weaning at 4 weeks of age, daytime and night-time levels were in the range 60–100 pmol/1 and 100–400 pmol/1, respectively. When approximately 10 weeks old, most samples were in the range 100–400 pmol/1 with no discernible difference between day and night levels. The results do not support the hypothesis that the pineal gland is involved in thermogenesis in new-born southern elephant seals. Instead, the very active pineal gland may contribute to energy conservation, by lowering body temperature, particularly at night. As physical insulation is acquired by the deposition of blubber, the mechanism is not required and melatonin falls to adult levels.     

HOMING BEHAVIOR OF JUVENILE NORTHERN ELEPHANT SEALS

The aim of this study was to determine if juvenile northern elephant seals, Mirounga angustirostris , translocated from their rookery would return to it quickly and reliably. During the spring and fall of 1990 and 1991 we captured 75 seals at Año Nuevo State Reserve, CA, U. S. A. and translocated them to release sites up to 100 km away. Eighty-eight percent of the seals returned to the capture site within 4.7 ± 4.3 d. Homing rate increased with age, but even the youngest seals (8–10 mo) homed at a 73% rate. Homing rate did not vary significantly with sex, season, or year. Data from diving instruments suggested that the seals often followed direct routes home, arrived on the rookery significantly more often at night than during the day, and when released together, returned separately. Mean homing speed of 18 seals with complete diving records was 39 km/d (range 3–70 km/d). Instruments on seals had no detectable effect on homing rate or homing speed. The translocation paradigm provides a powerful tool for conducting intensive shortterm studies on free-ranging seals.     

Long-term monitoring of Antarctic pinnipeds in Admiralty Bay (South Shetlands, Antarctica)

Year-round monitoring of five Antarctic pinnipeds was conducted in Admiralty Bay from 1988 up to 2000. Two breeding species: southern elephant sealsMirounga leonina (Linnaeus, 1758) and Weddell sealsLeptonychotes weddellii (Lesson, 1826), were present throughout the year. Three other species: crabeater seals Lobodon carcinophagus (Hobron and Jacquinot, 1842), leopard sealsHydrurga leptonyx (Blainville, 1820), and Antarctic fur sealsArctocephalus gazella (Peters, 1875) visited the area only for short periods. During this study, the abundance of elephant seals was stable, whereas those of Weddell and crabeater seals declined. Leopard seals numbers fluctuated irregularly. We detected a possible immigration from South Georgia: of a stable magnitude for elephant seals, and of variable magnitude, depending on food accessibility, for Antarctic fur seals. We found a strong recurrence of the spatial distributions of elephant, Weddell, and Antarctic fur seals in the 13 oases on the shore of Admiralty Bay. Annual distribution patterns were characteristic for each species. The innermost beaches were used predominantly by the animals during their annual fasts: the breeding and the moulting seasons.     

Body mass loss of moulting female southern elephant seals,Mirounga leonina,at Macquarie Island

Thirteen female southern elephant seals moulting at Macquarie Island lost an average of 4.46±0.80 kg/day (10.01±1.20g/kg/day). There was no significant difference between this rate of body mass loss and that reported for moulting female southern elephant seals from South Georgia. Moulting female southern elephant seals however exhibited larger mass specific mass loss than either female northern elephant seals or male southern elephant seals, indicating a higher metabolic cost of moult in these animals.     

Lipid composition of blood platelets and erythrocytes of southern elephant seal (Mirounga leonina) and antarctic fur seal (Arctocephalus gazella)

Erythrocyte and blood platelet phospholipid compositions were studied in three elephant seals and two fur seals, two species of marine mammals living in the Subantarctic region feeding on preys rich in (n-3) polyunsaturated fatty acids. Results were compared with those reported for related species and humans. In erythrocytes, the phospholipid (PL) and cholesterol (CHOL) contents were lower in pinnipeds than in humans. Phosphatidylcholine (PC) levels were higher in elephant seals than in fur seals, with a reverse trend for phosphatidylethanolamine (PE) and phosphatidylserine (PS). Both species had lower SM/PC ratios and PE plasmalogen concentrations than human. Erythrocytes were richer in (n-3) fatty acids (FA) in pinnipeds than in humans. In platelets, the PL content was lower and the CHOL content higher in elephant seals than in humans or in other phocid seal species studied to date. The SM/PC ratio was much higher than in other seal species or in man. In both species, the proportion of PE plasmalogens was higher in platelets than in erythrocytes. PL were more saturated in elephant seals than in fur seals. These results suggest that the erythrocytes and platelets of wild marine mammals may prove useful models to study the influence of dietary lipids on the structure and hemostatic function of these cells.     

Molecular systematics of pinniped hookworms (Nematoda: Uncinaria): species delimitation,host associations and host-induced morphometric variation

Hookworms of the genus Uncinaria have been widely reported from juvenile pinnipeds, however investigations of their systematics has been limited, with only two species described, Uncinaria lucasi from northern fur seals (Callorhinus ursinus) and Uncinaria hamiltoni from South American sea lions (Otaria flavescens). Hookworms were sampled from these hosts and seven additional species including Steller sea lions (Eumetopias jubatus), California sea lions (Zalophus californianus), South American fur seals (Arctocephalus australis), Australian fur seals (Arctocephalus pusillus), New Zealand sea lions (Phocarctos hookeri), southern elephant seals (Mirounga leonina), and the Mediterranean monk seal (Monachus monachus). One hundred and thirteen individual hookworms, including an outgroup species, were sequenced for four genes representing two loci (nuclear ribosomal DNA and mitochondrial DNA). Phylogenetic analyses of these sequences recovered seven independent evolutionary lineages or species, including the described species and five undescribed species. The molecular evidence shows that U. lucasi parasitises both C. ursinus and E. jubatus, whereas U. hamiltoni parasitises O. flavescens and A. australis. The five undescribed hookworm species were each associated with single host species (Z. californianus, A. pusillus, P. hookeri, M. leonina and M. monachus). For parasites of otarids, patterns of Uncinaria host-sharing and phylogenetic relationships had a strong biogeographic component with separate clades of parasites from northern versus southern hemisphere hosts. Comparison of phylogenies for these hookworms and their hosts suggests that the association of U. lucasi with northern fur seals results from a host-switch from Steller sea lions. Morphometric data for U. lucasi shows marked host-associated size differences for both sexes, with U. lucasi individuals from E. jubatus significantly larger. This result suggests that adult growth of U. lucasi is reduced within the host species representing the more recent host–parasite association. Intraspecific host-induced size differences are inconsistent with the exclusive use of morphometrics to delimit and diagnose species of Uncinaria from pinnipeds.     

Physiology and behavior influence lactation efficiency in northern elephant seals (Mirounga angustirostris)

The efficiency with which mothers convert acquired energy into milk is a key determinant of the magnitude of parental investment in mammals; however, the mechanisms underlying lactation efficiency are poorly understood. Investigations on northern elephant seals have shown lactation efficiency, measured as the proportion of total energy expenditure that goes to the pup as milk, increases with age. In a cross-sectional study the physiological and behavioral determinants of lactation efficiency were investigated in eight young and seven prime (older) elephant seals by conducting behavioral observations and collecting milk, blood, and tissue on days 3 and 22 of lactation. Milk composition, circulating fatty acid and triglyceride concentrations, and mammary and blubber lipoprotein lipase activity were determined. Prime females had significantly greater percent milk fat and circulating fatty acids on day 3 than did young females, but these differences disappeared by day 22. The ability for prime females to produce higher-energy milk early in lactation may allow them to increase lactation efficiency by increasing the rate of energy transfer. In addition, prime females spent significantly more time resting. A combination of reduced activity and more rapid energy delivery likely explains the increase in lactation efficiency with age found in a previous study.     

Hematologic and serum biochemical profile of the northern elephant seal (Mirounga angustirostris): variation with age, sex, and season

The foraging success, and thus the survival and reproductive success, of deep-diving pinnipeds such as the northern elephant seal, Mirounga angustirostris, depends on the ability to withstand repetitive breath-hold dives. Health parameters can be incorporated as potential explanatory variables for differences observed in diving and migratory performance of individual seals. Furthermore, biomedical samples from apparently healthy individuals can provide valuable baseline data for evaluating effects of natural or anthropogenic threats to individuals and populations. We evaluated 42 blood parameters in 134 northern elephant seals during the breeding and molting seasons (1992-1999) to test for age, sex, and seasonal differences and to develop reference ranges. Adult males sampled during the breeding season differed from all other adult groups for a suite of parameters often associated with inflammation, infection, or other stressors: lower hematocrit, higher white blood cell count, higher band neutrophils, higher neutrophil count, lower albumin, and lower serum iron. Adult females during the breeding season differed from all other adult categories for two parameters (lower platelet counts, lower alanine aminotransferase activity). Molting males had higher blood urea nitrogen than all other classes; creatinine did not differ between breeding and molting adult males, but was higher in males than in females in both seasons. We found significant differences among age classes for 24 of 42 parameters measured, including higher levels of triglycerides, total protein, calcium, and iron in pups than we found in juveniles or adults. Unlike other mammals which undergo substantial decreases in energy expenditure during prolonged fasting (e.g., hibernation), northern elephant seals defend territories, give birth and suckle large offspring, mate, and molt during their bi-annual fasts. Nonetheless, many studies have described physiologic homeostasis during fasting in elephant seals. The genus Mirounga is superbly adapted to going without feeding for extended periods, and this is reflected in our hematologic and serum biochemical profiles.