Histological,histochemical, and ultrastructural investigations on the gastrointestinal system of antarctic seals: Weddell seal (Leptonychotes weddellii) and crabeater seal (Lobodon carcinophagus)

The morphology of the principal sections of the gastrointestinal system of two Antarctic seals with different dietary habits, namely, the Weddell seal (Leptonychotes weddellii) and the crabeater seal (Lobodon carcinophagus), has been investigated. Histologically examined by light microscopy, the tissue layers of the gastrointestinal tract of both seals are almost identical to those observed in most other mammals and no major differences in principle organization could be found between the two seal species. The ultrastructure of the gastric and intestinal epithelial cells has been examined and is also closely comparable to that of these cells in other mammals; however, Paneth cells have not been found in our material. In general, therefore, adaptations of the gastrointestinal tract to the aquatic environment or the diet are not obvious at the morphological levels of organization studied. Histochemical differences are found between the two closely related species; mucins of the surface epithelium in the stomach of Weddell seals are highly sulfated, while those in the crabeater seal are not. Mucous neck cells in Weddell seals contain acid mucosubstances, while those of crabeater seals contain neutral ones. Goblet cells in the small and large intestine in Weddell seals contain both neutral and acid mucosubstances. Both mucin types are detected in the crabeater seal; however, the mucins of the colon in the crabeater seal are more highly sulfated than those in the Weddell seal. The ratio of globet cells to enterocytes in the large intestine of crabeater seals is higher than that in Weddell seals. © 1995 Wiley-Liss, Inc.     

A gross and microscopic study of the respiratory anatomy of the Antarctic Weddell seal, Leptonychotes weddelli.

Four species of Phocidae, or true seals, inhabit the waters surrounding the Antarctic continent. These animals are thought to have different diving capabilities. The Weddell seal, Leptonychotes weddelli, is known to be capable of attaining depths up to 600 meters. The respiratiory system of the Weddell seal shows the usual adaptations to an aquatic environment characteristic of other marine. These include lungs that undergo compression acollapse at depths greater than 70 meters; hyaline cartilage in the tracheo-bronchial tree as far as the terminal bronchioles; and large amounts of smooth muscle surrounding the distal-most bronchioles. The collapsible lungs provide a mechanism by which air is forced from the alveoli adjacent to the pulmonary capillary beds thereby preventing the absorption of nitrogen gas into the bloodsteam. The presence of hyaline cartilage throughout most of the tracheo-bronchial tree increases the effective dead air space that accommodates most of the air forced from the collapsed lungs. The smooth muscle surrounding the respiratory bronchioles prevents their collapse while under the pressures of a deep dive. Collapse of the respiratory bronchioles not supported by cartilage would trap air in the lung alveoli during a dive. In addition, large- sac-like "diverticulae" are found in the submucosa throughout the tracheo-bronchial tree. These diverticulae, which open directly into the lumen of the tree, appear to be modified glands whose cells, in most cases, do not appear to be specialized for secretory function. They are most numerous in the more distal bronchi and terminal bronchioles where they are situated on both the luminal and adventitial sides of the hyaline cartilage supporting the walls of the air passages. Diverticulae are not found in the respiratory bronchioles or in the respiratory portion of the lungs.     

Trophic interactions of Antarctic seals as determined by stable isotope signatures

The diets and trophic interactions among Weddell, crabeater, Ross, and leopard seals in the eastern Ross Sea, Antarctica, were investigated by the use of stable isotope techniques during the 1999–2000 summer seasons. The ¹³C and ¹⁵N values in seal serum clearly distinguished the three Antarctic pack-ice seal species at different trophic positions (Weddell>Ross>crabeater). These patterns appeared to reflect a close linkage to their known foraging ecology and diving behaviors, and agreed well with their presumed dietary diversity. The more enriched ¹³C and ¹⁵N values in male Weddell seals than those in females suggested differences in foraging preferences between them. Significant differences in ¹⁵N were also found among different age groups of Weddell seals. A strong correlation between the C:N ratios and serum cholesterol was probably due to extremely high cholesterol levels in phocids. Comparisons of isotope data with harbor seals revealed distinct differences between Antarctic phocids and the northern seal species.     

Histological, histochemical, and fine structural observations on the spleen of seals

Spleens of three species of Antarctic seals with different diving habits (Weddell seal, crabeater seal, and fur seal) have been studied with histological, histochemical, and electron microscopic methods. The spleens can be classified as nonsinusoidal, with capsule and trabeculae rich in innervated smooth muscle cells. The trabecular system is particularly well developed in the deep- and long-diving Weddell seal. As in other mammals the pulp can be divided into white and red pulp. In the white pulp, periarteriolar lymphatic sheaths and secondary lymphatic nodules occur; both are surrounded by a marginal zone rich in macrophages and eosinophils. The nodules can be observed frequently, which is in accordance with abundance of plasma cells in the red pulp. Well-developed white pulp and numerous plasma cells and eosinophils obviously reflect a high load of nematodes, which have mainly been found in lung and stomach. Additionally, in the red pulp morphological evidence for the following functions has been found: destruction of erythrocytes, erythropoiesis, and thrombopoiesis. In respect to blood flow through the red pulp, we interpret our observations in the following way: terminal branches of arterioles open into the space between the fibroblastic reticulum cells; blood draining from here is collected into pulp veins, which are mainly found near the trabeculae. Thus, the seals have an open vascular compartment in their spleens, as also occurs in the cat. The red pulp is innervated by numerous nerve fibers that seem to include both cholinergic and adrenergic ones. The target cells of these fibers seem to be the fibroblastic reticulum cells, whose state of contraction may influence the direction of blood flow through the red pulp.     

Pleistocene population expansions of Antarctic seals

We sequenced a portion ( c . 475 bp) of the mitochondrial control region of three species of Antarctic phocid carnivores (Weddell seal, Leptonychotes weddellii , N  = 181; crabeater seal, Lobodon carcinophaga , N  = 143; and Ross seal, Ommatophoca rossii , N  = 41) that live seasonally or permanently in the fast ice and seasonal pack ice of the western Amundsen and Ross seas of western Antarctica. We resolved 251 haplotypes with a haplotype diversity of 0.98 to 0.99. Bayesian estimates of Θ from the program LAMARC ranged from 0.075 for Weddell seals to 0.576 for crabeater seals. We used the values of theta to estimate female effective population sizes ( N_EF ), which were 40 700 to 63 000 for Weddell seals, 44 400 to 97 800 for Ross seals, and 358 500 to 531 900 for crabeater seals. We used mismatch distributions to test for historical population size expansions. Weddell seals and crabeater seals had significant, unimodal mean pairwise difference distributions ( P  = 0.56 and 0.36, respectively), suggesting that their populations expanded suddenly around 731 000 years ago (Weddell seals) and around 1.6 million years ago (crabeater seals). Both of these expansions occurred during times of intensified glaciations and may have been fostered by expanding pack ice habitat.     

Dinucleotide microsatellite markers from the Antarctic seals and their use in other Pinnipeds

Twenty‐four microsatellite loci were isolated from three species of Antarctic seals (Subfamily Monachinae, Tribe Lobodontini). Eleven loci were cloned from Weddell seal, Leptonychotes weddellii, seven from leopard seal, Hydrurga leptonyx, and six from crabeater seal, Lobodon carcinophagus. Variability was assessed in Weddell seals collected in McMurdo Sound, leopard seals from Bird Island, South Georgia, and crabeater seals sampled in the eastern Ross Sea. All loci were variable in the three species used for cloning and 22 of these loci amplified variable products in the Ross seal, Ommatophoca rossii. Cross‐species amplification was largely successful, with an average of 19 loci amplifying products in other phocids.     

Serum chemistry and antibodies against pathogens in antarctic fur seals, Weddell seals, crabeater seals, and Ross seals

Information on health parameters, such as antibody prevalences and serum chemistry that can reveal exposure to pathogens, disease, and abnormal physiologic conditions, is scarce for Antarctic seal species. Serum samples from Antarctic fur seals (Arctocephalus gazella, n=88) from Bouvet?ya (2000-2001 and 2001-2002), and from Weddell seals (Leptonychotes weddellii, n=20), Ross seals (Ommatophoca rossii, n=20), and crabeater seals (Lobodon carcinophagus, n=9) from the pack-ice off Queen Maud Land, Antarctica (2001) were analyzed for enzyme activity, and concentrations of protein, metabolites, minerals, and cortisol. Adult Antarctic fur seal males had elevated levels of total protein (range 64-99 g/l) compared to adult females and pups (range 52-79 g/l). Antarctic fur seals had higher enzyme activities of creatine kinase, lactate dehydrogenase, and amylase, compared to Weddell, Ross, and crabeater seals. Antibodies against Brucella spp. were detected in Weddell seals (37%), Ross seals (5%), and crabeater seals (11%), but not in Antarctic fur seals. Antibodies against phocine herpesvirus 1 were detected in all species examined (Antarctic fur seals, 58%; Weddell seals, 100%; Ross seals, 15%; and crabeater seals, 44%). No antibodies against Trichinella spp., Toxoplasma, or phocine distemper virus (PDV) were detected (Antarctic fur seals were not tested for PDV antibodies). Antarctic seals are challenged by reduced sea ice and increasing temperatures due to climate change, and increased anthropogenic activity can introduce new pathogens to these vulnerable ecosystems and represent a threat for these animals. Our data provide a baseline for future monitoring of health parameters of these Antarctic seal species, for tracking the impact of environmental, climatic, and anthropogenic changes in Antarctica over time.     

Blood rheology of Weddell seals and bowhead whales

Red blood cell (RBC) aggregation and blood viscosity are important determinants of in vivo blood flow dynamics and, in marine mammals, these parameters may impact diving physiology by altering blood oxygen delivery during the diving response. Weddell seals are superb divers and exhibit age-related patterns in blood oxygen chemistry and diving ability. By contrast, bowhead whales are not long duration divers, and little is known of their blood properties relative to diving. The present study was designed to compare rheological characteristics of blood from Weddell seal pups, Weddell seal adults, and from adult bowhead whales: blood viscosity and RBC aggregation in plasma and in polymer solutions (i.e., RBC "aggregability") were measured. Salient findings included: (1) significant 4- to 8-fold greater aggregation in blood from adult seals compared with pups and human subjects; (2) 2-to 8-fold greater aggregation in bowhead whale blood compared with human blood; (3) compared to human red cells, enhanced RBC aggregability of RBC from adult seals and whales as determined by their greater aggregation in polymer solutions; (4) increasing RBC aggregation and aggregability of seal pup blood over a seven day period following birth; (5) significantly greater blood viscosity for adult seals compared with pups at both native and standardized hematocrits. These results indicate that, for both species, hemorheological parameters differ markedly from those of humans, and suggest progressive changes with seal age; the physiological implications of these differences have yet to be fully defined.     

Architecture and functional aspects of the distal airways of Antarctic seals with different habits,the Weddell Seal (Leptonychotes weddellii) and the Crabeater Seal (Lobodon carcinophagus)

Summary The lung of the deep diving Weddell Seal is characterized by an unusually well developed periacinar dense collagenous connective tissue, and a thick coat of smooth musculature particularly in the distal bronchioli. Both, collagen and smooth musculature appear to be functionally interrelated, the first serving presumably as site of origin or attachment for the latter. The orientation of the bronchiolar smooth muscle cells is complex: there exists a basic pattern of two crisscrossing helical bundles that wind in opposite direction. In addition, longitudinal bundles are frequent both at the inside and the outside of the muscular coat. Furthermore, more or less complete ringshaped bundles occur as well as groups of muscle fibres running radially into the collagenous tissue of the surroundings of a bronchiolus. This complex architecture presumably allows active adjustment to various physiological needs of the Weddell Seal including as extremes both closing and widening of the bronchiolar lumen. Isometric contractions of the smooth musculature may stiffen the wall of the distal airways while diving. In the Crabeater Seal which dives for shorter durations and by far less deeply than the Weddell Seal, both periacinar collagen and bronchiolar smooth musculature are of similar arrangement, however, occur in considerably reduced amounts. A rich supply of autonomie nerve fibres with abundant varicosities controls the smooth muscle cells, which are interconnected by gap junctions and receive their innervation par distance (visceral type of smooth musculature). The majority of varicosities contains small clear vesicles, as is typical for cholinergic nerves, suggesting a strong parasympathetic influence. Other varicosities are presumably of peptidergic type. Mast cells and epithelial endocrine cells may exert additional influence on the musculature.     

Antibodies against european phocine herpesvirus isolates detected in sera of Antarctic seals

Summary Neutralizing antibodies against European phocine herpesvirus were detected in sera of to two Antarctic seal species, Weddell seals (Leptonychotes weddellii) and crabeater seals (Lobodon carcinophagus), collected in the eastern Weddell Sea. A large number of positive sera crossneutralized canine herpesvirus, but only few sera also contained antibodies to feline herpesvirus. The Weddell seals suffered from a respiratory disease when the sera were collected (January–February, 1990). The significance and possible origin of herpesvirus infections in Antarctic seals documented for the first time in this communication is discussed. All sera were negative for antibodies against phocine and canine distemper viruses.     

Identification of hairs found in leopard seal (<Emphasis Type="Italic">Hydrurga leptonyx</Emphasis>) scats

The leopard seal is a top-order predator in the Southern Ocean ecosystem and preys on a wide variety of vertebrate species including seals and penguins. We assessed the use of hairs found in leopard seal scats to identify the species of pinniped consumed. A reference collection of hairs was obtained from four potential leopard seal prey species including crabeater, Weddell, Ross, and Southern elephant seals. Discrimination techniques applied to terrestrial mammals did not allow for identification of the seal hairs. Instead, a 2-dimensional (2-D) and 6-dimensional (6-D) analysis technique utilising Mahalanobis distances (D ²) was used. The smallest Mahalanobis distance together with the largest value of p(F) positively identified hairs from each species. The 6-D analysis was more accurate and applied to hairs found in the leopard seal scats. The majority of prey species were identified as crabeater seals, which are a known prey item of the leopard seal.     

Genetically effective population sizes of Antarctic seals estimated from nuclear genes

We analyzed eight nuclear microsatellite loci in three species of Antarctic seals; Weddell seal (Leptonychotes weddellii; mean N = 163), crabeater seal (Lobodon carcinophaga; 138) and Ross seal (Ommatophoca rossii; 35). We estimated genetic diversity (Θ) and effective population size (N _E) for each species. Autosomal microsatellite based N _E estimates were 151,200 for Weddell seals, 880,200 for crabeater seals, and 254,500 for Ross seals. We screened one X-linked microsatellite (Lw18), which yielded similar N _E estimates to the autosomal loci for all species except the Ross seals, where it was considerably larger (~103 times). Microsatellite N _E estimates were comparable with previously published N _E estimates from mitochondrial DNA, but both are substantially lower than direct estimates of population size in all species except the Ross seals. The ratio of maternally versus biparentally derived estimates of N _E for Ross seals was not consistent with the hypothesis that they are a polygynous species. We found no sign of a recent, sustained genetic bottleneck in any of the species.     

Diving and haulout behavior of crabeater seals in the Weddell Sea,Antarctica, during March 1986

Summary Time-depth recorders were used to study the diving and haulout behavior of six crabeater seals in the marginal. ice edge zone of the Weddell Sea during March 1986. Haulout patterns revealed the seals' clear preference for diving during darkness and hauling out onto sea ice during daylight. Seals did not necessarily haul out every day; individual seals hauled out on 80–100% of days during the study period. Four general dive types were identified: 1) traveling dives, 2) foraging dives, 3) crepuscular foraging dives, and 4) exploratory dives. Nearly continual diving occurred for extended periods (about 16 h) nightly, with one individual diving up to 44 h without interruption. Foraging dives occurring during crepuscular periods were deeper than those made during the darkest hours. The authors suggest that the distinct diel pattern of dive timing and depth may be related to possible predator avoidance behavior by the seals' principal prey, Antarctic Krill.     

Brain size in neonatal and adult Weddell seals: Costs and consequences of having a large brain

Little is known about the ontogeny of brain size in pinnipeds despite potential functional implications of brain substrate (glucose, oxygen) requirements for diving, fasting, growth, and lactation strategies. We measured brain mass (brM) and cranial capacity (CC) in newborn and adult Weddell seals. Neonatal Weddell seals had brM that represented ~70% of adult brM. Weddell seals have the largest neonatal brain, proportional to adult brain, reported for any mammal to date, which is remarkable considering the relatively small size of Weddell seal pups at birth (6%–7% of maternal body mass) compared to neonates of other highly precocial mammals. Provision of sufficient glucose to maintain the large, well‐developed brain of the neonatal Weddell seal has a nontrivial metabolic cost to both pup and mother. We therefore hypothesize that this phenomenon must have functional significance, such as allowing pups to acquire complex under‐ice navigation skills during the period of maternal attendance.     

Gut length, food transit time and diving habit in phocid seals

The southern elephant seal (Mirounga leonina) has the ability to dive for 2 h and reach depths of 1200 m. This creature is also exceptional in having a small intestine that is 25 times body length. Krockenberger and Bryden advanced the hypothesis that the long small intestine has developed to compensate for the extended periods with reduced or even abolished intestinal blood perfusion during diving. To test this hypothesis we have measured small-intestinal lengths in crabeater (Lobodon carcinophagus), Weddell (Leptonychotes weddellii), Ross (Ommatophoca rossi), leopard (Hydrurga leptonyx), harp (Phoca groenlandica), ringed (Phoca hispida) and hooded (Cystophora cristata) seals and related them to available data on their maximal dive duration. We found no significant correlation (P > 0.05) between intestinal length relative to body length and diving ability, but we found that small-intestinal internal area was significantly (P < 0.05) related to body length. A crude scanning electron microscopical examination of the small intestines of Weddell, crabeater, hooded and harp seals failed to reveal any gross anatomical differences between small-intestinal surfaces. This suggests that gut dimension in this variety of phocid species with widely differing diving ability is not related to diving habit, but is instead related to body size. The transit time of digesta was determined in two 1-year-old harp seals by use of radiopaque polyethylene rings of 4-mm diameter followed by X-ray examination, as markers for the solid phase passage, and chromium ethylene-diaminetetra acetic acid (Cr-EDTA) as a marker for the liquid phase. The transit time for the Cr-EDTA marker was 6.9 h ± 0.5 SE (range 4.5–8 h, n= 7), while 80% of the polyethylene markers appeared in the colon after 17.6 h ± 1.0 SE (range 14–21.5 h, n= 6) and were sometimes retained in the colon for several hours before defecation. These transit times did not change significantly (P > 0.05) in response to repetitive diving over a period of 8 h. This indicates that the often-used Cr-EDTA is not a good measure for digesta passage time when used alone in seals, and that the hypothesis of Krockenberger and Bryden is most likely wrong. Received: 17 December 1997 / Accepted: 4 May 1998     

Observations on food remains in faeces of elephant,leopard and crabeater seals

Summary Faecal material of leopard, crabeater and elephant seals was collected from the vicinity of Davis station, Antarctica. Very few identifiable remains were found in elephant seal droppings. Fish remains, mainly of Pleuragramma antarcticum, were found in both leopard and crabeater seal droppings. The mysid Antarctomysis maxima was also found in crabeater seal droppings and amphipods and decapod crustaceans in leopard seal droppings.     

Diving physiology and winter foraging behavior of a juvenile leopard seal (Hydrurga leptonyx)

Diving physiology and at-sea behavior of a juvenile leopard seal (Hydrurga leptonyx) were opportunistically measured in the Antarctic Peninsula during winter 2002. Total body oxygen stores were estimated from measures of hematocrit, hemoglobin, myoglobin, and total blood volume and were used to calculate an aerobic dive limit (ADL). Movement patterns and diving behavior were measured by equipping the seal with a Satellite Relay Data Logger that transmitted data from 8–31 August 2002. The seal remained in a focal area, in contrast to crabeater seals tracked simultaneously. The seal displayed short, shallow dives (mean 2.0±1.4 min, 44±48 m) and spent 99.9% of its time within the estimated ADL of 7.4 min. The shallow diving behavior contradicts previous diet research suggesting Antarctic krill (Euphausia superba) is the primary prey of leopard seals during the winter months as krill were found at deeper depths during this period. These measurements of diving and movement of a leopard seal provide valuable preliminary data necessary to develop future research on the at-sea behavior of an apex predator in the Antarctic ecosystem.     

Contrasting behavior between two populations of an ice‐obligate predator in East Antarctica

The Austral autumn–winter is a critical period for capital breeders such as Weddell seals that must optimize resource acquisition and storage to provision breeding in the subsequent spring. However, how Weddell seals find food in the winter months remains poorly documented. We equipped adult Weddell seals after their annual molt with satellite‐relayed data loggers at two sites in East Antarctica: Dumont D'Urville (n = 12, DDU) and Davis (n = 20). We used binomial generalized mixed‐effect models to investigate Weddell seals’ behavioral response (i.e., “hunting” vs. “transit”) to physical aspects of their environment (e.g., ice concentration). Weddell seal foraging was concentrated to within 5 km of a breathing hole, and they appear to move between holes as local food is depleted. There were regional differences in behavior so that seals at Davis traveled greater distances (three times more) and spent less time in hunting mode (half the time) than seals at DDU. Despite these differences, hunting dives at both locations were pelagic, concentrated in areas of high ice concentration, and over areas of complex bathymetry. There was also a seasonal change in diving behavior from transiting early in the season to more hunting during winter. Our observations suggest that Weddell seal foraging behavior is plastic and that they respond behaviorally to changes in their environment to maximize food acquisition and storage. Such plasticity is a hallmark of animals that live in very dynamic environments such as the high Antarctic where resources are unpredictable.     

Some anatomical aspects of the cardiovascular system of antarctic seals and their possible functional significance in diving

The hearts and ascending aortae of 11 Weddell seals, Leptonychotes weddelli, three adult Crabeater seals, Lobodon carcinophagus, two adult Ross seals, Ommatophoca rossi, and one adult Leopard seal, Hydrurga leptonyx, were examined for comparison with terrestrial forms. The Weddell seal specimens were from animals ranging in age from midterm in fetal development to mature adults. All specimens were collected in 1971, 1972, and 1973, from McMurdo Sound and the Ross Sea, Antarctica. The phocid hearts were characteristically broader and flatter than those of other carnivore families and they tended toward bifid apices. The heart form indices (height/circumference) averaged 31.5 compared to 39.0 for felids. The right ventricular chambers of the Antarctic seals were found to average longer in Weddells and narrower in all, than those reported for four other carnivore families. An elastic enlargement was present in the ascending aortae of all seals. The largest diameter of the aortic bulb averaged 25.5 mm more than the base of the aorta in the adult Weddell seals which represented an increase of 72.5% over the base. It is suggested that the general heart form, and especially the ascending aortae, are anatomical adaptations to diving. The compressed heart makes possible unimpaired function when the chest is compressed during deep dives. The aortic bulb maintains mean arterial blood pressure and perfusion of the brain and cardiac tissue during diving bradycardia.     

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.