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PubMed1993

Epidermal lipid in several cetacean species: ultrastructural observations.

The ultrastructure of the skin of four cetacean species, bottlenose dolphin (Tursiops truncatus) long-finned pilot whale (Globicephala melaena), humpback whale (Megaptera novaeangliae), and fin whale (Balaenoptera physalus) was investigated with particular reference to epidermal lipid. It has already been established that massive lipid reservoirs exist in whales, that the biochemical structures of cetacean lipids are unique, and that unusual intracellular lipid droplets appear in the epidermis. We report here some novel findings on scanning electron microscopic morphology of epidermal lipid, and on its ultrastructural morphology in general and specialized integumentary sites, including species not previously investigated. The intracellular epidermal lipid droplets were more extensive than lamellar body-derived intercellular lipid which is within the interstices of stratum externum cells. The intracellular droplets were spherical, highly variable in size ranging from 0.24 micron to 3.0 microns in diameter, appeared singly or were aggregated in cytoplasmic cavitations, and often were closely associated with epidermal cell nuclei. Evidence for exocytosis of the intracellular droplets was not observed. Significant numbers of intracellular lipid droplets are not observed in the epidermis of terrestrial mammals, so their presence is one of several aquatic specializations of the cetacean integument. Its full significance remains obscure, but it is more probably associated with epidermal cell metabolism than with secretion of lipid.

Animals↗
PubMed1984

Conservation of highly repetitive DNA in cetaceans.

It is controversial whether odontocetes (toothed whales) and mysticetes (whalebone whales) have a common ancestry. Cetacean karyological uniformity, which is unique among mammalian orders, suggests a monophyletic origin; however, several anatomical authorities have maintained that odontocetes and mysticetes are diphyletic. We investigated the issue using Southern blot hybridization. Two labelled restriction fragment probes from the DNA of the sei whale (a mysticete) were hybridized to restricted DNA of cetacean species representing all extant families except the Eschrichtiidae, the gray whales. The probes hybridized to specific restriction fragments in all odontocete and mysticete materials. Hybridization showed preservation of hybridization homologies and a striking conservation of the length of highly repeated DNA sequences. The results are compatible with a common ancestry of odontocetes and mysticetes.

Animals↗
PubMed1987

Autoxidation of oxymyoglobin with the distal (E7) glutamine.

We reported previously that the distal(E7) histidine is replaced by glutamine in myoglobin from the shark, Galeorhinus japonicus. The amino-acid sequence of myoglobin from another shark, Heterodontus japonicus, has been determined. The myoglobin is composed of 148 residues, is acetylated at the N-terminus, and contains the distal(E7) histidine at position 59. Although the sequence homologies between G. japonicus, H. japonicus, and sperm-whale myoglobins were about 40-55%, their hydropathy profiles were very similar, indicating that they have a similar geometry in their globin folding. The autoxidation rates of the two shark oxymyoglobins were examined in 0.1 M buffer at 25 degrees C over pH range 4.5-11.5. The pH dependence for the autoxidation of H. japonicus myoglobin was very similar to that of sperm-whale myoglobin, although the rate was about 10-times higher over the pH range examined. In both myoglobins, autoxidation was largely accelerated by H+. On the other hand, the pH dependence of G. japonicus myoglobin, which has the distal glutamine in the place of histidine, was quite different from those of sperm-whale and H. japonicus myoglobins. One of the most remarkable features is the fact that the autoxidation rate is not enhanced with an increase in the concentration of H+ in the acidic range of pH, where the autoxidation of sperm-whale and H. japonicus myoglobins is most accelerated. This finding suggests that the distal(E7) histidine participates in the autoxidation reaction as a catalytic residue facilitating the movement of a catalytic proton.

Amino Acid Sequence↗
PubMed2005

Radiocaesium (137Cs) in marine mammals from Svalbard, the Barents Sea and the North Greenland Sea.

Specific activities of the anthropogenic radionuclide, 137Cs, were determined in marine mammals from Svalbard and the Barents and North Greenland Seas. Muscle samples were collected from 12 polar bears, 15 ringed seals, 10 hooded seals, 7 bearded seals, 14 harp seals, one walrus, one white whale and one blue whale in the period 2000-2003. The mean concentrations (+/-SD) of 137Cs were: 0.72+/-0.62 Bq/kg wet weight (w.w.) for polar bears; 0.49+/-0.07 Bq/kg w.w. for ringed seals; 0.25+/-0.10 Bq/kg w.w. for hooded seals; 0.22+/-0.11 Bq/kg w.w. for bearded seals; 0.36+/-0.13 Bq/kg w.w. for harp seals; 0.67 Bq/kg w.w. for the white whale sample; 0.24 Bq/kg w.w. for the blue whale; and below detection limit for the walrus. Significant differences in 137Cs specific activities between some of the species were found. Ringed seals had higher specific activities than the other seal species in the study. Bearded seals and hooded seals had similar values, which were both significantly lower than the harp seal values. The results in the present study are consistent with previous reported results, indicating low specific activities of 137Cs in Arctic marine mammals in the Barents Sea and Greenland Sea region during the last 20 years. The species specific differences found may be explained by varying diet or movement and distribution patterns between species. No age related patterns were found in specific activities for the two species (polar bears and hooded seals) for which sufficient data was available. Concentration factors (CF) of 137Cs from seawater were determined for polar bears, ringed, bearded, harp and hooded seals. Mean CF values ranged from 79+/-32 (SD) for bearded seals sampled in 2002 to 244+/-36 (SD) for ringed seals sampled in 2003 these CF values are higher than those reported for fish and benthic organisms in the literature, suggesting bioaccumulation of 137Cs in the marine ecosystem.

Age Factors↗
PubMed2001

Polycyclic aromatic hydrocarbons (PaHs) in subcutaneous biopsies of Mediterranean cetaceans.

The aim of the present study was to measure polycyclic aromatic hydrocarbon (PAH) levels in free-ranging Mediterranean cetaceans as they are likely to cause chemical stress in the organisms of this basin. Blubber samples were collected from live specimens of fin whales (Balaenoptera physalus) and striped dolphins (Stenella coeruleoalba) by means of biopsies, a non-destructive biological method. Fin whales were sampled in the Ligurian Sea, whereas striped dolphins were collected in the Ligurian and the Ionian Seas. A fingerprint of 14 PAHs was obtained for both species. In whales, the median value of total PAHs was 1970 ppb fresh weight (f.w.) while median carcinogenic PAH values were 89.80 ppb f.w.; in dolphins, the median values of total and carcinogenic PAHs were 29,500 and 676.00 ppb f.w., respectively. The different PAH values between the two species can be attributed to the different positions they take in the Mediterranean food web. The sampling period significantly influenced PAH concentrations of fin whales.

Adipose Tissue↗
PubMed1999

Complete nucleotide sequence of pig (Sus scrofa) mitochondrial genome and dating evolutionary divergence within Artiodactyla.

The complete nucleotide sequence of the pig (Sus scrofa) mitochondrial genome, containing 16613bp, is presented in this report. The genome is not a specific length because of the presence of the variable numbers of tandem repeats, 5'-CGTGCGTACA in the displacement loop (D-loop). Genes responsible for 12S and 16S rRNAs, 22 tRNAs, and 13 protein-coding regions are found. The genome carries very few intergenic nucleotides with several instances of overlap between protein-coding or tRNA genes, except in the D-loop region. For evaluating the possible evolutionary relationships between Artiodactyla and Cetacea, the nucleotide substitutions and amino acid sequences of 13 protein-coding genes were aligned by pairwise comparisons of the pig, cow, and fin whale. By comparing these sequences, we suggest that there is a closer relationship between the pig and cow than that between either of these species and fin whale. In addition, the accumulation of transversions and gaps in pig 12S and 16S rRNA genes was compared with that in other eutherian species, including cow, fin whale, human, horse, and harbor seal. The results also reveal a close phylogenetic relationship between pig and cow, as compared to fin whale and others. Thus, according to the sequence differences of mitochondrial rRNA genes in eutherian species, the evolutionary separation of pig and cow occurred about 53-60 million years ago.

Amino Acids↗
PubMed1999

Biomarkers of DNA damage in marine mammals.

Certain environmental contaminants found in marine mammals have been shown to cause DNA damage and cancer. The micronuclei (MN), sister chromatid exchange (SCE) and/or chromosome aberration (CA) assays were used to assess baseline (spontaneous) levels of DNA damage in blood lymphocytes of individuals of the relatively healthy and lightly contaminated Arctic beluga whale (Delphinapterus leucas), Sarasota Bay, FL, bottlenose dolphin (Tursiops truncatus) and Northwestern Atlantic grey (Halichoerus grypus) and harp (Phoca groenlandicus) seal populations. MN cell (MNC) frequencies ranged between 2 and 14/1000 binucleated (BN) cells and were statistically similar between species. In bottlenose dolphins, MNC frequency was correlated with age and was significantly higher in females than in males. No intraspecific variation in MNC frequency was found in beluga whales. Intraspecific variation was not tested in seals due to the small sample size. Frequencies of SCEs and total CAs, excluding gaps, ranged, respectively, between 1 and 15 SCE(s)/per cell and 4-6 CAs/100 cells in beluga whales. SCE and CA frequencies did not vary with age or sex in beluga whales. The MN, SCE and CA assays were found to be practical tools for the detection of DNA damage in marine mammals and could be used in the future to compare DNA damage between relatively lightly and highly contaminated populations.

Animals↗
PubMed2004

Congener-specific accumulation and food chain transfer of polybrominated diphenyl ethers in two arctic food chains.

Congener-specific accumulation and prey to predator transfer of 22 polybrominated diphenyl ethers (PBDEs) were assessed in polar cod, ringed seal, polar bear, and beluga whale. Although the concentrations found were relatively low, these results show that PBDEs have reached the Arctic. PBDE congeners 47, 99, and 100 were dominant in all species studied. The pattern in ringed seal was somewhat simpler than in polar cod, with PBDE 47 accounting for more than 90% of the total PBDEs. In contrast, beluga whales, feeding on prey similar to that of ringed seals, showed higher PBDE levels and a more complex PBDE pattern than ringed seals. In contrast, polar bears contained only PBDE 47 in relatively small amounts. These differences in levels and patterns are likely due to species-specific differences in PBDE metabolism and accumulation. The metabolic index suggested that PBDEs 47 and 99 accumulate to the same magnitude as PCB 153 (PCB = polychlorinated biphenyl) in ringed seals and beluga whales. In contrast to beluga whales, ringed seals can metabolize PBDE 100 to some extent. Polar bears are seemingly capable of metabolizing virtually all PBDEs and are therefore unsuitable as indicators for PBDE contamination in the environment.

Animals↗
PubMed2003

Perfluorinated chemicals infiltrate ocean waters: link between exposure levels and stable isotope ratios in marine mammals.

This is the first study to report on concentrations of perfluorinated organochemicals (FOCs) in marine mammals stranded along the southern North Sea coast in relation to stable nitrogen and carbon isotope ratios (delta15N and delta13C). The presence of FOCs in top predators such as marine mammals would indicate a potential biomagnification of these compounds and their widespread occurrence. Liver and kidney tissues of nine marine mammal species have been sampled. Among all the measured FOCs compounds, PFOS (perfluorooctane sulfonate) was predominant in terms of concentration. The highest PFOS concentrations were found in the liver of harbor seal compared to white-beaked dolphin, harbor porpoise, gray seal, sperm whale, white-sided dolphin, striped dolphin, fin whale, and hooded seal. PFOS concentrations differed significantly between sexes and age classes in harbor porpoises. Stable isotope measurements (delta13C and delta15N) were used in this study to describe the behavior of contaminants in food webs. We found a significant (p < 0.05) linear relationship between PFOS concentrations in livers of harbor porpoises and both muscle delta13C and delta15N measurements. Harbor and gray seals and white-beaked dolphin, which displayed the highest trophic position, contained the highest PFOS levels, while offshore feeders such as sperm whales, fin whales, striped dolphin, and white-sided dolphin showed lower PFOS concentrations than inshore species.

Age Factors↗
PubMed2002

Selective habituation shapes acoustic predator recognition in harbour seals.

Predation is a major force in shaping the behaviour of animals, so that precise identification of predators will confer substantial selective advantages on animals that serve as food to others. Because experience with a predator can be lethal, early researchers studying birds suggested that predator recognition does not require learning. However, a predator image that can be modified by learning and experience will be advantageous in situations where cues associated with the predator are highly variable or change over time. In this study, we investigated the response of harbour seals (Phoca vitulina) to the underwater calls of different populations of killer whales (Orcinus orca). We found that the seals responded strongly to the calls of mammal-eating killer whales and unfamiliar fish-eating killer whales but not to the familiar calls of the local fish-eating population. This demonstrates that wild harbour seals are capable of complex acoustic discrimination and that they modify their predator image by selectively habituating to the calls of harmless killer whales. Fear in these animals is therefore focused on local threats by learning and experience.

Acoustic Stimulation↗
PubMed1984

Anticoagulant properties of heparin preparations from different animal sources with equivalent high affinity for antithrombin III.

Three kinds of heparins from different animal sources--hog-intestinal heparin (167 USP units/mg), bovine-lung heparin (149 USP units/mg) and whale-intestinal heparin (Balaenoptera borealis L., 174 USP units/mg)--were fractionated through an antithrombin III-Sepharose column and high-affinity (HA) subfractions (1.2-1.8 M NaCl eluted fractions) were separated. These HA heparin specimens, which were confirmed to be equivalent in strength of binding to antithrombin III, were compared with respect to the anticoagulant properties by means of [1] whole blood clotting assay and [2] thrombin or [3] factor Xa inhibition assay. The activities of whale HA heparin were [1] 82%, [2] 38%, and [3] 45%, respectively, of those of the hog and bovine HA heparins (which showed approximately identical activities). Since it was confirmed that the affinity for antithrombin III of whale heparin is of the same nature as those of hog and bovine heparins by (a) fluorometric titration and (b) affinity chromatography on an antithrombin III-Sepharose column saturated with hog HA heparin, the above difference in anticoagulant properties between the whale HA heparin and hog and bovine HA heparins may be considered to be due to a difference in undescribed saccharide-sequence structures at sites other than the well-characterized antithrombin III-binding sites, suggesting strongly that some heparin saccharide structures other than antithrombin III-binding oligosaccharide participate in the anticoagulant activity dependent on antithrombin III-heparin binding--at least in the manifestation of anti-thrombin activity and the enhancement of anti-Xa activity.

Animals↗
PubMed2004

Localization of marine mammals near Hawaii using an acoustic propagation model.

Humpback whale songs were recorded on six widely spaced receivers of the Pacific Missile Range Facility (PMRF) hydrophone network near Hawaii during March of 2001. These recordings were used to test a new approach to localizing the whales that exploits the time-difference of arrival (time lag) of their calls as measured between receiver pairs in the PMRF network. The usual technique for estimating source position uses the intersection of hyperbolic curves of constant time lag, but a drawback of this approach is its assumption of a constant wave speed and straight-line propagation to associate acoustic travel time with range. In contrast to hyperbolic fixing, the algorithm described here uses an acoustic propagation model to account for waveguide and multipath effects when estimating travel time from hypothesized source positions. A comparison between predicted and measured time lags forms an ambiguity surface, or visual representation of the most probable whale position in a horizontal plane around the array. This is an important benefit because it allows for automated peak extraction to provide a location estimate. Examples of whale localizations using real and simulated data in algorithms of increasing complexity are provided.

Algorithms↗
PubMed2004

Distributive and phagocytic characteristics of hepatic macrophages in five cetaceans belonging to Delphinidae and Ziphiidae.

Details of morphology and distribution of hepatic macrophages in cetaceans were investigated using the immunohistochemistry with an antibody (SRA-E5) generated against human macrophage scavenger receptor antigen. Liver samples were obtained from five species of cetaceans (Baird's beaked whales, short-finned pilot whales, Risso's dolphins, bottlenose dolphins, and pantropical spotted dolphins). Except for two species of whales, the number of SRA-E5-positive Kupffer cells was greatest in the perivenous zone (zone 3), followed by the mid-zonal (zone 2) and periportal (zone 1) zones; this distribution pattern was different from that in cattle examined here and previously reported rodents with the highest number in zone 1. The frequency of Kupffer cell in each of zones was significantly different among species, and interestingly, the total mean of the Kupffer cell number in three zones increased as the body-length of species was small. In cetaceans, Kupffer cells in zone 1 appeared larger and more stellate in shape, whereas those in zone 3 were smaller and rounder. All cetaceans but Baird's beaked whales had the black pigment-containing Kupffer cells, with the greatest number in zone 3, and macrophages with the similar pigments were also seen in the hepatic intermediate septa, indicating an active phagocytosis. Most of the black pigments were considered to be lipofuscin and such pigments were not seen in the bovine livers. These results indicate that cetacean hepatic macrophages show differences in the distribution and phagocytosis among hepatic lobular zones, or between cetacean species and terrestrial animals.

Animals↗
PubMed1990

Effects of different heparins on the enhancement of the thrombin-antithrombin reaction.

Kinetic analyses were made of the thrombin/antithrombin III (ATIII) reaction in the presence of catalytic amounts of three kinds of mammalian HA-heparin (bovine, pig and whale), which have the same affinities for ATIII. In the absence of NaCl, the first-order rate constant was higher with whale HA-heparin than with the other two HA-heparins. However, the strength of the interactions of thrombin with the HA-heparins was in the order, bovine greater than pig greater than whale. Thus, the slow reactions in the case of bovine and pig HA-heparins were probably due to preferential binding of the two HA-heparins to thrombin rather than to ATIII, thus causing the HA-heparins to be inhibitory for the reaction by reducing the turnover rates of the polysaccharides as catalysts. In the presence of 0.15M NaCl, the first-order rate constants were slightly higher in the case of pig HA-heparin than of the other two HA-heparins. Since the strength of the interactions of these HA-heparins with thrombin was in the order, pig greater than or equal to bovine greater than whale, the faster reactions were probably due to higher associations of the enzyme with the essential HA-heparin-ATIII complex.

Animals↗
PubMed2006

Competition among penguins and cetaceans reveals trophic cascades in the western Ross Sea, Antarctica.

An apparent trophic cascade that appears during summer in the western Ross Sea, Antarctica, explains why the Antarctic silverfish (Pleuragramma antarcticum) there becomes cannibalistic; its principal prey, crystal krill (Euphausia crystallorophias) becomes scarce; and the diatom community is minimally grazed compared to adjacent areas. The krill is the major grazer of diatoms. On the basis of fieldwork at Ross Island, we suggest that the cascade results from foraging by unusually numerous Adélie Penguins (Pygoscelis adeliae), minke whales (Balaenoptera bonaerensis), and fish-eating killer whales (Orcinus orca). These species and other top predators apparently deplete the krill and silverfish. In drawing our conclusions, we were aided by two "natural experiments." In one "experiment," large, grounded icebergs altered the seasonal pattern of change in regional sea-ice cover, but not the seasonal change in penguin diet and foraging behavior that was also detected during the pre-iceberg era. In the other "experiment," a short-term polynya (opening in the ice) brought penguins and whales together in a confined area, this time altering both penguin diet and foraging behavior. We conclude that the foraging of penguins and whales, and not a formerly hypothesized seasonal decrease in sea-ice cover, explains (1) the annual switch in the penguins' prey from krill to silverfish, (2) the subsequent lengthening of penguin foraging trips, and (3) a marked decline of cetaceans in the area later in the season. Reduction in the middle-trophic-level prey is expressed in the relaxed grazing pressure on phytoplankton.

Animals↗
PubMed1990

Transient spectroscopy of the reaction of cyanide with ferrous myoglobin. Effect of distal side residues.

The reaction of cyanide metmyoglobin with dithionite conforms to a two-step sequential mechanism with formation of an unstable intermediate, identified as cyanide bound ferrous myoglobin. This reaction was investigated by stopped-flow time resolved spectroscopy using different myoglobins, i.e. those from horse heart, Aplysia limacina buccal muscle, and three recombinant derivatives of sperm whale skeletal muscle myoglobin (Mb) (the wild type and two mutants). The myoglobins from horse and sperm whale (wild type) have in the distal position (E7) a histidyl residue, which is missing in A. limacina Mb as well as the two sperm whale mutants (E7 His----Gly and E7 His----Val). All these proteins in the reduced form display an extremely low affinity for cyanide at pH less than 10. The differences in spectroscopy and kinetics of the ferrous cyanide complex of these myoglobins indicate a role of the distal pocket on the properties of the complex. The two mutants of sperm whale Mb are characterized by a rate constant for the decay of the unstable intermediate much faster than that of the wild type, at all pH values explored. Therefore, we envisage a specific role of the distal His (E7) in controlling the rate of cyanide dissociation and also find that this effect depends on the protonation of a single ionizable group, with pK = 7.2, attributed to the E7 imidazole ring. The results on A. limacina Mb, which displays the slowest rate of cyanide dissociation, suggests that a considerable stabilizing effect can be exerted by Arg E10 which, according to Bolognesi et al. (Bolognesi, M., Coda, A., Frigerio, F., Gatti, C., Ascenzi, P., and Brunori, M. (1990) J. Mol. Biol. 213, 621-625), interacts inside the pocket with fluoride bound to the ferric heme iron. A mechanism of control for the rate of dissociation of cyanide from ferrous myoglobin, involving protonation of the bound anion, is discussed.

Animals↗
PubMed1989

Cetacean relaxin. Isolation and sequence of relaxins from Balaenoptera acutorostrata and Balaenoptera edeni.

The tendency toward extremely high variability among relaxins derived from purportedly closely related species has come to an abrupt end with the discovery of quasi-porcine relaxin in the minke whale (Balaenoptera acutorostrata) and the Bryde's whale (Balaenoptera edeni). An aqueous abstract of the corpora lutea of the two baleen whales contained significant amounts of relaxin-like activity as determined by a mouse bioassay and by cross-reactivity with anti-pig relaxin antibodies. The activity could be isolated and purified to homogeneity. Sequence analysis revealed that both whale relaxins differed from each other by about 3 residues, whereas the relaxin of B. edeni differed at only one position from that of pig relaxin. The similarity appears to include even the chain length heterogeneity observed at the C-terminal end of the B chain in porcine relaxin which is produced by a peculiar mode of connecting peptide removal from the pro-hormone. This finding may well represent one of the better documented challenges to the current paradigm of molecular evolution.

Amino Acid Sequence↗
PubMed1978

Prevalence of vesicular exanthema of swine antibodies among feral mammals associated with the southern California coastal zones.

Serum-neutralizing antibodies to both vesicular exanthema of swine virus (VESV) and San Miguel sea lion virus (SMSV) were found in a number of animal species having an association with the southern California coastal zones. California sea lions (Zalophus californianus) had antibodies to 9 VESV types (A48, C52, D53, E54, F54, G55, I55, J56, and K56). Fur seals (Callorhinus ursinus) and elephant seal pups (Mirounga angustirostris) were tested for antibodies to 6 VESV types and all were negative. California gray whales (Eschrichtius robustus) were tested for antibodies to 9 VESV types, and sperm whale (Physeter catodon), finback whale (Balaenoptera physalus), and sei whale (Balaenoptera borealis) were tested for 6 VESV types. Among the last 4 species, antibodies were present for each VESV type except C52, and all species were positive for antibodies to 2 or more VESV types. Feral swine from both Santa Cruz Island and Santa Catalina Island were tested and antibodies were present for 8 of 9 VESV types and all SMSV types except SMSV-4. One donkey from San Miguel Island was positive for VESV I55 and 2 were positive for SMSV-2.

Animals↗