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Vestimentiferan on a whale fall.

Discovery of chemosynthetic communities associated with whale bones led to the hypothesis that whale falls may serve as stepping-stones for faunal dispersal between disjunct hydrothermal vents and cold seeps on the ocean floor (1). The initial observation was followed by a faunal inventory that revealed a diverse assemblage of microbes and invertebrates, supported by chemoautotrophic production, living in close proximity to whale remains (2, 3). To date, the conspicuous absence from whale falls of vestimentiferan tubeworms (a predominant constituent of eastern Pacific vent and seep habitats) has been a major objection to the stepping-stone hypothesis (4-5). We report the first evidence of a vestimentiferan tubeworm associated with a whale fall (Fig. 1). The tubeworm, Escarpia spicata, was identified by morphological criteria and DNA sequence data from a portion of the mitochondrial cytochrome oxidase C subunit I (COI) gene. Additionally, the bacterial endosymbiont in the tubeworm possessed a 16S rRNA gene that was similar to that of endosymbionts from vestimentiferans in sedimented cold-seep environments.

Animals↗

Serology and virology of the bowhead whale (Balaena mysticetus L.).

Sera from four bowhead whales (Balaena mysticetus L.) were examined for the presence of specific antibodies, and tissue and swab samples from six and four animals respectively were processed for isolation of viruses and for initiation of bowhead whale cell cultures. All sera were negative for antibodies to nine serovars of Leptospira interrogans and to 21 orthomyxovirus subtypes and a paramyxovirus (Newcastle disease virus). All sera were positive, however, for neutralizing antibodies to one or more calicivirus serotypes. Two untyped adenoviruses were isolated from colon samples of two different whales, but neutralizing antibodies to the agents could not be demonstrated in any sera. Three primary bowhead whale cell cultures were derived from kidney (two cultures) and testis (one culture), from three individual whales.

Animals↗

Bone fluoride concentrations in beluga whales from Canada.

Beluga whales (Delphinapterus leucas) from the St. Lawrence Estuary have been reported to have dental and bone abnormalities. To determine whether these lesions could be caused by high exposure to fluorides, we measured bone fluoride levels in eight beluga whales stranded on the shores of the St. Lawrence Estuary (Quebec, Canada), and in nine beluga whales killed by Inuit hunters in the Hudson Bay (North Western Territories, Canada). In both groups, fluoride concentrations were higher than those found in terrestrial mammals intoxicated by fluorides. Unexpectedly, fluoride concentration was significantly higher in beluga whales from the Hudson Bay (mean +/- SD: 10.365 +/- 1.098 ppm) than in beluga whales from the St. Lawrence Estuary (4.539 +/- 875 ppm) and was positively correlated with age in the latter population. Differences in diet might explain the differences in fluoride concentrations found between these two populations.

Aging↗

Photoreceptor ultrastructure of the amphipod, Cyamus ceti (Linné, 1758), an ectoparasite of bowhead, right and gray whales.

Light and electron microscopic findings are presented for the first time on a species of amphipod whale louse, Cyamus ceti, with reference to the paired photoreceptor structures of the head. Whale lice are crustacean ectoparasites of large, slow moving whales. Twenty-two known species parasitize mainly baleens but also toothed whales. Samples were collected from Alaskan bowhead whales during an Eskimo hunt. The photoreceptors, or compound eyes, were sessile and located on the dorsal cephalon. Each photoreceptor contained approximately 50 visual ommatidial units. The general organization was similar to other amphipod crustacean compound eyes. The single ommatidial unit consisted of 1) an overlying cuticle, 2) crystalline cone secreted by cone cells, 3) a fusiform layered rhabdom, 4) surrounding pigmented retinular cells, and 5) basal lamina and axon. The cuticle over each eye was translucent, convex, and thinner than the rest of the cranial cuticle. The photosensitive rhabdom contained a central core of alternating microvillous projections, and the rhabdomere unit was formed by five or six retinular cells. There were numerous electron-dense and electron-lucent granules within the cytoplasm of the retinular cells, corresponding to visual pigment or reflecting granules. The eyes most likely play a role in detecting light direction, duration and/or intensity, which may direct molting, reproduction, or other functions of this cetacean ectoparasite.

Animals↗

Thyroid hormone balance in beluga whales, Delphinapterus leucas: dynamics after capture and influence of thyrotropin.

Ten beluga whales, Delphinapterus leucas, were captured in the Churchill River, Manitoba, held for up to five days, and then released. Blood samples were obtained immediately after capture and at 6-7 h intervals thereafter to monitor changes in circulating levels of thyroid hormones (TH). In six of the whales, total and free thyroxine (T4) and triiodothyronine (T3) declined steadily, whereas reverse-T3 (rT3) showed a transient increase during the first 24-36 h, followed by a decrease to below initial values. The changes in TH may have been due to glucocorticoid-mediated reduction in endogenous thyroid stimulating hormone (TSH), and inhibition of 5'-monodeiodinase in peripheral tissue. Two whales were given 10 IU of bovine TSH immediately after capture, and again one and two days later, resulting in successive increases in all TH, which remained elevated for at least 24 h after the last injection. Thereafter, circulating levels declined as in the untreated whales. Two whales receiving a single TSH injection on the fourth day responded with an increase in plasma TH comparable to that observed following the first TSH injection in the other two animals. Average (+/- SD) circulating level of rT3 at capture was 6.3 +/- 3.1 nmol/L, which is higher than reported for any other mammal and was significantly correlated with the naturally elevated levels of T4 that occur in belugas occupying estuaries during the summer.

Animals↗

Developmental changes of the fore- and hind-limbs in the fetuses of the southern minke whale, Balaenoptera acutorostrata.

15 fetuses of southern minke whale, Balaenoptera acutorostrata (from 8.2 mm C.R.L. to 38.9 mm C.R.L.), obtained from the Kyodo whale company in Japan (in whaling seasons from 1982 to 1986), were used for the present study. The fore-limb first appears in the 11.4 mm fetus, and the hind-limb in the 15.3 mm fetus. The fore-limb develops progressively during gestation, while the hind-limb disappears by the 38.9 mm fetus. The critical period of limb development in southern minke whale was shown to be at a stage between that in the small sized species; Common Porpoise, Phocoena communis and Striped Dolphin, Prodelphinus caeruleoalbus, and in large one; Humpback Whale, Megaptera nodusa.

Animals↗

Genetic impact of an unusual group mortality among humpback whales.

Mass mortalities, due to infectious disease or toxic algal blooms, are known to have severe demographic impacts on marine mammal populations. The genetic impacts of these events, however, have received little attention. To investigate the genetic consequences of an unusual group mortality among humpback whales, we compared the mitochondrial DNA haplotypes of 10 whales poisoned by mackerel contaminated with a dinoflagellate neurotoxin to those of 32 live whales from the same regional population. Two haplotypes that were rare in the reference sample of live whales accounted for eight of the 10 poisoned whales. A randomized test of independence, based on 500 permutations of the data matrix, showed significant differences in the frequencies of haplotypes in the two samples (P < .002). This is the first demonstration that group mortality events in marine mammals can have unpredictable genetic consequences and points to a need to evaluate ecological disasters within the context of the genetic mosaic of natural populations.

Animals↗

Bioaccumulation of naturally occurring mixed halogenated dimethylbipyrroles in whale and dolphin products on the Japanese market.

Mixed halogenated dimethyl bipyrroles (HDBPs), which are thought to be produced naturally, were quantified in whale and dolphin products marketed for human consumption in Japan. The major component of HDBPs was 3,3',4,4'-tetrabromo-5,5'-dichloro-1,1'-dimethyl -2,2'-bipyrrole (Br(4)Cl(2)-DBP), accounting for 85% of the total of five HDBPs detected, followed by Br(3)Cl(2)-DBP. Mean concentrations of HDBPs ranged from 0.27 microg/g lipid (n = 31) in minke whale (Balaenoptera acutorostrata) from the northwest Pacific Ocean to 11.8 microg/g lipid (n = 33) in bottlenose dolphin (Tursiops trucatus) from the southwest Japanese coastal water. At higher levels, HDBPs made up 37% of the total organohalogen body burden in Dall's porpoise (Phocoenoides dalli), whereas the contribution was less than 8.9% in minke whales. In two data subsets from Baird's beaked whale (Berardius bairdii), the products from the Pacific Ocean contained significantly higher concentrations of HDBPs than those from the Sea of Japan. Furthermore, the geographical distribution of HDBPs did not resemble those of ubiquitous anthropogenic organochlorines, such as polychlorinated biphenyl (PCBs). Higher concentration ratios of SigmaHDBP/SigmaPCB and different patterns of HDBP congeners were observed in whale products from the Asia-Pacific as compared to non-Pacific Ocean mammals reported previously. These results support the hypothesis that HDBPs and anthropogenic organochlorines have different sources and that the consumption of HDBPs by Japanese individuals could be an exposure/health risk.

Animals↗

Diurnal and annual changes in serum cortisol concentrations in Indo-Pacific bottlenose dolphins Tursiops aduncus and killer whales Orcinus orca.

Until present, fundamental studies on cortisol secretory patterns have not been conducted in cetaceans. The objectives of this study were: (1) to examine diurnal changes in serum cortisol concentrations in Indo-Pacific bottlenose dolphins Tursiops aduncus and killer whales Orcinus orca, (2) to investigate annual cortisol changes in killer whales, and (3) to investigate the relationship between cortisol and sex steroids (testosterone and progesterone) concentrations in killer whales. Diurnal changes in serum cortisol concentrations were investigated at various intervals in the two species. In Indo-Pacific bottlenose dolphins, serum cortisol levels exhibited the same episodic fluctuations for 24 h as did diurnal terrestrial mammals: cortisol levels were lower at 18:00 h and higher in the early morning. In killer whales, cortisol concentrations continued to decrease until 18:00 h, after which they fluctuated, and then increased in the next morning. Annual changes in cortisol levels were investigated by collecting blood samples every two weeks from two male killer whales and a pregnant female one twice per day (during 09:00-10:00 and 16:00-17:00 h) throughout a one-year period. Regarding sera collected during 09:00-10:00 h from the female, cortisol concentrations showed cyclic changes having about 4-month intervals. In males, cortisol showed higher concentrations in winter and lower concentrations during the summer season. There was a negative correlation between cortisol and progesterone levels in the female and a negative correlation was also observed between cortisol and testosterone in male no. 2. In the female and male no. 1, cortisol levels during 09:00-10:00 h were significantly higher than those during 16:00-17:00 h, and their data are considered to support observations regarding diurnal changes in cortisol levels in the two cetacean species.

Animals↗

Expression and functional characterization of killer whale (Orcinus orca) interleukin-6 (IL-6) and development of a competitive immunoassay.

Interleukin-6 (IL-6) is a cytokine that can reach detectable systemic levels and is a major inducer of the acute phase response. As such, clinical assays to identify this cytokine in mammalian sera are of diagnostic value. A 558 base-pair (bp) fragment of killer whale IL-6 was cloned and expressed as a 21 kDa protein in Escherichia coli. Biological activity of the recombinant killer whale IL-6 (rkwIL-6) was demonstrated using the IL-6-dependent B9 mouse hybridoma cell line; acute phase sera from a killer whale and supernatants from lipopolysaccharide (LPS)-stimulated killer whale peripheral blood mononuclear cells (PBMCs) also supported the proliferation of the B9 hybridoma. Rat anti-mouse IL-6 receptor antibody effectively blocked biological activity of all three sources of IL-6. Polyclonal antisera, specific for the recombinant protein, were obtained by successive immunization of a rabbit with rkwIL-6. The polyclonal antibody was capable of neutralizing the biological activity of both recombinant and native kwIL-6. A competitive enzyme-linked immunosorbent assay (ELISA) was developed using the polyclonal rabbit anti-rkwIL-6 and the recombinant protein; sensitivity of the assay was in the range of 1 ng/ml. The ELISA was subsequently used to identify the presence of native IL-6 in acute phase sera of two species of delphinidae, a killer whale and a bottlenose dolphin. The application of quantitative cytokine assays as diagnostic tools for monitoring cetacean health are becoming feasible as many animals are now being trained for fluke presentation, making blood collection a routine procedure.

Animals↗

Persistent organochlorines in minke whale (Balaenoptera acutorostrata) and their prey species from the Antarctic and the North Pacific.

Persistent organochlorines such as polychlorinated biphenyls (PCBs), DDTs, chlordane compounds (CHLs), hexachlorocyclohexanes (HCHs) and hexachlorobenzene (HCB) were determined in the blubber of minke whale and its diet collected from the Antarctic and the North Pacific Oceans. Residue levels of these compounds (except HCB) in minke whale from the Antarctic were apparently lower than those from the North Pacific. This is due to the lower levels of these pollutants in the southern hemisphere than in the northern hemisphere and the specific feeding habit of the minke whale from the Antarctic which feeds on lower trophic organisms, primarily euphausiids. The north-south difference for HCB residue levels was small, reflecting its dispersible nature through long-range atmospheric transport. Compositions of DDT and CHL compounds in minke whale from the Antarctic were similar to those from the North Pacific. However, the composition of HCH isomers was different between the Antarctic and the North Pacific as was observed in their diet, suggesting a larger or on-going usage of lindane in the southern hemisphere countries. In minke whale from the Antarctic, the elevated level of PCBs residues was noted during a period of 1984 to 1993, implying a continuous discharge of PCBs in the southern hemisphere. A similar discharge was also suggested in the North Pacific, while a decreasing contamination by DDTs was apparent.

Journal Article↗

Population structure of North American beluga whales (Delphinapterus leucas) based on nuclear DNA microsatellite variation and contrasted with the population structure revealed by mitochondrial DNA variation

Beluga whales (Delphinapterus leucas) in North American waters migrate seasonally between wintering areas in broken pack ice and summering locations in estuaries and other open water areas in the Arctic and sub-Arctic. Results from our previous investigation of beluga whale mitochondrial DNA (mtDNA) revealed genetic heterogeneity among beluga from different summering locations that was interpreted as representing a high degree of summering site philopatry. However, mtDNA is maternally inherited and does not reflect mating that may occur among beluga from different summering locations in wintering areas or during annual migrations. To test the possibility that breeding occurs among beluga from different summering locations, genetic variability at five nuclear DNA (nDNA) microsatellite loci was examined in the same animals tested in the mtDNA study. Beluga samples (n = 640) were collected between 1984 and 1994 from 24 sites across North America, mostly during the summer. Whales from the various sites were categorized into eight summering locations as identified by mtDNA analysis, as well as four hypothesized wintering areas: Bering Sea, Hudson Strait (Hudson Strait, Labrador Sea, southwest Davis Strait), Baffin Bay (North Water, east Davis Strait), and St Lawrence River. Microsatellite allele frequencies indicated genetic homogeneity among animals from summering sites believed to winter together but differentiation among whales from some of the wintering areas. In particular, beluga from western North America (Bering Sea) were clearly distinguished from beluga from eastern North America (Hudson Strait, Baffin Bay, and St Lawrence River). Based upon the combined data set, the population of North American beluga whales was divided into two evolutionarily significant units. However, the population may be further subdivided into management units to reflect distinct groups of beluga at summering locations.

Journal Article↗

Echolocation signals of free-ranging killer whales (Orcinus orca) and modeling of foraging for chinook salmon (Oncorhynchus tshawytscha).

Fish-eating "resident"-type killer whales (Orcinus orca) that frequent the coastal waters off northeastern Vancouver Island, Canada have a strong preference for chinook salmon (Oncorhynchus tshawytscha). The whales in this region often forage along steep cliffs that extend into the water, echolocating their prey. Echolocation signals of resident killer whales were measured with a four-hydrophone symmetrical star array and the signals were simultaneously digitized at a sample rate of 500 kHz using a lunch-box PC. A portable VCR recorded the images from an underwater camera located adjacent to the array center. Only signals emanating from close to the beam axis (1185 total) were chosen for a detailed analysis. Killer whales project very broadband echolocation signals (Q equal 0.9 to 1.4) that tend to have bimodal frequency structure. Ninety-seven percent of the signals had center frequencies between 45 and 80 kHz with bandwidths between 35 and 50 kHz. The peak-to-peak source level of the echolocation signals decreased as a function of the one-way transmission loss to the array. Source levels varied between 195 and 224 dB re: 1 microPa. Using a model of target strength for chinook salmon, the echo levels from the echolocation signals are estimated for different horizontal ranges between a whale and a salmon. At a horizontal range of 100 m, the echo level should exceed an Orcinus hearing threshold at 50 kHz by over 29 dB and should be greater than sea state 4 noise by at least 9 dB. In moderately heavy rain conditions, the detection range will be reduced substantially and the echo level at a horizontal range of 40 m would be close to the level of the rain noise.

Animals↗

Killer whale (Orcinus orca) hearing: auditory brainstem response and behavioral audiograms.

Killer whale (Orcinus orca) audiograms were measured using behavioral responses and auditory evoked potentials (AEPs) from two trained adult females. The mean auditory brainstem response (ABR) audiogram to tones between 1 and 100 kHz was 12 dB (re 1 mu Pa) less sensitive than behavioral audiograms from the same individuals (+/- 8 dB). The ABR and behavioral audiogram curves had shapes that were generally consistent and had the best threshold agreement (5 dB) in the most sensitive range 18-42 kHz, and the least (22 dB) at higher frequencies 60-100 kHz. The most sensitive frequency in the mean Orcinus audiogram was 20 kHz (36 dB), a frequency lower than many other odontocetes, but one that matches peak spectral energy reported for wild killer whale echolocation clicks. A previously reported audiogram of a male Orcinus had greatest sensitivity in this range (15 kHz, approximately 35 dB). Both whales reliably responded to 100-kHz tones (95 dB), and one whale to a 120-kHz tone, a variation from an earlier reported high-frequency limit of 32 kHz for a male Orcinus. Despite smaller amplitude ABRs than smaller delphinids, the results demonstrated that ABR audiometry can provide a useful suprathreshold estimate of hearing range in toothed whales.

Animals↗

Observations on the external morphology and vasculature of a fetal heart of the bowhead whale, Balaena mysticetus.

BACKGROUND: Specialized demands within the aquatic environment for over some 60 million years have shaped unique morphological expressions in the whales, dolphins, and porpoises (Cetacea). Detailed consideration of these features, particularly in the great whales, has often been constrained by difficulties in securing adequate specimens for study. We had the opportunity to examine external heart morphology in a rarely obtained and prepared specimen from the bowhead whale, Balaena mysticetus. METHODS: The external morphology and in situ relations of a formalin-perfused heart were examined grossly in a near-term bowhead fetus. Latex injections assisted visualization of coronary vasculature. Magnetic resonance imaging was used to clarify heart positioning within the thoracic cavity in two younger (early and mid-gestational) intact fetuses. RESULTS: The heart was globular in form, with a blunt apex and wide base; it was laterally broad relative to height yet considerably compressed between nearly planar atrial (diaphragmatic) and auricular (sternocostal) surfaces. The heart constituted 0.01 of body mass in the near-term fetus. Within the thoracic cavity, the heart tilted forward on its long axis, placing the great basal vessels in the region of the thoracic inlet. The aorta extended forward from mid-base in parallel with the pulmonary trunk, arched sharply to the left, producing in succession the brachiocephalic trunk, left common carotid artery, and the left subclavian artery. Bifurcation of the brachiocephalic trunk yielded the right common carotid and right subclavian arteries. The distal portion of the aortic arch was linked to the pulmonary trunk via the ductus arteriosus. The aorta then swung caudally over the heart base, descending beneath the bodies of the thoracic vertebrae. The ascending aorta featured three bulbous sinuses immediately distal to the three semilunar cusps of the aortic valve. Originating along the distal boundaries of the left and right sinuses were the left and right coronary arteries. The arteries were similar in size and, because each sent contributions along their respective coronary and interventricular grooves, the heart can be described as bilateral relative to arterial supply. Anastomoses were common within and between the two arteries. Venous return from the heart was comprised of the great, middle, and right cardiac veins, all three converging in the coronary sinus. The right cardiac vein also included tributaries that emptied directly into the right atrium. CONCLUSIONS: External heart morphology in the fetal bowhead whale examined was distinguished by a laterally broad conformation with significant compression between its cranial and caudal surfaces. Aortic bulb configuration in combination with an expandable aortic arch may support blood service to the heart during diastole. Vascular service to the heart featured a complex vessel network with extensive intraarterial and intravenous anastomoses that enable many alternate blood perfusion pathways and may be adaptive to water-column-pressure fluctuations experienced by a large diving mammal.

Animals↗

Observations on the muscles of the eye of the bowhead whale, Balaena mysticetus.

The muscles of the eyelids and the extraocular muscles of mysticete whales are poorly described for a variety of reasons, including considerable difficulty in obtaining specimens. Our objective is to provide such a description for the bowhead whale, Balaena mysticetus. This study has examined the gross anatomy of the region in six specimens (five adults, one fetus) of the bowhead whale. Results show that the muscles associated with the eye are well developed and possess several distinctive features. For example, precise limits of each extraocular muscle are difficult to determine along their entire length because these muscles intermingle with one another near their insertion. Furthermore, some fibers from these muscles (except the retractor bulbi) also insert into the eyelids. Pulling on these muscles to simulate contraction results in movement of the eyelids and suggests a role for these muscles in palpebral retraction. Insertion of a large levator palpebrae superioris muscle into the upper eyelid further enhances opening of the palpebral fissure. Another unusual feature is the presence of tunnel-like structures that redirect the dorsal and ventral oblique muscles. The dorsal oblique muscle is redirected caudally about 90 degrees, then directed medially by another 90 degrees. These directional changes are accomplished via a connective tissue tunnel derived in part from the fibrous connective tissue of the dorsal rectus and the levator palpebrae superioris muscles. In most terrestrial mammals, a similar change in direction is accomplished by a cartilaginous trochlea. The ventral oblique muscle originates via a slender tendon from the frontal bone and undergoes a similar radical change in direction. Its tendon of insertion undergoes about a 90-degree change in direction that is accomplished through a tunnel-like structure derived from fibrous connective tissue of the ventral rectus muscle. Based on the morphology of the musculature presented, it is likely that the eyeballs and eyelids of the bowhead whale are quite mobile and appear capable of complex movement. The possibility of retraction and protrusion of the eyeball is discussed.

Age Factors↗

The fibrocartilage skeleton and related structures of the ventral pouch of balaenopterid whales.

The mandibular symphysis of rorqual whales, whales of the genera Megaptera and Balaenoptera, is characterized by a Y-shaped fibrocartilage structure that lies in the substance of the muscular ventral pouch of these animals. The stem of the structure joins with the symphysis and is usually indicated externally by an unfurrowed median strip of blubber that has been called the "cutwater" by earlier writers. The arms of the Y pass back and are superficially indicated in all rorqual whales as a ridge running parallel to the rami of the mandibles. This fibrocartilage skeleton of the pouch is most closely related to the mylohyoid muscle. The function of the fibrocartilage Y is probably linked with the jaw mechanics of these whales, but its precise function is otherwise not known.

Animals↗

Comparison between the complete mtDNA sequences of the blue and the fin whale, two species that can hybridize in nature.

The sequence of the mitochondrial DNA (mtDNA) molecule of the blue whale (Balaenoptera musculus) was determined. The molecule is 16,402 bp long and its organization conforms with that of other eutherian mammals. The molecule was compared with the mtDNA of the congeneric fin whale (B. physalus). It was recently documented that the two species can hybridize and that male offspring are infertile whereas female offspring may be fertile. The present comparison made it possible to determine the degree of mtDNA difference that occurs between two species that are not completely separated by hybridization incompatibility. The difference between the complete mtDNA sequences was 7.4%. Lengths of peptide coding genes were the same in both species. Except for a small portion of the control region, disruption in alignment was usually limited to insertion/deletion of a single nucleotide. Nucleotide differences between peptide coding genes ranged from 7.1 to 10.5%, and difference at the inferred amino acid level was 0.0-7.9%. In the rRNA genes the mean transition difference was 3.8%. This figure is similar in degree to the difference (3.4%) between the 12S rRNA gene of humans and the chimpanzee. The mtDNA differences between the two whale species, involving both peptide coding and rRNA genes, suggest an evolutionary separation of > or = 5 million years. Although hybridization between more distantly related mammalian species may not be excluded, it is probable that the blue and fin whales are nearly as different in their mtDNA sequences as hybridizing mammal species may be.

Adenosine Triphosphatases↗