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Genetic variation in Phoca vitulina (the harbour seal) revealed by DNA fingerprinting and RAPDs.

Genetic variation in two harbour seal (Phoca vitulina) populations from the Dutch Wadden Sea and Scotland was examined by RAPD analysis and DNA fingerprinting. For comparison a population of grey seals (Halichoerus grypus) was studied. The RAPD method revealed a very low number of polymorphic bands. The multilocus DNA fingerprinting technique showed very low levels of variation both within and between populations of the harbour seals. For one probe a significant difference in variation between populations was demonstrated. The estimated average heterozygosity, however, diverged more strongly between the two harbour seal populations because of more frequent occurrence of rare alleles in the Scottish population. Low levels of genetic variation seem to be characteristic for the subspecies Phoca vitulina vitulina, but not for all marine mammals in the North Sea, as the grey seal population showed significantly higher variation.

Animals↗

Atopobacter phocae gen. nov., sp. nov., a novel bacterium isolated from common seals.

Two strains of a Gram-positive, catalase-negative, facultatively anaerobic, rod-shaped bacterium isolated from common seals were characterized using phenotypic and molecular taxonomic methods. The two strains closely resembled each other based on their biochemical characteristics, and PAGE analysis of whole-cell protein patterns confirmed their close phenotypic affinity. 16S rRNA gene sequencing showed that the two strains were genetically highly related (99.8% sequence similarity) and that they constitute a new line of descent within the lactic acid group of bacteria. The nearest phylogenetic neighbours of the unknown bacterium were Granulicatella spp., with related taxa such as enterococci, carnobacteria, Desemzia incerta, Lactosphaera pasteurii, Melissococcus plutonius, tetragenococci and vagococci more distantly related. Based on phylogenetic and phenotypic evidence it is proposed that the unknown bacterium from seals be classified in a new genus as Atopobacter phocae gen. nov., sp. nov. The type strain of Atopobacter phocae is CCUG 42358T (= CIP 106392T).

Animals↗

Functional relationship between muscles of mastication and the skull with enlarged orbit in the Baikal seal (Phoca sibirica).

We expected that the enlarged area of zygomatic arch, one of some skull characters adapted for enlarged eye, has an influence on form and function of muscles of mastication in the Baikal seal (Phoca sibirica). So, in this species, the Mm, masseter, temporalis, pterygoidei, digastricus were observed in the macroscopic level. The skull characters related to these muscles were also compared between the Baikal seal and a close-related species, the ringed seal (Phoca hispida). The Mm, masseter and temporalis were well-developed using the enlarged attachment area of zygomatic arch. In contrast, the M. digastricus is suggested to be not so important in the Baikal seal, because the temporal bone is not so developed as in the ringed seal. It is suggested that the Baikal seal has especially developed the Mm. temporalis and masseter using an enlarged area of zygomatic arch among Pusa species. We also suggest that the robust temporal bone is equipped to have the M. digastricus developed as a main retractor of mandibular body in the ringed seal.

Adaptation, Biological↗

Nasal mites (Acari:Halarachnidae) in the spotted seal, Phoca largha Pallas, and other pinnipeds of Alaskan waters.

The nasal passages of 349 pinnipeds of seven species were examined for halarachnid mites. Each of seven Steller sea lions (Eumetopias jubatus), 75 of 99 harbor seals (Phoca vitulina), and four of 71 spotted seals (P. largha) were infected, but none of 28 walruses (Odobenus rosmarus), 43 ringed seals (Phoca hispida), 58 ribbon seals (P. fasciata), and 43 bearded seals (Erignathus barbatus) was infected. The sea lions harbored Orthohalarachne attenuata (Banks, 1910) and O. diminuata (Doetschman, 1944), and the harbor and spotted seals harbored Halarachne halichoeri Allman, 1847. The finding of H. halichoeri in the spotted seals is a new host record.

Alaska↗

[Neutralizing morbillivirus antibodies in the population of the Baikal seal Phoca sibirica].

The antibodies neutralizing Baikal seal morbillivirus (BSMV) were studied in sera from 148 normal Baikal seals (Phoca sibirica). The virus-neutralizing antibodies were demonstrated in sera of 61.5% of the animals examined. A comparative analysis of antibodies in NT and cell-ELISA method is presented. The role of BSMV in morbidity and mortality of seals (Phoca sibirica) is discussed.

Animals↗

Analysis of organochlorines in harbor seal (Phoca vitulina) tissue samples from Alaska using gas chromatography/ion trap mass spectrometry by an isotopic dilution technique.

A gas chromatography/ion trap mass spectrometry (GC/ITMS) method was developed for the determination of organochlorine pesticides (OCPs), polychlorinated biphenyls (PCBs) and polychlorinated naphthalenes (PCNs) in harbor seal (Phoca vitulina) tissues. Tissue samples were homogenized, lyophilized and fortified with (13)C-PCBs 28, 123, 169 and 170, and then extracted with an accelerated solvent extractor with a mixture of hexane and methylene chloride (1:1, v/v). After lipid removal using a 40% H(2)SO(4)-modified silica gel column, all organochlorines were collected in one fraction and further fractionated with an activated carbon/silica gel (1:20) column into a first fraction containing OCPs, non-coplanar PCBs and (13)C-PCBs 28, 123 and 170, and a second containing PCNs, coplanar PCBs and (13)C-PCB 169. Prior to GC/MS/MS analysis, (13)C-PCB 169 was added into the first fraction as an injection standard and (13)C-PCB 170 into the second fraction to calibrate the recoveries of the fortified internal standards. This method can effectively eliminate matrix interferences, and has high selectivity and sensitivity. Recoveries averaged 45-86% for OCPs with relative standard deviations (RSDs) of 2-14%, 52-137% for PCBs with RSDs of 3-29% and 36-152% for PCNs with RSDs of 7-29% from lard and chicken heart samples, which were used as alternative matrices to harbor seal samples in recovery studies. The limits of detection for OCPs, PCBs and PCNs were 0.7-1.9, 1.5-8.9 and 0.5-10 pg/g dry weight, respectively. This method can be used to analyze OCPs, PCBs and PCNs in harbor seal blubber, liver and kidney samples.

Alaska↗

Volume density and distribution of mitochondria in harbor seal (Phoca vitulina) skeletal muscle.

Recent studies have shown that harbor seals (Phoca vitulina) have an increased skeletal muscle mitochondrial volume density that may be an adaptation for maintaining aerobic metabolism during diving. However, these studies were based on single samples taken from locomotory muscles. In this study, we took multiple samples from a transverse section of the epaxial (primary locomotory) muscles and single samples from the m. pectoralis (secondary locomotory) muscle of five wild harbor seals. Average mitochondrial volume density of the epaxial muscles was 5.6%, which was 36.6% higher than predicted for a terrestrial mammal of similar mass, and most (82.1%) of the mitochondria were interfibrillar, unlike athletic terrestrial mammals. In the epaxial muscles, the total mitochondrial volume density was significantly greater in samples collected from the deep (6.0%) compared with superficial (5.0%) regions. Volume density of mitochondria in the pectoralis muscle was similar (5.2%) to that of the epaxial muscles. Taken together, these adaptations reduce the intracellular distance between mitochondria and oxymyoglobin and increase the mitochondrial diffusion surface area. This, in combination with elevated myoglobin concentrations, potentially increases the rate of oxygen diffusion into mitochondria and prevents diffusion limitation so that aerobic metabolism can be maintained under low oxygen partial pressure that develops during diving.

Aerobiosis↗

Polychlorinated naphthalenes and coplanar polychlorinated biphenyls in tissues of harbor seals (Phoca vitulina) from the northern Gulf of Alaska.

Blubber, liver and kidney samples of harbor seals (Phoca vitulina) from the northern Gulf of Alaska were collected during 2000-2001 for the analysis of polychlorinated naphthalenes (PCNs) and coplanar polychlorinated biphenyls (CoplPCBs). On the lipid weight (lw) base, the total concentrations of PCNs (Sigma PCNs) ranged from 0.3 to 27 ng/g lw, and the total concentrations of CoplPCBs (Sigma CoplPCBs) were 3.6-546 ng/g lw in all the tissue samples. Di-ortho PCBs and mono-ortho PCBs were dominant followed by non-ortho PCBs and PCNs. Sigma Mono-ortho PCBs and Sigma di-ortho PCBs in nursing seals were apparently lower than those in male adult seals, but Sigma PCNs and Sigma non-ortho PCBs in female adults were not significantly different from those in male adults. Differences in PCNs and CoplPCBs congener profiles in female and male adult seals are apparently related to their chemical structure and properties, animal's developmental stages and physiological conditions. A large quantity of mono-ortho and di-ortho PCBs might be transferred to newborns from the nursing seals during lactation, while non-ortho PCBs and PCNs were relatively accumulative in the mother seals. Sigma PCNs and Sigma CoplPCBs in the harbor seals correlated with ages, gender, body weight and blubber thickness, but the blubber Sigma PCNs and Sigma CoplPCBs in Kodiak Island and Southern Alaska Peninsula (KIAP) did not significantly differ from those in Prince William Sound (PWS). In addition to the new PCB data, this first report on PCN contamination in Alaskan harbor seal tissues is useful for the wildlife and ecosystem management and human health protection.

Adipose Tissue↗

Organochlorines in harbor seal (Phoca vitulina) tissues from the northern Gulf of Alaska.

Polychlorinated biphenyls (PCBs), dichlorodiphenyltrichloroethane (DDT) and its metabolites, heptachlor and hexachlorocyclohexanes (HCHs) were analyzed in the harbor seal (Phoca vitulina) tissues collected from the Gulf of Alaska during 2000-2001. summation SigmaPCBs (16-728 ng/gl w) and summation SigmaDDTs (14-368 ng/gl w) were the predominant pollutants followed by summation operatorHCHs (0.56-93 ng/gl w) and heptachlor (<or=0.068-6.0 ng/gl w). Concentrations of the above organochlorines (OCs) in the liver, kidney and blubber tissues correlated with ages, sex, body weight and blubber thickness of the harbor seals. The OC concentrations were similar between the samples collected from two different regions--Prince William Sound, and Kodiak Island and Southern Alaska Peninsula. Mean levels of OCs in nursing female adults were much lower than those in male adults, which indicate that lactation transfer OCs from mother seals to newborns.

Adipose Tissue↗

PCDD/F and PCB in harbour seals (Phoca vitulina) from British Columbia: response to exposure to pulp mill effluents.

Blubber of harbour seals (Phoca vitulina) sampled from the Strait of Georgia, BC, in 1991 and 1992 contained higher concentrations of polychlorinated dibenzo-p-dioxins and furans (PCDD/F) than did samples from Quatsino Sound on western Vancouver Island. The higher proportions of 1,2,3,6,7,8-hexachlorodibenzodioxin and 2,3,7,8-tetrachlorodibenzofuran in the Strait of Georgia samples probably reflect contamination arising from the discharge of effluents from bleached kraft mills. Higher concentrations of non-ortho- and mono-ortho substituted polychlorinated biphenyls were also found in the Strait of Georgia samples, probably as a result of general industrial activity in the region. Principal components analysis showed that residues were distributed differently between males and females. Within males, concentrations of PCDD, PCDF and PCBs appeared to increase with age. Toxic equivalents (TEQ) to 2,3,7,8-tetrachlorodibenzodioxin in the Quatsino Sd. samples were close to the working guidelines for edible fish used by Health Canada; TEQ calculated for the Strait of Georgia samples exceeded these guidelines by up to a factor of five.

Adipose Tissue↗

Assessment of cytochrome P450 1A in harbour seals (Phoca vitulina) using a minimally-invasive biopsy approach.

Biomarkers of organochlorine exposure, such as the induction of cytochrome P450 1A (CYP1A), can be used to assess the impact of environmental contaminants on the health of free-ranging marine mammal populations. The objective of the present study was to measure CYP1A in skin and liver biopsies obtained from live harbour seals (Phoca vitulina). Twelve harbour seal pups, aged three to five weeks, were captured from the Fraser River estuary, British Columbia, Canada, and temporarily held in captivity. Skin ( approximately 60 mg) and liver ( approximately 40 mg) biopsies, obtained while seals were under general anaesthesia, yielded sufficient tissue for the measurement of CYP1A by immunoblot analysis and ethoxyresorufin O-deethylase activity. A short-term exposure experiment, in which harbour seals (n=3) were treated orally with beta-naphthoflavone (BNF), resulted in increased hepatic and cutaneous CYP1A protein levels, consistent with observations in other mammals. This study is the first to measure CYP1A in skin and liver biopsies from live harbour seals and to report in vivo BNF-associated CYP1A induction in a marine mammal. The results demonstrate that microsamples collected using minimally-invasive techniques can provide toxicologically-relevant information form marine mammals.

Animals↗

The influence of underwater data transmission sounds on the displacement behaviour of captive harbour seals (Phoca vitulina).

To prevent grounding of ships and collisions between ships in shallow coastal waters, an underwater data collection and communication network (ACME) using underwater sounds to encode and transmit data is currently under development. Marine mammals might be affected by ACME sounds since they may use sound of a similar frequency (around 12 kHz) for communication, orientation, and prey location. If marine mammals tend to avoid the vicinity of the acoustic transmitters, they may be kept away from ecologically important areas by ACME sounds. One marine mammal species that may be affected in the North Sea is the harbour seal (Phoca vitulina). No information is available on the effects of ACME-like sounds on harbour seals, so this study was carried out as part of an environmental impact assessment program. Nine captive harbour seals were subjected to four sound types, three of which may be used in the underwater acoustic data communication network. The effect of each sound was judged by comparing the animals' location in a pool during test periods to that during baseline periods, during which no sound was produced. Each of the four sounds could be made into a deterrent by increasing its amplitude. The seals reacted by swimming away from the sound source. The sound pressure level (SPL) at the acoustic discomfort threshold was established for each of the four sounds. The acoustic discomfort threshold is defined as the boundary between the areas that the animals generally occupied during the transmission of the sounds and the areas that they generally did not enter during transmission. The SPLs at the acoustic discomfort thresholds were similar for each of the sounds (107 dB re 1 microPa). Based on this discomfort threshold SPL, discomfort zones at sea for several source levels (130-180 dB re 1 microPa) of the sounds were calculated, using a guideline sound propagation model for shallow water. The discomfort zone is defined as the area around a sound source that harbour seals are expected to avoid. The definition of the discomfort zone is based on behavioural discomfort, and does not necessarily coincide with the physical discomfort zone. Based on these results, source levels can be selected that have an acceptable effect on harbour seals in particular areas. The discomfort zone of a communication sound depends on the sound, the source level, and the propagation characteristics of the area in which the sound system is operational. The source level of the communication system should be adapted to each area (taking into account the width of a sea arm, the local sound propagation, and the importance of an area to the affected species). The discomfort zone should not coincide with ecologically important areas (for instance resting, breeding, suckling, and feeding areas), or routes between these areas.

Acoustics↗

Deterring effects of 8-45 kHz tone pulses on harbour seals (Phoca vitulina) in a large pool.

The marine aquaculture industry suffers losses due to pinniped attacks which damage net enclosures and fish stocks. Acoustic harassment devices (AHDs) emit loud sounds which are intended to deter pinnipeds from approaching aquaculture enclosures. At present, many AHDs emit sounds in the 8-20 kHz frequency range. It is not known whether sounds of higher frequencies have a deterrent effect on seals. Therefore five captive harbour seals (Phoca vitulina) were subjected to four series of tone pulses together spanning a broad frequency range (8, 16, 32 and 45 kHz). Pulse duration was 250 ms and pulse interval was 5s. Each of the four sounds was made deterrent by increasing the amplitude. The seals reacted by swimming away from the sounds. The displacement effect of each sound was judged by comparing the animals' surface positions, and number of surfacings, during ten 45 min baseline periods with ten 45 min test periods per frequency (one frequency per day in rotation, 40 sessions in total). The seals were displaced by all four frequencies throughout the 40 trial days. The seals came to the surface more often when the test tones were produced than in the baseline periods. The initial displacement distances did not change over the 40 test days. This suggests that operating AHDs for only short periods will be more effective and less likely to result in habituation by the seals than operating them continuously. The discomfort threshold sound pressure level (SPL) was established for each of the four pulse frequencies. The acoustic discomfort threshold SPL is defined as the boundary SPL between the area that the animals generally occupied during the transmission of the sounds and the area that they generally did not enter during sound transmission. The discomfort threshold SPL may depend on the context.

Acoustic Stimulation↗

Polychlorinated biphenyls and chlorinated pesticides in harbor seals (Phoca vitulina concolor) from the northwestern Atlantic coast.

Concentrations of polychlorinated biphenyls (PCBs), dichlorodiphenyltrichloroethane (DDT) and its metabolites, chlordane-related compounds (CHLs), mirex, hexachlorocyclohexane isomers (HCHs), dieldrin, and hexachlorobenzene (HCB) were determined in blubber of harbor seals (Phoca vitulina concolor) from the northwestern Atlantic coast. PCBs, DDTs, and CHLs were the major persistent organochlorines in harbor seal blubber, while mirex, HCHs, dieldrin, and HCB were minor contaminants. Highest concentrations were found in the adult males, followed by the pups, yearlings, adult females, and fetuses. DDT and PCB concentrations have declined from the high levels reported in the early 1970s, but no declines were observed in our samples over the ten-year period 1991-2001. DDT/PCB ratios were indicative of a more rapid decline of DDTs than PCBs, while ratios of p,p'-DDE/DDT were indicative of a metabolic "weathering" of DDT. The population appears to be susceptible to disease outbreaks, as evidenced by a recurrence of viral epizootics since the late 1970s. The PCB burdens in these seals are similar to levels reported in seals from polluted regions of Europe and Asia, and exceed the estimated threshold levels for adverse reproductive and immune system effects in the species.

Adipose Tissue↗

Development and application of bioaccumulation models to assess persistent organic pollutant temporal trends in arctic ringed seal (Phoca hispida) populations.

Individual- and population-based models were developed to simulate the bioaccumulation of persistent organic pollutants (POPs) over the lifetime of ringed seals (Phoca hispida) and account for the effects of age, growth, body condition and sex (including gestation, birth and lactation). Lactational transfer was described using a milk to blubber partition coefficient, which was shown to be a function of Kow (KMB=1/(1.676+1.293.10(-7).Kow) r2=0.60, n=32, p<0.001). Simulations showed that nursing mothers could transfer between 5% and 40% of their burden of low and high KMB contaminants, respectively, to their milk over 40 days of lactation. The models were calibrated using a POPs dataset for 50 seals (29 males, 21 females) from Arviat, Nunavut. They were initially calibrated for a group of eight highly recalcitrant PCB congeners (e.g. PCBs 153, 180) assuming a baseline elimination rate (ke) of about 0.03 year-1. For congeners more prone to biotransformation a satisfactory fit could only be reached by iteratively increasing ke to values ranging from 0.05 to 2.5 year-1 for PCBs 187 and 18, respectively; thus the models became a means for estimating ke. Estimated ke values were correlated with a calculated index of susceptibility to biotransformation (ke=0.0284/Rrelr2=0.98, n=20, p<0.001). This relationship was used to estimate ke values used in simulations to hind-cast temporal trends of SigmaPCB, SigmaDDT, SigmaCHL, alphaHCH, betaHCH in ringed seal populations and to predict potential future trends. Results of these simulations showed good agreement with measured concentrations in seals from two locations in the Canadian Arctic (Holman Island, NWT and Arctic Bay, Nunavut) collected between 1972 and 2001. The models show that the combined effects of rapid juvenile growth, a fast rate of population turnover and substantial capacity to eliminate many POPs result in fairly rapid responses to changes in contaminant loading, thus making ringed seals an effective choice for monitoring spatial and temporal trends of POPs in the Arctic marine ecosystem.

Adipose Tissue↗

Trace element concentrations in the Pacific harbor seal (Phoca vitulina richardii) in central and northern California.

To determine concentrations of trace elements (THg, MeHg, Se, and Pb) in tissues of the Pacific harbor seal (Phoca vitulina richardii), live (n=186) and dead seals (n=53) were sampled throughout central and northern California from March 2003 to January 2005. There were significant differences in THg concentrations in blood and hair based on age (p<0.001). Adult male harbor seals had greater THg concentrations in their hair than adult female harbor seals (p<0.003). THg concentrations in liver increased linearly with age and delta15N (p<0.001); whereas, MeHg concentrations in liver increased exponentially until approximately 5 years of age with an asymptote at 1.3 microg/g wet weight. MeHg expressed as a percentage of THg (%MeHg) was best described by a decay function (r2=0.796, p<0.001), decreasing to a minimum at 4 years of age. Hepatic Se increased with age and was in equimolar ratios with THg in adults; whereas, molar ratio of Se:THg in pups deviated from a 1:1 ratio. Significant differences among study locations in THg concentrations in blood and hair were not detected. Assessing the possible effect of sampling location on Hg concentrations, however, was confounded and limited by lack of equal sample sizes for basic age and sex cohorts, a common dilemma in pinniped research.

Analysis of Variance↗

Reversible reproductive control in harbour seals (Phoca vitulina) with a gonadotropin-releasing hormone agonist.

Reproductive control in captive pinnipeds is an important management subject for many facilities. To date reproductive control in harbour seals (Phoca vitulina) has been achieved using anti-androgens, progestagen preparations, castration, and physical separation of the sexes. The harbour seal group at the seal station in Friedrichskoog, Germany consists of three mature females (all >10 years), one older mature male (13 years of age in 2000) and one male who reached maturity during the study (3 years of age in 2000). In 2000 the older mature male received for the first time a 3-month depot injection of a gonadotropin-releasing hormone agonist (buserelin acetetate, 9.9 mg) by subcutaneous injection. This male was subsequently given the gonadotropin-releasing hormone agonist in 2001, 2004 and 2005. The younger male reached maturity during the investigation and received burserelin for the first time in 2004 and again in 2005. No pups were born in 2001, 2002, 2005 or 2006. No reproductive control was performed in 2002 and 2003, resulting in three newborns in 2003 and 2004. Serum levels of testosterone were measured by a routine liquid chromatography coupled with mass spectometry. Pre-burserelin testosterone levels varied between 0.02 and 2.18 ng/ml. Post-burserelin levels were under the detection limit except for the first year of the investigation. No behavioural changes such as changes in social ranking and no clinical side effects were observed. This study shows that the gonadotropin-releasing hormone agonist, burserelin acetate, can be used for reversible reproductive control in harbour seals without observed side effects or detrimental behavioural changes.

Animals↗

The transmission of phocine herpesvirus-1 in rehabilitating and free-ranging Pacific harbor seals (Phoca vitulina) in California.

Phocine herpesvirus-1 (PhHV-1) causes regular outbreaks of disease in neonatal harbor seals (Phoca vitulina) at rehabilitation centers in Europe and in the U.S. To investigate transmission of this virus samples were collected from harbor seal pups during exposure studies at a Californian rehabilitation center from 1999 to 2002 and from free-ranging harbor seals off central California during the same period. The exposure studies provided evidence that PhHV-1 can be transmitted horizontally between animals most likely through direct contact with oro-nasal secretions. However vertical transmission may also occur, as adult female harbor seals were found to be shedding the virus in vaginal and nasal secretions, and premature newborn pups had evidence of early infection. Results also indicated that PhHV-1 infections were common in both free-ranging (40%, 49/121) and rehabilitating (54%, 46/85) young harbor seals, during the spring and early summer. This timing, which correlated with pupping and weaning, suggested that the majority of animals were infected and infective with PhHV-1 between pupping and breeding.

Age Factors↗