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Contamination levels and specific accumulation of persistent organochlorines in Caspian seal (Phoca caspica) from the Caspian sea, Russia.

Persistent organochlorines, such as polychlorinated biphenyls (PCBs) including coplanar congeners, DDTs, HCHs, chlordanes (CHLs), and HCB, were determined in the blubber of Caspian seals (Phoca caspica) and their fish diet (Rutilus sp.) collected in 1993 from the northern Caspian Sea, Russia. Notable concentrations of DDTs and HCHs were found at mean values of 19 and 1.3 microg/g on wet-weight basis in adult male seals, respectively. PCB pollution in Caspian seals was not so considerable compared with those of seals that suffered mass mortality. Less gender difference of organochlorine residue levels in adult animals implies less excretion of organochlorines from the body of adult females through lactation and gestation, probably due to the higher rate of pregnancy failure. Immature seals had a wide range of organochlorine concentrations, which decreased as body length increased, suggesting dilution. Caspian seals can be considered to have higher degradation capacity for coplanar PCBs. Mean TEQs (2,3,7,8-TCDD toxic equivalents) for non-, mono- and di-ortho coplanar PCBs was 51 pg/g on wet-weight basis, which was lower than those in seals that have suffered mass mortality, but comparable to those found in Arctic seals.

Age Factors↗

Experimental evidence of seawater drinking in juvenile hooded (Cystophora cristata) and harp seals (Phoca groenlandica).

This study was undertaken to measure whether young harp seals (Phoca groenlandica) and hooded seals (Cystophora cristata) drink seawater and, if so, to investigate how the excess salt load is handled. Blood and urine samples were collected from hooded seal pups (n = 3) and harp seal pups (n = 3) after 2 weeks of freshwater exposure, at intervals during 3 weeks of seawater exposure and, finally, after 2 weeks of re-exposure to fresh water. Total water turnover, as measured by injection of tritiated water, was 2200 ml x day(-1) and 3300 ml x day(-1) in hooded seals and harp seals, respectively. The extent of mariposia was taken as the difference between total water turnover and influx of water through food (free and metabolic water) and respiratory water exchange. Seawater drinking amounted to 14% and 27% of total water turnover (rH2O) for the hooded seals and harp seals, respectively. Further evidence of mariposia was obtained from an increase in the excretion rate of the urine osmolytes Na+, Cl- and Mg2+, during the period of seawater exposure. It is concluded that water influx due to seawater drinking can not be excluded as a source of error when estimating food consumption of free-ranging harp seals and hooded seals, by use of labeled water techniques.

Age Factors↗

Melatonin potentiates the vasoconstrictive effect of noradrenaline in renal artery from newborn hooded seals (Cystophora cristata) and harp seals (Phoca groenlandica).

Isometric tension was recorded by force displacement transducers in ring segments of the inferior branch of the renal artery from newborn hooded seals (Cystophora cristata, n=6), harp seals (Phoca groenlandica, n=3) and domestic pigs (Sus scrofa f. domestica, n=5). Arterial segments were mounted in heated organ baths and exposed to graded concentrations of noradrenaline (NA) and adrenaline (A), alone or together with melatonin. The melatonin concentration in the bath was similar to the plasma concentration normally found in each experimental animal. Melatonin alone did not affect the tension in any of the species, but melatonin potentiated the contraction induced by NA in hooded and harp seal arteries to a maximum of about 25% above the resting, pre-stretch level. The selective melatonin receptor antagonist luzindole reduced this potentiation by 80%. Species-specific concentrations of melatonin did not potentiate the NA-effect in the domestic pig or the A-induced contraction in any of the species. The results indicate that melatonin specifically increases the NA-sensitivity of smooth muscles in renal arteries from newborn seals. It is presumed that similar effects may operate in foetal and maternal seals and may influence their circulation during maternal diving.

Animals↗

Chromosomal homeologies between human, harbor seal (Phoca vitulina) and the putative ancestral carnivore karyotype revealed by Zoo-FISH.

We report on the construction of the first comparative Zoo-FISH map of a marine mammal. Zoo-FISH with DNA probes from a human chromosome-specific library to metaphase spreads of the harbor seal (Phoca vitulina) disclosed 31 conserved syntenic segments covering the complete autosomal complement and the X chromosome. Comparison with Zoo-FISH maps of other species reveals that the harbor seal shares a high degree of karyotypic homeology with the human complement and an even higher degree with the conordinal cat complement. These findings suggest that pinniped, felid and human karyotypes have maintained conserved complements. Based on data of Zoo-FISH and comparative cytogenetics, a Zoo-FISH map of the ancestral carnivore karyotype (Z-CAR) is proposed. Flow cytometry revealed that the DNA value of the harbor seal genome is 79% that of the human genome.

Animals↗

Intestinal helminths of a landlocked ringed seal (Phoca hispida saimensis) population in eastern Finland.

A small, landlocked, endangered ringed seal (Phoca hispida saimensis) population lives as a postglacial relict in Lake Saimaa in eastern Finland. In this study, the intestinal metazoans were examined from a total of 61 Saimaa seals found dead from 1981 to 2001. The helminth fauna was very depauperate. Only one acanthocephalan species, Corynosoma magdaleni, has been able to survive during isolation in the freshwater environment. In addition, only two cestode species were found: Diphyllobothrium ditretum and Schistocephalus sp. However, neither of these larvae developed in the ringed seals. As the newborn pups of Saimaa seals are nursed for about 2 months only one of them was infected by C. magdaleni. At 2-5 months the pups harboured worms; however, the mean intensity did not rise distinctively until the seals were adults. C. magdaleni infection increased with increasing age and the abundances were significantly different between the age groups. There was no correlation between the number of parasites and the adult seal body weight; however, seals weighing 36 kg or more carried the majority of the worms. The distribution of the parasites was aggregated; six seals carried 65% (n=1,182) of all worms (total n=1,809) and 26 seals were uninfected. Variation in the number of worms in individual seals may indicate differences in feeding behaviour and in the diet of seals, and/or variation in the immune status of the seals.

Acanthocephala↗

Sealworm (Pseudoterranova decipiens) infection in common seals (Phoca vitulina) and potential intermediate fish hosts from the outer Oslofjord.

Infection with Pseudoterranova decipiens in 45 common seals (Phoca vitulina) from the outer Oslofjord was investigated. Seals were collected before and during the phocine distemper virus epizootic in 1988 when the seal population in the area was reduced from about 350 to 100. In 1989, cod (Gadus morhua) and other fish species were sampled for comparison with earlier investigations on prevalence and abundance of P. decipiens infection in fishes from the outer Oslofjord. Seventy-two per cent of cod were infected with P. decipiens larvae in shallow waters between the seal skerries; the corresponding abundance was 2.2. At other fishing sites at increasing distances from the seal colony, abundance dropped to 0.05-0.20. Otoliths recovered from stomachs and scats indicated that gadids (Micromesistius poutassu, Trisopterus esmarkii and G. morhua) made up more than 80% of the seal diet. Bullrout (Myxocephalus scorpius) was also heavily infected with P. decipiens (prevalence 93% and abundance 8.4), but was not found in the seal diet. However, uninfected cod that enter shallow water from the surrounding deep waters became infected when they feed on bullrout. Recruitment to mature stock of P. decipiens occurs when highly infected cod are eaten by seals. The number of reproducing P. decipiens is very low in common seals. Only 2 seals (i.e. less than 5% of the sample) were simultaneously infected with mature worms of both sexes. The dramatic reduction of the seal stock in the outer Oslofjord by the epizootic did not seem to affect the abundance of P. decipiens in its intermediate hosts.

Animals↗

Short term fasting does not aggravate immunosuppression in harbour seals (Phoca vitulina) with high body burdens of organochlorines.

Two groups of 11 harbour seals (Phoca vitulina) with different body burdens of organochlorines were subjected to an experimental 15-day fasting period, during which they lost an average 16.5% of their body weights. Blood levels of the most persistent organochlorines showed an approximate twofold increase, while levels of aryl hydrocarbon receptor-binding organochlorines remained largely unaffected. Few differences in immunological parameters were observed between the two dietary groups. Numbers of circulating lymphocytes dropped to about 65% of the initial values and NK cell activity showed a slight increase in both groups. Mitogen- and antigen-induced lymphoproliferative responses of the Baltic group of seals remained within normal ranges. These results suggest that relatively short-term fasting periods do not present an additional immunotoxicological risk to seals with high body burdens of organochlorines.

Adipose Tissue↗

Organochlorine levels in common seals (Phoca vitulina) which were victims and survivors of the 1988 phocine distemper epizootic.

We compared concentrations of organochlorines in the blubber of common seals (Phoca vitulina) found dead during the 1988 phocine distemper epizootic with levels in animals which survived it. There were highly significant differences between the live and dead animals, and between sample sites. These were not fully accounted for by seasonal and condition-related changes in blubber thickness.

Adipose Tissue↗

The relationship of the increased level of metallothionein with heavy metal levels in the tissue of the harbor seal (Phoca vitulina).

The relationship between age, heavy metal levels, and a heavy-metal binding protein, metallothionein (MT), in the liver and kidney of the harbor seal (Phoca vitulina) was studied. The cadmium (Cd) level in the liver and the Cd, inorganic mercury (I-Hg), and zinc (Zn) levels in the kidney were increased with body length, suggesting an age-related accumulation of these metals. The MT levels determined by an MT radioimmunoassay showed concentrations of 240 +/- 139 micrograms/g in the liver and 343 +/- 219 micrograms/g in the kidney. These MT levels were shown to be correlated with age. The statistically significant relationship of the MT levels with the levels of Cd and Zn in the liver and those of Cd, Zn, and I-Hg in the kidney suggested that the protein is responsible for the sequestration of these metals as already observed in terrestrial animals.

Animals↗

Mitogen and antigen induced B and T cell responses of peripheral blood mononuclear cells from the harbour seal (Phoca vitulina).

In vitro assays were developed for studies concerning the functioning of the immune system of the harbour seal (Phoca vitulina). Proliferative responses of peripheral blood mononuclear cells (PBMC) were measured after stimulation with different concentrations of the mitogens concanavalin A (Con A), pokeweed mitogen (PWM), phytohaemagglutinin (PHA) or lipopolysaccharide from Salmonella typhimurium (LPS). Con A and PWM induced strong proliferative responses, while PHA and LPS induced comparatively low proliferative responses. Responses of mitogen stimulated PBMC to recombinant human interleukin-2 (rhIL-2) and in vitro immunoglobulin production by mitogen stimulated PBMC were measured to discriminate between stimulation of T cells and B cells. It was found that Con A and PHA stimulate phocine T cells, PWM stimulates both T cells and B cells and LPS predominantly stimulates phocine B cells. Antigen-specific immune responses were measured after immunization of seals with an inactivated rabies vaccine and/or with tetanus toxoid. Antigen-specific proliferation of PBMC and the presence of antigen-specific antibody forming cells were demonstrated for both antigens in the PBMC of immunized animals. The responses measured in vitro correlated well with the development of specific serum antibody titers to these antigens.

Animals↗

Vaccination of harbour seals (Phoca vitulina) against phocid distemper with two different inactivated canine distemper virus (CDV) vaccines.

Two inactivated canine distemper virus (CDV) vaccines--an adjuvanted whole inactivated virus and a subunit ISCOM preparation--were tested for their ability to induce protective immunity in harbour seals (Phoca vitulina) against phocid distemper, a disease that recently killed greater than 17,000 harbour seals in the North and Baltic seas, and was shown to be caused by infection with a newly discovered morbillivirus, which is antigenically closely related to CDV. Four CDV seronegative harbour seals were vaccinated three times with the whole-virus vaccine, two with the ISCOM subunit vaccine and two were sham-vaccinated with an antigen-free preparation. Ten days after the last vaccination, when all six vaccinated animals had developed CDV neutralizing antibody titres ranging from 300 to 3000, all eight animals were challenged by the oculonasal and the peritoneal routes, with an organ suspension from dead seals. None of the six vaccinated animals developed clinical signs. The two sham-vaccinated seals died on days 14 and 18, respectively, after having shown a body temperature rise, respiratory symptoms and weight loss. In organs from both dead animals morbillivirus antigen was demonstrated with an enzyme-linked immunosorbent assay and an immunofluorescence assay. One of these two animals had developed a low titre of CDV-specific antibodies just before death. These data clearly indicate that seals can be protected from fatal challenge with the phocid distemper virus (PDV), by vaccination with certain inactivated CDV vaccines. They also reconfirm that infection with PDV should be considered the primary cause of the recent epizootic in seals.

Animals↗

Canine distemper virus ISCOMs induce protection in harbour seals (Phoca vitulina) against phocid distemper but still allow subsequent infection with phocid distemper virus-1.

A candidate canine distemper virus (CDV) ISCOM vaccine has been shown to be effective in protecting harbour seals (Phoca vitulina) from phocid distemper in 1988. However, of the 35 harbour seals receiving this vaccine upon admission to a seal rehabilitation and research centre (Pieterburen, The Netherlands) in 1989, six developed mild inflammatory symptoms of the respiratory tract. Phocid distemper virus-1 (PDV-1) could be isolated from three of these animals. This indicates that the vaccine affords protection from phocid distemper, but may still allow PDV-1 infection of the respiratory tract. Contacts with non-vaccinated seals should then be prevented until no more virus is excreted. It is speculated that this PDV-1 infection of the respiratory tract in CDV-ISCOM vaccinated seals is followed by a lifelong immunity.

Animals↗

Levels of polychlorinated biphenyls, organochlorine pesticides, mercury, cadmium, copper, selenium, arsenic, and zinc in the harbour seal, Phoca vitulina, in Norwegian waters.

Residue levels of the chlorinated hydrocarbons polychlorinated biphenyls (PCBs), total DDT, alpha-, beta- and gamma-hexachlorocyclohexane (HCH), hexachlorobenzene (HCB), and oxychlordane in blubber, and the elements mercury, cadmium, copper, selenium, arsenic, and zinc in liver, of 82 harbour seals, Phoca vitulina, were determined. The seals were found dead or dying in Norwegian waters during the disease outbreak caused by a morbilli virus in 1988. Of the chlorinated hydrocarbons, the highest concentrations were found of PCBs, which were 2-4 times higher than the total DDT concentrations. P,p'-DDE was the main contributor to the total DDT, and constituted about 80%. The PCB and total DDT concentrations ranged from 0.4-38 and 0.1-8.8 mg kg(-1), respectively. The mercury concentrations ranged from 0.1-89 mg kg(-1). Significantly higher mean levels of PCBs (13 mg kg(-1) and mercury (16 mg kg(-1)) were found in blubber and liver, respectively, of seals from the Southern coast of Norway, as compared to the corresponding mean levels in seals from the Oslofjord (8.8 and 4.1 mg kg(-1)), and at the Northwestern coast (5.8 and 7.9 mg kg(-1)), respectively. A significant positive correlation was found between the concentrations of selenium and mercury. When the seals were grouped according to sex and age, females of ageclass > 1 and pups of both sexes had significantly lower PCB and total DDT levels than males ageclass > 1. Significantly higher hepatic mercury levels were found in seals ageclass > 1 as compared to pups. Only low levels of the other organochlorines, cadmium and arsenic, were found. Copper and zinc were considered to be present at normal physiological levels. The present organochlorine and heavy metal concentrations gave no support to suggestions that organochlorines and heavy metal pollution may be directly involved in the observed seal deaths.

Journal Article↗

Levels of selected individual polychlorinated biphenyls in different tissues of harbour seals (phoca vitulina) from the Southern coast of Norway.

Concentrations of individual polychlorinated biphenyls (PCBs) were determined in blubber, liver, kidney and brain tissues of 10 male harbour seals (Phoca vitulina) of different ages. The animals were found dead or dying on the southern coast of Norway in 1988 during the morbilli virus epizootic. Twenty-three PCB congeners were investigated in all tissues by capillary gas chromatography with electron capture detection and comparison with standards of the respective congeners (IUPAC numbers 28, 52, 74, 99, 101, 105, 110, 114, 118, 128, 138, 141, 149, 153, 156, 157, 170, 180, 183, 187, 194, 206 and 209). In addition, three toxic coplanar congeners (IUPAC numbers 77, 126 and 169) were investigated in the blubber samples. Total PCB (sum of the determined congeners) in kidney (12-60 nmol g(-1)), liver (12-90 nmol g(-1)) and blubber (10-79 nmol g(-1)) were similar on extractable lipid basis, while total PCB in brain lipid was about 1/10 (1.6-12 nmol g(-1)). The absolute concentrations of the individual congeners in brain lipid correlated poorly with the respective concentrations in each of the other tissues, while the individual congener concentrations in the lipid fractions of kidney, liver and blubber tended to correlate with each other. The relative concentrations of the sum of the most persistent congeners (PCB-99, -128, -138, -153, -170, -180, -187 and -194) correlated in all tissues, and an increase of the relative concentrations of these congeners, most distinct in blubber, with higher carcass weight, was shown. Also, total PCB in blubber correlated with carcass weight. When comparing the relative concentration patterns of individual congeners (per cent of total) in the four tissues, six congeners differed significantly between tissues, and the relative brain concentrations showed most deviation from the other tissues. The concentration of the coplanar PCBs was 1 : 1000-1 : 1000 of the total PCB concentration. The results demonstrate a lower accumulation together with a different concentration pattern in the brain as compared to other tissues.

Journal Article↗

Bile acids from the harp seals, Phoca groenlandica.

1. The bile acids composition of the harp seal, Phoca groenlandica, collected around Newfoundland, Canada, had been examined. 2. 13C n.m.r. spectroscopy of the crude bile extract was superior to t.l.c. analysis in revealing the presence of phocaecholic acid and a taurine moiety in this mixture. 3. The relative percentage of cholic, phocaecholic and chenodeoxycholic acids was 60.5% (+/- 4.5), 21.4% (+/- 4.0) and 18.1% (+/- 5.8), respectively. 4. The variation in the percentage of bile acids did not correlate with the sex of the sampled animals, but varied significantly with the age of the seals.

Age Factors↗

Purification of alkaline phosphatase from the intestinal content of common seal (Phoca vitulina larga) by immunoaffinity chromatography.

1. A monoclonal antibody APP, 1 against harp seal alkaline phosphatase has been prepared. It was found that the antibody was cross-reacted with the intestinal alkaline phosphatase of common seal Phoca vitulina larga. 2. Purified antibody was linked to Sepharose 4B and used for immunoaffinity chromatographic purification of alkaline phosphatase from the intestinal content of common seal. A spec. act. of the purified enzyme was 7300 units per mg of protein. 3. The enzyme in 7.5% polyacrylamide gel in the presence of 2-mercaptoethanol and SDS was migrated as a single band of Mr 67,000. The value of the apparent Km for common seal alkaline phosphatase was equal to 3.7 mM.

Alkaline Phosphatase↗

Fatty acid composition of fats around the mystacial and superciliary vibrissae differs from that of blubber in the Saimaa ringed seal (Phoca hispida saimensis).

1. Adipose tissues associated with the mystacial and superciliary vibrissae of the Saimaa seal (Phoca hispida saimensis) contained 31% of hexadecenoic acid (16:1 omega 7), which was almost twice the already high amount in blubber. The amount of tetradecenoic acid (14:1 omega 5) was also double that in blubber. 2. At the same time the levels of long-chain highly unsaturated docosapentaenoic (22:5 omega 3) and docosahexaenoic acids (22:6 omega 3) were only two-thirds of those found in blubber. 3. It is suggested that the excess of low-melting monoenoic fatty acids (14:1 and 16:1) in adipose tissues near the vibrissae might not be compensation for the low supply of dietary polyunsaturated fatty acids in these tissues, but that the self-induced excess of monoenoic fatty acids ensured the proper functioning of these well-developed sense organs.

Adipose Tissue↗

Characterisation of morbilliviruses isolated from Lake Baikal seals (Phoca sibirica).

Sequence analysis of the haemagglutinin protein (H) gene of the morbillivirus (PDV-2) isolated from a Siberian seal (Phoca sibirica) during the 1987/1988 epizootic in Lake Baikal revealed that it was most closely related to two recent isolates of canine distemper virus (CDV) from Germany and different from CDV vaccines currently in use in that region. The virus continued to circulate in seals in Lake Baikal after the 1987/1988 epizootic since sera collected from culled seals in the spring of 1992 were positive in morbillivirus ELISA tests, reacting most strongly with the CDV antigen.

Amino Acid Sequence↗