Population genetic structure of the spotted seal Phoca largha along the Coast of Hokkaido,based on mitochondrial DNA sequences

Population genetic structure of the spotted seal, Phoca largha, along coastal regions of Hokkaido was investigated, using mitochondrial DNA (mtDNA) sequences. A 571-bp fragment of the mtDNA control region and adjacent threonine and proline transfer RNA genes was sequenced from 66 seals. We categorized all individuals into three groups considering sampling area and season: twenty-four seals from the Sea of Okhotsk in winter, 11 seals from the Sea of Okhotsk coast in fall, and 31 seals from the Sea of Japan coast in winter. From the 66 animals, 57 haplotypes were identified. Compared with the harbor seal sequence, all spotted seals examined shared two deletions in the control region, which distinguished between the two species. Forty-nine haplotypes were represented by a single individual, and haplotypes shared by more than two animals were generally restricted to same sampling-groups. Phylogenetic trees did not indicate clear geographic differences among the three groups. An Analysis of molecular variances (AMOVA) did not showed any significant population genetic structure in Hokkaido spotted seals (Phi(st )= -0.003). Our results showed a high level of diversity but no genetic structure, and did not deny the possibility that seals in the Okhotsk breeding concentration mainly stayed in the fall Okhotsk and also inhabited in the winter Sea of Japan.     

Molecular analysis of parapoxvirus from a spotted seal Phoca largha in Japan

A spotted seal Phoca largha with nodular and scab lesions on the whole body was brought to an aquarium in Nagoya, Japan. We extracted DNA from the lesions and used the polymerase chain reaction (PCR) method for detecting orthopoxvirus and parapoxvirus DNA. Parapoxvirus but not orthopoxvirus DNA was detected. The partial nucleotide sequence of the envelope gene was determined from the PCR product, and the sequence was seen to be closely related to 2 parapoxvirus strains from spotted seals in Alaska, showing 100% identity at the amino acid level, with one nucleotide substitution. Virus-neutralizing (VN) antibody against canine distemper virus (CDV) was not detected in the serum, indicating that this individual was not infected with CDV or phocine distemper virus (PDV), which both have a high mortality rate for marine mammals. These results suggest that the lesions were caused by infection with pinniped parapoxvirus, and that the viruses spread and are maintained within the habitat range or populations of spotted seals from the Bering Sea to the Japan Sea. This is the first report of molecular analysis of parapoxvirus in marine mammals in Japan.     

Ice seals as sentinels for algal toxin presence in the Pacific Arctic and subarctic marine ecosystems

Domoic acid (DA) and saxitoxin (STX)-producing algae are present in Alaskan seas, presenting exposure risks to marine mammals that may be increasing due to climate change. To investigate potential increases in exposure risks to four pagophilic ice seal species (Erignathus barbatus, bearded seals; Pusa hispida, ringed seals; Phoca largha, spotted seals; and Histriophoca fasciata, ribbon seals), this study analyzed samples from 998 seals harvested for subsistence purposes in western and northern Alaska during 2005–2019 for DA and STX. Both toxins were detected in bearded, ringed, and spotted seals, though no clinical signs of acute neurotoxicity were reported in harvested seals. Bearded seals had the highest prevalence of each toxin, followed by ringed seals. Bearded seal stomach content samples from the Bering Sea showed a significant increase in DA prevalence with time (logistic regression, p = .004). These findings are consistent with predicted northward expansion of DA-producing algae. A comparison of paired samples taken from the stomachs and colons of 15 seals found that colon content consistently had higher concentrations of both toxins. Collectively, these results suggest that ice seals, particularly bearded seals (benthic foraging specialists), are suitable sentinels for monitoring HAB prevalence in the Pacific Arctic and subarctic.     

Distribution and abundance of spotted seals Phoca largha and ribbon seals Phoca fasciata in the southern Sea of Okhotsk

The distribution and abundance of spotted seals (Phoca largha) and ribbon seals (Phoca fasciata) were assessed in March and April, 2000, by aerial line-transect surveys along the southern edge of the pack ice off the coast of Hokkaido (southern Sea of Okhotsk), Japan. Five hundred and seventeen spotted seals and 107 ribbon seals were found on the total 2944 km survey line. Total abundance was estimated to be 13 653 spotted (95% CI = 6167–30 252) and 2260 ribbon seals (95% CI = 783–6607) in March, and 6545 spotted (95% CI = 3284–815 644) and 3134 ribbon seals (95% CI = 1247–17 802 512) in April. The pack ice area off Hokkaido had higher densities (0.54 seals km^–2 and 0.58 seals km^–2 in March and April, respectively) of spotted seals than those reported in eastern Sakhalin, whereas densities (0.09 seals km^–2 in March and 0.28 seals km^–2 in April) of ribbon seals were lower than those in eastern Sakhalin. The large number of spotted seal pups suggests that the study area is an important breeding center. A greater number of female spotted seals with pups tended to be found in the center of larger and rougher floes than in other categories, and they were more abundant in stable pack ice areas. Observations of ribbon seals were limited because the survey period preceded the peak of pupping season. Ribbon seal surveys were also hampered by the inability to fly over the main breeding area between the Shiretoko Peninsula and Kunashiri Island.     

基于卫星信标跟踪的斑海豹放流效果研究

利用卫星标记跟踪方法对斑海豹的野外释放效果进行了研究。2010 年和2011 年6 月分别释放了4 头和3头人工繁殖的2 龄未成年斑海豹,2011 年同时释放了3 头野外出生的救助个体。标记斑海豹在释放后,7 头人工繁殖斑海豹中的5 头信标信号持续时间较长,在信号消失前,1 头斑海豹一直在渤海海域活动,另4 头沿辽宁沿岸、朝鲜西海岸到达辽东湾斑海豹的主要度夏海域韩国白翎岛附近。研究期间,1 头人工繁殖的斑海豹在59 d内运动的距离超过1 250 km。救助斑海豹中,2 头个体的信标信号持续较长,并分别在山东半岛沿海和黄渤海活动。研究结果表明,人工繁殖的斑海豹在经过野化训练后,放归自然海域后可以正常生活洄游。     

Movements and behavior of satellite-tagged spotted seals (Phoca largha) in the Bering and Chukchi Seas

Satellite-linked tags were attached to 12 spotted seals (Phoca largha) captured at a coastal lagoon in the eastern Chukchi Sea during August 1991–1993. Movements of seals were tracked for 32–298 days using the Argos system. Of 9,651 total location records obtained, 7,268 were usable. Individual seals were located on 41–96% of the days that tags were operational. During August–November, tagged seals alternated haul-outs at coastal sites lasting 1–304 h with trips to sea of 14–901 h. Coastal haul-outs occurred at 14 sites in western Alaska and eastern Russia. On several trips to sea, seals covered distances of more than 1,000 km. Movement southward from the Chukchi Sea generally began in October, with most of the seals passing through the Bering Strait during November. Seals first hauled out on sea ice in October (Chukchi Sea) or November (Bering Sea), and generally moved southward during October–December as sea-ice coverage increased. Seven seals, whose transmitters were still operating, spent December to June in the Bering Sea region between Kuskokwim Bay and Anadyr Gulf, which corresponded to the location of the ice front. The seals made active east-west movements within the ice front. Spotted seals are unlike other ice-breeding seals in that they regularly use coastal haul-outs during summer and autumn. Compared to the closely related Pacific harbor seal (Phoca vitulina richardsi), spotted seals make much longer trips to sea and spend longer continuous periods at their haul-outs during summer and autumn. Received: 9 April 1997 / Accepted: 30 September 1997     

Quantitative assessment of species identification in aerial transect surveys for ice‐associated seals

Technological advances have facilitated collection of vast quantities of photographic data from aerial surveys of marine mammals. However, when it is difficult to distinguish species from a distance, reliable identification from aerial images can often be challenging. This is the case for ice‐associated seals, species for which global climate change has motivated intensive monitoring efforts in recent years. We assess species and age class identification from aerial images of four ice seal species (bearded seals, Erignathus barbatus; ribbon seals, Histriophoca fasciata; ringed seals, Pusa hispida; spotted seals, Phoca largha) in the Bering Sea. We also investigate the specific phenomenological and behavioral traits commonly associated with species identification and observer confidence. We generally found species and age class misidentification occurred at relatively low levels, but only 83% of spotted seals tended to be correctly identified (with 11% mistaken as ribbon seals). We also found certain traits were strong predictors for observed species, age class, or observer confidence. Our findings add to the growing body of evidence that species misidentification is pervasive in passive sampling of animal populations. Even low levels of misidentification have been demonstrated to induce substantial biases in estimators of species distribution and abundance, and it is important that statistical models account for such errors.     

Trichinella nativa in grey seal Halichoerus grypus: spill-over from a highly endemic terrestrial ecosystem

Trichinella spp. infections of marine mammals pose a human health risk in Arctic regions where game meat is an important part of the diet. In the last decade, the grey seal population of the sub-arctic Baltic Sea has reached a level at which hunting is possible and seal meat is now available for food. We conducted a survey on the occurrence of Trichinella spp. in grey seals Halichoerus grypus (n = 171) and ringed seals Phoca hispida botnica (n = 56) of the Baltic Sea in the coastal waters of Finland, a highly Trichinella sp. endemic area. Muscle samples were examined by a mechanically assisted digestion method during 2006-2010. One grey seal was positive for Trichinella nativa , while all samples from ringed seals were negative. Even though just 1 grey seal was infected, the finding here emphasizes the importance of proper meat inspection of seals intended for human consumption, especially in areas with high infection pressure.     

Feeding ecology of phocid seals and some walrus in the Alaskan and Canadian Arctic as determined by stomach contents and stable isotope analysis

Feeding habits of ringed (Phoca hispida), bearded (Erignathus barbatus), spotted (Phoca largha) and ribbon (Phoca fasciata) seals and walrus (Odobenus rosmarus) were studied using stomach contents and stable carbon and nitrogen isotopes. Bearded seals fed benthically, primarily crustaceans and mollusks. Both zooplankton and fish were significant prey for ringed seals, while fish was principal spotted seal prey. Few gastric contents were available from ribbon seals. δ¹⁵N was positively correlated with age in ribbon seals and δ¹³C was positively correlated with age in ringed and ribbon seals. δ¹⁵N was highest in spotted seals, in agreement with their fish-dominated diet. δ¹⁵N was not different between Alaskan-harvested ringed and bearded seals, while δ¹⁵N was lowest in ribbon seals and walrus. Carbon-13 was most enriched in bearded seals and walrus reflecting benthic ecosystem use. Canadian ringed seals were depleted in ¹³C compared to Alaskan pinnipeds, likely because of Beaufort Sea versus Chukchi and Bering seas influence.     

辽东湾斑海豹MHC-DQB 基因的遗传多样性分析

本文从25 份斑海豹样本中获得141 bp 片段,发现21 个变异位点,定义了12 个MHC-DQB 等位基因,氨基酸变异率为25.5% 。等位基因之间的遗传距离范围是0. 0071 ～ 0.1064,平均值为0.0577,不同等位基因之间的碱基差异是1 ～ 15 bp,平均差异数为8 bp。与其他鳍足类动物对比后发现,斑海豹MHC-DQB 表现出较丰富的多态性。非同义替换率明显高于同义替换率,由此造成的氨基酸替换集中在肽结合位点PBR 附近,表明DQB 基因受到强烈的平衡选择作用。11 个样本出现多于两条等位基因的情况,推测存在基因重复现象。     

Bioaccumulation of lithium by marine organisms in European, American, and Asian coastal zones: microanalytic study using secondary ion emission

37 species of aquatic mammals, fish, crustacea, annelids, molluscs belonging to cephalopods, gasteropods and lamellibranchs were collected from coastal waters of France: North Sea, English Channel, Atlantic Ocean, Mediterranean Sea, from Greece: Aegian Sea, from North America: Atlantic Ocean and from Japan: Pacific Ocean, Sea of Japan and East China Sea. Microanalyses which were performed on organs and tissues, using secondary ion mass spectrometry, revealed high concentrations of lithium, which is commonly used in human therapy, but is also toxic in low amounts. The retention of this metal by the marine organisms appears as a general phenomenon independent of their biotope and geographical origin; the highest lithium levels were detected in the fish muscles (= edible part).     

Resource partitioning by sympatric pagophilic seals in Alaska: monitoring effects of climate variation with fatty acids

Four species of pagophilic phocid seals in the Western Arctic—bearded (Erignathus barbatus), ringed (Phoca hispida), ribbon (Phoca fasciata), and spotted (Phoca largha)—are particularly vulnerable to arctic warming trends. Documenting diet composition over time is one way in which the flexibility of a species, in the face of broad ecosystem changes, can be measured. Because the fatty acid (FA) composition of depot lipids has long been known to reflect diet, we analysed the FA composition of blubber samples collected from bearded (n = 30), ringed (n = 15), ribbon (n = 32) and spotted seals (n = 24). All animals were taken near Little Diomede Island in May and June 2003, providing a unique opportunity to study interspecies differences with minimal effects of spatial and temporal variation. Bearded seal FA composition was significantly different from all other seals (P < 0.001 in all cases), ringed seal FA composition was significantly different from that of both ribbon and spotted seals (P < 0.001), but ribbon and spotted seal FA compositions could not be distinguished from each other. Overall, the blubber FA compositions of ribbon, ringed and spotted seals implied diets dominated by pelagic prey. Inferences we made about current diets of bearded and ringed seals suggest that certain prey important to them in this area in the 1970s remain important today, despite notable changes in the ecosystem. We believe that blubber FA analysis will be a useful tool in assessing the response of ice-associated pinnipeds and their ecosystem to changes associated with arctic warming.     

Surreptitious sympatry: Exploring the ecological and genetic separation of two sibling species

Climate change is having profound impacts on animal populations, and shifts in geographic range are predicted in response. Shifts that result in range overlap between previously allopatric congeneric species may have consequences for biodiversity through interspecific competition, hybridization, and genetic introgression. Harbor seals (Phoca vitulina) and spotted seals (Phoca largha) are parapatric sibling species and areas of co‐occurrence at the edges of their range, such as Bristol Bay, Alaska, offer a unique opportunity to explore ecological separation and discuss potential consequences of increased range overlap resulting from retreating sea ice. Using telemetry and genetic data from 14 harbor seals and six spotted seals, we explored the ecological and genetic separation of the two species by comparing their utilization distributions, distance from haul‐out, dive behavior (e.g., depth, duration, focus), and evidence of hybridization. Firstly, we show that harbor and spotted seals, which cannot be visually distinguished definitively in all cases, haul‐out together side by side in Bristol Bay from late summer to early winter. Secondly, we observed subtle rather than pronounced differences in ranging patterns and dive behavior during this period. Thirdly, most spotted seals in this study remained close to shore in contrast to what is known of the species in more northern areas, and lastly, we did not find any evidence of hybridization. The lack of distinct ecological separation in this area of sympatry suggests that interspecific competition could play an important role in the persistence of these species, particularly if range overlap will increase as a result of climate‐induced range shifts and loss of spotted seal pagophilic breeding habitat. Our results also highlight the added complexities in monitoring these species in areas of suspected overlap, as they cannot easily be distinguished without genetic analysis. Predicted climate‐induced environmental change will likely influence the spatial and temporal extent of overlap in these two sibling species. Ultimately, this may alter the balance between current isolating mechanisms with consequences for species integrity and fitness.     

Cycle of transformation of the spotted seal (<Emphasis Type="Italic">Phoca largha</Emphasis>, Pallas, 1811) onshore associations in Peter the Great Bay of the Sea of Japan

In Peter the Great Bay, the spotted Seal (Phoca largha) is closely connected with island haulout sites sites all year round and forms four types of onshore associations, preliminary, reproductive, molting, and rehabilitative. It has been found over long term studies that changes in these associations make up an annual cycle. The specific redistribution pattern of seals in the bay appears to be a result of consecutive transformation processes, viz., the break up of previous types of associations and the simultaneous formation of the following ones. The mechanisms underlying every type of onshore associations of spotted seals and the peculiarities of their functioning are discussed.     

The seasonal dynamics of diarrhetic shellfish poisoning toxins in the digestive gland of <Emphasis Type="Italic">Crenomytilus grayanus</Emphasis> (Dunker, 1853) analyzed by high-performance liquid chromatography with fluorometric detection

The seasonal dynamics of diarrhetic shellfish poisoning (DSP) toxins in the digestive gland of the mussel Crenomytilus grayanus (Dunker, 1853) from Peter the Great Bay in the Sea of Japan have been studied. Toxins in the form of 4-bromomethyl-7-methoxycoumarin derivatives have been analyzed using high-performance liquid chromatography (HPLC) with fluorometric detection. This method allows the determination of the quantitative and qualitative composition of individual toxins. All the C. grayanus samples contained DSP toxins, but their concentration varied substantially during the year. The highest concentration was recorded from the mussels collected in May; the lowest concentration was recorded in February. The toxins were represented predominantly by dinophysistoxin-2; in July-September and December, okadaic acid was detected. An assumption has been made about the relationship between the potential producers of DSP toxins in the waters of the Peter the Great Bay and the dynamics of these toxins in tissues of C. grayanus. The HPLC method, as an alternative to the enzyme-linked immunosorbent assay, was used to assess the quantitative and qualitative composition of DSP toxins in seafood for the first time in the Russian Federation. This method can be recommended for the routine monitoring of toxins conducted by the controlling organizations of the Russian Federal Service for Veterinary and Phytosanitary Surveillance.     

Distribution and selective elimination of paralytic shellfish toxins in different tissues of Octopus vulgaris

Octopus (Octopus vulgaris, Cuvier) plays a central role in the marine food web, being an important consumer with high metabolic rates and at the same time an important food item for higher predators. After harmful algal blooms, octopus can accumulate high levels of marine toxins trough trophic interrelationships. The aim of this study is to characterize the distribution of paralytic shellfish toxins (PSTs) in selected tissues of the O. vulgaris, in order to assess the translocation of toxins among organs with different physiological functions. Different retention times and selective elimination of particular toxin analogues were also investigated. Twenty three specimens of O. vulgaris were captured in Peniche (NW coast of Portugal) after PSTs have been detected in molluscan bivalves. Tissue matrices were dissected from organs with digestive function (digestive gland, stomach and salivary glands) and excretory function (kidneys and branchial hearts) and analyzed for toxin determination. Toxin analysis was carried out by high performance liquid chromatography with fluorescence detection (LC-FLD). PSTs were found in all tissues analyzed. The highest toxin concentrations were found in the digestive gland, reaching a maximum of 2980 μg STX equiv. kg⁻¹. The toxin profile was constituted by dcSTX, B1, C1 + 2, dcGTX2 + 3, dcNEO, STX and GTX2 + 3. A lower number of toxins were identified in the remaining organs, with B1 and dcSTX compromising more than 90% in molar fraction. Decarbamoyl saxitoxin was the most abundant toxin detected in digestive gland, stomach and salivary glands, while B1 was dominant in organs with excretory function. A positive correlation of concentrations of B1 and dcSTX were found in the organs analyzed. Results indicate that B1 and dcSTX are assimilated into the digestive gland in a similar proportion. Selective elimination of toxins with higher elimination of B1 and retention of dcSTX is suggested. This study contributes to better understanding of the dynamics of PSTs in O. vulgaris and the fate of PSTs in the food web.     

Assessing the genetic effects of rehabilitating harbor seals (<Emphasis Type="Italic">Phoca vitulina</Emphasis>) in the Wadden Sea using stochastic simulations

Rehabilitation of marine mammals with the intent of releasing them back into nature is carried out for several species. Rehabilitation can help supporting critically endangered species, increase public awareness, and serve scientific purposes, but rehabilitation also causes concern due to risk of introducing diseases into the wild populations and genetic changes from disruption of natural selection and host-pathogen co-evolution. In this study, we investigate another potential risk from rehabilitation that has not previously been considered, i.e., anthropogenic increase in population inbreeding level. For this purpose, we performed stochastic population viability analyses of the Wadden Sea harbor seal population using VORTEX. In the Wadden Sea, rehabilitation takes place in several rehabilitation centers, and many of the seals suffer from parasitic bronchopneumonia or are orphaned. Several studies have found a correlation between helminth infection and inbreeding level. Moreover, a relation between fitness traits such as pup survival and inbreeding has been demonstrated. On this basis, we assess the effects from rehabilitating relatively inbred seals on population size and genetic diversity. We find that releasing seals significantly affect population inbreeding level and to a lesser extent population size. The effect depends on the level of inbreeding in rehabilitated seals as well as the actual number of inbred seals released.     

Kunitz-type protease inhibitors from acrorhagi of three species of sea anemones

Sea anemones are rich in biologically active polypeptides such as toxins and protease inhibitors. These polypeptides have so far been isolated from whole bodies, tentacles or secreted mucus. Recently, two novel peptide toxins with crab lethality have been isolated from acrorhagi (specialized aggressive organs elaborated by only certain species of sea anemones belonging to the family Actiniidae) of Actinia equina. This prompted us to survey biologically active polypeptides in the acrorhagi of two species of sea anemones, Anthopleura aff. xanthogrammica and Anthopleura fuscoviridis. No potent crab lethality was displayed by the acrorhagial extracts of both species. However, significantly high protease inhibitory activity was instead detected in the acrorhagial extracts of the two species and also in that of A. equina. From the acrorhagi of A. equina, A. aff. xanthogrammica and A. fuscoviridis, one (AEAPI), one (AXAPI) and two (AFAPI-I and AFAPI-III) protease inhibitors were isolated, respectively. The complete amino acid sequences of the four inhibitors were elucidated by N-terminal sequencing and sequencing of the C-terminal peptide fragment produced upon asparaginylendopeptidase digestion. The determined amino acid sequences revealed that all the four inhibitors are new members of the Kunitz-type protease inhibitor family.     

Heavy Metals in the Tissues of Commercially Important Fish of Amurskii Bay,Sea of Japan

The concentrations of iron, copper, zinc, manganese, cadmium, and mercury in tissues and organs of Pacific herring, Far Eastern navaga, and spotted flounder from Amurskii Bay, Sea of Japan, were determined using an atom-absorption method. The distribution of these elements has been studied in organisms of the fish. The greatest concentration of iron, copper, cadmium, and mercury is found in the liver of the fish, manganese is mostly accumulated in the bone tissue, and zinc is found in the skin. Some specific features of metal accumulation in the fish of Amurskii Bay have been revealed. For example, the concentration of iron in the liver of herring and flounder significantly increased the mean concentration known from other areas. A sanitary–hygienic evaluation is provided for the recent levels of metal concentrations in these three species of commercially important fish.     

Serologic evidence of Brucella infection in pinnipeds along the coast of Hokkaido,the northernmost main island of Japan

Brucella infection in Hokkaido was serologically surveyed in four species of pinnipeds inhabiting Cape Erimo during 2008–2013 and the Shiretoko Peninsula in 1999 by ELISA using Brucella abortus and B. canis as antigens. Anti‐Brucella positive sera showed higher absorbance to B. abortus than B. canis in almost all samples. Anti‐B. abortus antibodies were detected in serum samples from 24% (n = 55) of Western Pacific harbor seals (Phoca vitulina stejnegeri) in Cape Erimo and from 66% (n = 41) of spotted seals (P. largha), 15% (n = 20) of ribbon seals (Histriophoca fasciata) and 18% (n = 17) of Western Steller's sea lions (Eumetopias jubatus jubatus) in the Shiretoko Peninsula. Anti‐Brucella antibodies were detected at higher absorbance in 1‐ to 4‐year‐old harbor seals than in the pups and mature animals, suggesting either that Brucella infection mainly occurs after weaning or that it is maternally transmitted to pups with premature or suppressed immunity. Anti‐Brucella antibodies were detected in both immature and mature spotted seals and ribbon seals, with higher absorbance in the former. The antibodies were detected only in mature Western Steller's sea lions. Western blot analysis of the serum samples showed some differences in band appearances, namely discrete versus smeary, and in the number of bands, indicating that multiple different Brucella may be prevalent in pinnipeds in Hokkaido. Alternatively, the Brucella of pinnipeds may have some intra‐species diversity.