Anna Roos

Mitochondrial Control Region and microsatellite analyses on harbour porpoise (Phocoena phocoena) unravel population differentiation in the Baltic Sea and adjacent waters

The population status of the harbour porpoise (Phocoena phocoena) in the Baltic area has been a continuous matter of debate. Here we present the by far most comprehensive genetic population structure assessment to date for this region, both with regard to geographic coverage and sample size: 497 porpoise samples from North Sea, Skagerrak, Kattegat, Belt Sea, and Inner Baltic Sea were sequenced at the mitochondrial Control Region and 305 of these specimens were typed at 15 polymorphic microsatellite loci. Samples were stratified according to sample type (stranding vs. by-caught), sex, and season (breeding vs. non-breeding season). Our data provide ample evidence for a population split between the Skagerrak and the Belt Sea, with a transition zone in the Kattegat area. Among other measures, this was particularly visible in significant frequency shifts of the most abundant mitochondrial haplotypes. A particular haplotype almost absent in the North Sea was the most abundant in Belt Sea and Inner Baltic Sea. Microsatellites yielded a similar pattern (i.e., turnover in occurrence of clusters identified by STRUCTURE). Moreover, a highly significant association between microsatellite assignment and unlinked mitochondrial haplotypes further indicates a split between North Sea and Baltic porpoises. For the Inner Baltic Sea, we consistently recovered a small, but significant separation from the Belt Sea population. Despite recent arguments that separation should exceed a predefined threshold before populations shall be managed separately, we argue in favour of precautionary acknowledging the Inner Baltic porpoises as a separate management unit, which should receive particular attention, as it is threatened by various factors, in particular local fishery measures.

Reprint of: Temporal trends of polyfluoroalkyl compounds (PFCs) in liver tissue of grey seals (Halichoerus grypus) from the Baltic Sea, 1974-2008.

Temporal trends of polyfluoroalkyl compounds (PFCs) were examined in grey seal (Halichoerus grypus) liver from the Baltic Sea over a period of 35 years (1974-2008). In total, 17 of 43 PFCs were found, including the perfluoroalkyl sulfonates (C(4)-C(10) PFSAs), perfluorooctanesulfinate (PFOSi), long chain perfluoroalkyl carboxylates (C(7)-C(14) PFCAs), and perfluoroalkyl sulfonamides (i.e., perfluorooctane sulfonamide (FOSA) and N-ethyl perfluorooctane sulfonamide (EtFOSA)), whereas saturated and unsaturated fluorotelomer carboxylates, shorter chain PFCAs and perfluoroalkyl phosphonic acids were not detected. Perfluorooctane sulfonate (PFOS) was the predominant compound (9.57-1444 ng g(-1) wet weight (ww)), followed by perfluorononanoate (PFNA, 0.47-109 ng g(-1) ww). C(6)-C(8) PFSAs, PFOSi and C(7)-C(13) PFCAs showed statistically significant increasing concentrations between 1974 and 1997, with a peak in 1997 and then decreased or levelled off (except for C(12) and C(13) PFCAs). FOSA had a different temporal trend with a maximum in 1989 followed by significant decreasing concentrations until 2008. Toxicological implications for grey seals are limited, but the maximal PFOS concentration found in this study was about 40 times lower than the predicted lowest observed effect concentrations (LOEC). The statistically significant decreasing concentrations or levelling off for several PFCs in the relative closed marine ecosystem of the Baltic Sea indicate a rapidly responding to reduced emissions to the marine environment. However, the high concentrations of PFOS and continuing increasing concentrations of the longer chain PFCAs (C(12)-C(14)) shows that further work on the reduction of environmental emissions of PFCs are necessary.

Mass mortality in harbour seals and harbour porpoises caused by an unknown pathogen

EPIDEMICS caused by phocine distemper virus (pdv) resulted in mass mortalities of European harbour seals ( Phoca vitulina ) in 1988 and 2002 ([Harkonen and others 2006][1]). A third epidemic started in June 2007; the gross pathological changes observed in seals that died in the 2007 outbreak were

The Tail Feathers of Osprey Nestlings (Pandion haliaetus L.) as Indicators of Change in Mercury Load in the Environment of Southern Sweden (1969-1998): A Case Study with a Note on the Simultaneous Intake of Selenium

Abstract The tail feathers of 104 osprey nestlings (Pandion haliaetus) from Lake Åsnen, southern Sweden, were analyzed for total mercury and selenium content. Concentrations of mercury in feathers from the western part of the lake fell during the period 1969–1998, when pollution from industrial mercury had decreased and a paper mill upstream of the lake was closed down in 1979. Nestlings from the eastern part of the lake had initially lower levels of mercury, predominantly from atmospheric fallout. The levels did not decrease during the period. The reason for the differences seems to be the limited water flow between the western and the eastern part of the lake. The selenium concentrations were constant over time and approximately the same in both parts of the lake. The total-Hg/Se ratio in the western part of the lake was 2.94, which is somewhat higher than the 1:1 molar ratio constant (2.54). This indicates that the bioaccumulated quantity of selenium which is thought to be protective for the organisms against the toxic effects of mercury, is somewhat insufficient to bind all body mercury.

Increasing Concentrations of Perfluoroalkyl Acids in Scandinavian Otters (Lutra lutra) between 1972 and 2011: A New Threat to the Otter Population?

Liver samples from 140 otters (Lutra lutra) from Sweden and Norway were analyzed for 10 perfluoroalkyl carboxylic acids (PFCAs; C6-C15), 4 perfluoroalkane sulfonic acids (PFSAs; C4,C6,C8,C10) and perfluorooctane sulfonamide (FOSA). Perfluorooctane sulfonic acid (PFOS) was the dominant compound accounting for approximately 80% of the fluorinated contaminants and showing concentrations up to 16 μg/g wet weight. Perfluorononanoic acid (PFNA) was the dominant PFCA (up to 640 ng/g wet weight) closely followed by the C10 and C11 homologues. A spatial comparison between otters from southwestern Norway, southern and northern Sweden sampled between 2005 and 2011 revealed that the samples from southern Sweden had generally the largest contaminant load, but two PFCAs and FOSA were higher concentrated in the Norwegian samples. A temporal trend study was performed on otters from southern Sweden collected between 1972 and 2011. Seven PFCAs (C8-C14), PFOS and perfluorodecane sulfonic acid (PFDS) showed significantly increasing trends with doubling times between 5.5 and 13 years. The PFCAs also showed significantly increasing trends over the period 2002 to 2011. These findings together with the exceptionally high liver concentrations of PFOS are of great concern for the Scandinavian otter populations.

Spatially Explicit Analysis of Genome-Wide SNPs Detects Subtle Population Structure in a Mobile Marine Mammal, the Harbor Porpoise.

The population structure of the highly mobile marine mammal, the harbor porpoise (Phocoena phocoena), in the Atlantic shelf waters follows a pattern of significant isolation-by-distance. The population structure of harbor porpoises from the Baltic Sea, which is connected with the North Sea through a series of basins separated by shallow underwater ridges, however, is more complex. Here, we investigated the population differentiation of harbor porpoises in European Seas with a special focus on the Baltic Sea and adjacent waters, using a population genomics approach. We used 2872 single nucleotide polymorphisms (SNPs), derived from double digest restriction-site associated DNA sequencing (ddRAD-seq), as well as 13 microsatellite loci and mitochondrial haplotypes for the same set of individuals. Spatial principal components analysis (sPCA), and Bayesian clustering on a subset of SNPs suggest three main groupings at the level of all studied regions: the Black Sea, the North Atlantic, and the Baltic Sea. Furthermore, we observed a distinct separation of the North Sea harbor porpoises from the Baltic Sea populations, and identified splits between porpoise populations within the Baltic Sea. We observed a notable distinction between the Belt Sea and the Inner Baltic Sea sub-regions. Improved delineation of harbor porpoise population assignments for the Baltic based on genomic evidence is important for conservation management of this endangered cetacean in threatened habitats, particularly in the Baltic Sea proper. In addition, we show that SNPs outperform microsatellite markers and demonstrate the utility of RAD-tags from a relatively small, opportunistically sampled cetacean sample set for population diversity and divergence analysis.

Comparison of PFASs contamination in the freshwater and terrestrial environments by analysis of eggs from osprey (Pandion haliaetus), tawny owl (Strix aluco), and common kestrel (Falco tinnunculus)

The level of PFAS (per- and polyfluorinated alkyl substances) contamination in freshwater and terrestrial Swedish environments in 2013/2014 was assessed by analyzing a range of perfluorinated alkyl acids, fluorotelomer acids, sulfonamides, sulfonamidoethanols and polyfluoralkyl phosphate diesters (diPAPs) in predator bird eggs. Stable isotopes ((13)C and (15)N) were analyzed to elucidate the dietary source. The tawny owl (Strix aluco, n=10) and common kestrel (Falco tinnunculus, n=40), two terrestrial species, and the osprey (Pandion haliaetus, n=30), a freshwater specie were included. In addition, a temporal trend (1997-2001, 2008-2009, 2013) in osprey was studied as well. The PFAS profile was dominated by perfluorooctane sulfonic acid (PFOS) in eggs from osprey and tawny owl, while for common kestrel perfluorinated carboxylic acids (∑PFCA) exceeded the level of PFOS. PFOS concentration in osprey eggs remained at the same level between 1997 and 2001 and 2013. For the long-chained PFCAs, there were a significant increase in concentrations in osprey eggs between 1997 and 2001 and 2008-2009. The levels of PFOS and PFCAs were about 10 and five times higher, respectively, in osprey compared to tawny owl and common kestrel. Evidence of direct exposure from PFCA precursor compounds to birds in both freshwater and terrestrial environment was observed. Low levels of diPAPs were detected in a few samples of osprey (<0.02-2.4ng/g) and common kestrel (<0.02-0.16ng/g) eggs, and 6:2 FTSA was detected in a majority of the osprey eggs (<6.3-52ng/g). One saturated telomer acid (7:3 FTCA), which is a transformation marker from precursor exposure, was detected in all species (<0.24-2.7ng/g). The (15)N data showed higher levels in osprey eggs compared to tawny owl and common kestrel, indicating that they feed on a 2-3 times higher trophic level. We conclude that ospreys are continuously exposed to PFAS at levels where adverse toxic effects have been observed in birds.

Population structure and recent temporal changes in genetic variation in Eurasian otters from Sweden

The Eurasian otter (Lutra lutra) population in Sweden went through a drastic decline in population size between the 1950s and 1980s, caused mostly by anthropogenic factors such as high hunting pressure and the introduction of environmental toxic chemicals into the otter’s habitats. However, after the bans of PCBs and DDT in the 1970s, the population began to recover in the 1990s. This study compares microsatellite data across twelve loci from historical and contemporary otter samples to investigate whether there has been a change in population structure and genetic diversity across time in various locations throughout Sweden. The results suggest that otters in the south were more severely affected by the bottleneck, demonstrated by a decline in genetic diversity and a shift in genetic composition. In contrast, the genetic composition in otters from northern Sweden remained mostly unchanged, both in terms of population structure and diversity. This suggests that the decline was not uniform across the country. Moreover, our analyses of historical samples provide an overview of the level of genetic variation and population structure that existed prior to the bottleneck, which may be helpful for the future management and conservation of the species.

Temporal trends of PBDD/Fs, PCDD/Fs, PBDEs and PCBs in ringed seals from the Baltic Sea (Pusa hispida botnica) between 1974 and 2015

Temporal trends in exposure to persistent organic pollutants (POPs) were assessed in 22 pooled samples gathered from 69 individuals of Baltic ringed seal (Pusa hispida botnica) from 1974 to 2015. Samples were analysed for polybrominated dibenzo-p-dioxins and furans (PBDD/Fs), polychlorinated dibenzo-p-dioxins and furans (PCDD/Fs), polybrominated diphenyl ethers (PBDEs) and polychlorinated biphenyls (PCBs). No previous study has reported on the occurrence of PBDD/Fs in marine mammals in the Baltic Sea. Concentrations of pollutants in Baltic ringed seal, a marine mammal and top predator, can be used as an indicator of pollutants concentrations in the Baltic region. Visual inspection of data did not show any temporal trends for PBDD/Fs, while the PCDD/Fs and PCBs showed decreasing concentrations between 1974 and 2015. PBDEs increased until the end of the 1990s and then decreased until the end of the period. ∑PBDD/Fs ranged from 0.5-52.3pg/g lipid weight (l.w.) (0.08-4.8pgTEQ/g l.w.), with 1,2,3,4,6,7,8-HpBDF contributing on average 61% to ∑PBDD/Fs. ∑PCDD/Fs ranged from 103 to 1480pg/g l.w. (39-784pgTEQ/g l.w.), with 1,2,3,6,7,8-HxCDD, 1,2,3,7,8-PeCDD and 2,3,4,7,8-PeCDF showing the highest average concentrations. PBDD/F toxic equivalents (TEQ) contributed on average 1.1% to the total (PBDD/F+PCDD/F) TEQ. The ∑PBDEs concentration range was 18.7-503ng/g l.w., with BDE #47 the predominant congener. The concentration range for ∑PCBs was 2.8-40.1μg/g l.w., with #138 and #153 the most abundant congeners. Visual inspection of the data showed decreasing concentrations for all compound groups except PBDD/Fs. A slight increase in the PBDD/Fs concentrations was observed from 2004 onwards. This observation needs to be investigated further.

Distribution and molecular phylogeny of biliary trematodes (Opisthorchiidae) infecting native Lutra lutra and alien Neovison vison across Europe

The recent identification of Pseudamphistomum truncatum, (Rudolphi, 1819) (Trematoda: Opisthorchiidae) and Metorchis bilis (Braun, 1790) Odening, 1962 (synonymous with Metorchis albidus (Braun, 1893) Loos, 1899 and Metorchis crassiusculus (Rudolphi, 1809) Looss, 1899 (Trematoda: Opisthorchiidae)) in otters from Britain caused concern because of associated biliary damage, coupled with speculation over their alien status. Here, we investigate the presence, intensity and phylogeny of these trematodes in mustelids (principally otters) across Europe (Czech Republic, Denmark, France, Germany, Norway, Poland and Sweden and Britain). The trematodes were identified to species using the internal transcribed spacer II (ITS2) locus. Both parasites were found across Europe but at unequal frequency. In the German state of Saxony, eight out of eleven (73%) otters examined were infected with P. truncatum whilst this parasite was not found in either mink from Scotland (n=40) or otters from Norway (n=21). Differences in the phylogenies between the two species suggest divergent demographic histories possibly reflecting contrasting host diet or competitive exclusion, with M. bilis exhibiting greater mitochondrial diversity than P. truncatum. Shared haplotypes within the ranges of both parasite species probably reflect relatively unrestricted movements (both natural and anthropogenic) of intermediate and definitive hosts across Europe.