Anders Galatius

Investigating the Potential Use of Environmental DNA (eDNA) for Genetic Monitoring of Marine Mammals

The exploitation of non-invasive samples has been widely used in genetic monitoring of terrestrial species. In aquatic ecosystems, non-invasive samples such as feces, shed hair or skin, are less accessible. However, the use of environmental DNA (eDNA) has recently been shown to be an effective tool for genetic monitoring of species presence in freshwater ecosystems. Detecting species in the marine environment using eDNA potentially offers a greater challenge due to the greater dilution, amount of mixing and salinity compared with most freshwater ecosystems. To determine the potential use of eDNA for genetic monitoring we used specific primers that amplify short mitochondrial DNA sequences to detect the presence of a marine mammal, the harbor porpoise, Phocoena phocoena, in a controlled environment and in natural marine locations. The reliability of the genetic detections was investigated by comparing with detections of harbor porpoise echolocation clicks by static acoustic monitoring devices. While we were able to consistently genetically detect the target species under controlled conditions, the results from natural locations were less consistent and detection by eDNA was less successful than acoustic detections. However, at one site we detected long-finned pilot whale, Globicephala melas, a species rarely sighted in the Baltic. Therefore, with optimization aimed towards processing larger volumes of seawater this method has the potential to compliment current visual and acoustic methods of species detection of marine mammals.

Interspecific variation of ontogeny and skull shape among porpoises (Phocoenidae)

All extant members of Phocoenidae (porpoises) have been characterized as pedomorphic based on skeletal characters. To investigate the ontogenetic background for pedomorphosis and assess interspecific differences in ontogeny among phocoenids, samples of the six extant species were compared in terms of development of both epiphyseal and cranial suture fusion. Across all species, full maturity of the vertebral column was rare. Vertebral epiphyseal development did not progress so far in most Phocoena phocoena as in Phocoenoides dalli and Phocoena dioptrica. P. phocoena, Phocoena spinipinnis, Ph. dalli, and P. dioptrica, for which large series were available, were further compared in terms of ontogeny of cranial shape by three‐dimensional geometric morphometrics. Ph. dalli and P. dioptrica generally showed further development of cranial sutures than the other species. Postnatal skull shape development was similar for all species studied; the majority of interspecific shape differences are present at parturition. Smaller species had a higher rate of shape development relative to growth in size than Ph. dalli and P. dioptrica, but they still showed less allometric development due to less postnatal growth. Interspecific shape differences indicate phylogenetic relationships similar to that proposed based on morphology or convergent evolution of the two pelagic species, Ph. dalli and P. dioptrica, under the scenarios suggested by recent molecular studies. A shape trend coinciding with habitat preference was detected; in species with pelagic preference the position and orientation of the foramen magnum aligned the skull with the vertebral column; the rostrum showed less ventral inclination, and the facial region was larger and more concave in lateral aspect. J. Morphol., 2011.

Feeding behaviour of free-ranging walruses with notes on apparent dextrality of flipper use.

BackgroundDirect observations of underwater behaviour of free-living marine mammals are rare. This is particularly true for large and potentially dangerous species such as the walrus (Odobenus rosmarus). Walruses are highly specialised predators on benthic invertebrates – especially bivalves. The unique feeding niche of walruses has led to speculations as to their underwater foraging behaviour. Based on observations of walruses in captivity and signs of predation left on the sea floor by free-living walruses, various types of feeding behaviour have been suggested in the literature. In this study, however, the underwater feeding behaviour of wild adult male Atlantic walruses (O. r. rosmarus) is documented for the first time in their natural habitat by scuba-divers. The video recordings indicated a predisposition for use of the right front flipper during feeding. This tendency towards dextrality was explored further by examining a museum collection of extremities of walrus skeletons.ResultsDuring July and August 2001, twelve video-recordings of foraging adult male walruses were made in Young Sound (74°18 N; 20°15 V), Northeast Greenland. The recordings did not allow for differentiation among animals, however based on notes by the photographer at least five different individuals were involved. The walruses showed four different foraging behaviours; removing sediment by beating the right flipper, removing sediment by beating the left flipper, removing sediment by use of a water-jet from the mouth and rooting through sediment with the muzzle. There was a significant preference for using right flipper over left flipper during foraging. Measurements of the dimensions of forelimbs from 23 walrus skeletons revealed that the length of the right scapula, humerus, and ulna was significantly greater than that of the left, supporting our field observations of walruses showing a tendency of dextrality in flipper use.ConclusionWe suggest that the four feeding behaviours observed are typical of walruses in general, although walruses in other parts of their range may have evolved other types of feeding behaviour. While based on small sample sizes both the underwater observations and skeletal measurements suggest lateralized limb use, which is the first time this has been reported in a pinniped.

Sexually dimorphic proportions of the harbour porpoise (Phocoena phocoena) skeleton

Sexual differences in growth, allometric growth patterns and skeletal proportions were investigated by linear measurements of skeletal parts on 225 harbour porpoises (Phocoena phocoena) from the inner Danish and adjacent waters. Females show larger asymptotic sizes and extended period of growth compared with males. Measurements of the skull and flipper bones show negative allometry, whereas those of the bones of the body generally show positive allometry. There are no statistically significant intersexual differences in allometry except for the pelvic bones, where the males show stronger positive allometry. Throughout the range of individual sizes, females have significantly larger skulls and shorter vertebral columns than males for similarly sized individuals. In fully grown specimens, the condylobasal length of females makes up a smaller proportion of total length, and the vertebrae make up a larger proportion as compared with males. As these characters show negative and positive allometry, respectively, it is suggested that males finish their development at an earlier stage than females, retaining more paedomorphic proportions of the skeleton. Paedomorphosis in fully grown males relative to females is also found in the vertebral epiphyses that mature later in males than females, although the males finish growth at a younger age.

INCREASED CONTRACAECUM OSCULATUM INFECTION IN BALTIC COD (GADUS MORHUA) LIVERS (1982–2012) ASSOCIATED WITH INCREASING GREY SEAL (HALICHOERUS GRYPHUS) POPULATIONS

Abstract Grey seals (Halichoerus gryphus), the main final host of the gastric parasitic nematode Contracaecum osculatum in the Baltic, have recently recolonized the southwestern Baltic Sea. This colonization could lead to an increase in prevalence and intensity of third-stage larvae of C. osculatum in livers of Baltic cod (Gadus morhua), which serve as transport host for this helminth. We performed a parasitologic study of cod in spring 2012 and compared the results with previously unpublished data from 1982/1983. Additionally, grey seals were counted annually from 2000 to 2011 at three haul-out sites in the southwestern Baltic. Of 97 cod livers examined in the 1982/1983 survey, 22% harbored C. osculatum larvae, whereas 55.1% of the examined cod livers (n = 185) were infected in 2012; the mean intensity and mean abundance increased from 4.3 and 0.9 to 20.2 and 11.1, respectively. Molecular identification (PCR) confirmed the identity of the larvae. The grey seal population increased markedly during the 12-yr period. We suggest that the elevated parasitism of cod livers is associated with the successful re-establishment of grey seals in the southwestern Baltic.

Population structure of harbour porpoises in the Baltic region: evidence of separation based on geometric morphometric comparisons

The harbour porpoise is seriously depleted and threatened with extinction in the Baltic Sea. It is usually assumed that Baltic porpoises form a separate population unit, although the evidence for this has been disputed lately. Here, a 3-D geometric morphometric approach was employed to test a number of hypotheses regarding population structure of the harbour porpoise in the Baltic region. 277 porpoise skulls from Denmark, Sweden, Finland, Germany and Poland were measured with a suite of 3-D landmarks. Statistical analyses revealed highly significant shape differences between porpoises from the North Sea, Belt Sea and the inner Baltic Sea. A comparison of the directionalities of the shape vectors between these units found differences that cannot be attributed to a general, continual shape trend going from the North Sea to the inner Baltic Sea. These vectors indicate a morphological adaptation to the specific sub-areas. Such adaptation may be the result of the topographic peculiarities of the area with variable topography and shallow waters, e.g. in the Belt Sea porpoises, there may be a greater reliance on benthic and demersal prey. The present results show that isolation by distance alone is an unlikely explanation for the differences found within the Baltic region and thus support previously reported molecular indications of a separate population within the inner Baltic Sea.

Temporal and life history related trends of perfluorochemicals in harbor porpoises from the Danish North Sea

Eighty-five stranded or bycaught harbor porpoises collected from the Danish North Sea between 1980 and 2005 were analyzed for perfluorochemicals in the liver. PFOS was the predominant compound, making up on average 88.9% of the ∑PFC, followed by PFOSA (7.8%). PFUnA (1.9%) and PFDA (1.2%) were detected in most samples. PFHxS, PFNA and PFOA were only found in a minority of the samples. We found substantial differences in PFC concentrations among life history stages, the highest concentrations were found in neonates, suckling juveniles and lactating females. Such differences should be considered when PFC levels in wildlife are evaluated. The high concentrations found in young porpoises are of concern as PFCs have known toxic effects on the development of the central nervous system and reproductive organs. Despite efforts to reduce PFC emissions, a decreasing temporal trend of concentrations was not detected for any compound. PFCA concentrations were found to be increasing.

OCCURRENCE OF VERTEBRAL OSTEOPHYTOSIS IN A MUSEUM SAMPLE OF WHITE-BEAKED DOLPHINS (LAGENORHYNCHUS ALBIROSTRIS) FROM DANISH WATERS

The occurrence of pathologic new bone formation in the vertebral column was studied in 46 skeletons of the white-beaked dolphin (Lagenorhynchus albirostris) stranded in Denmark between 1903 and 2002 and held in the collections of the Zoological Museum, University of Copenhagen. New bone formation was evident in 18 (72%) of 25 physically mature specimens and in one of 21 physically immature specimens. This pattern of occurrence is consistent with spondylosis deformans, which is caused by degeneration of the intervertebral disk. In general, females seemed to be more susceptible, exhibiting a higher number of affected vertebrae and more severe cases. Lesions were predominantly detected in the lumbar and cranial caudal vertebra. Data on blubber thickness were available for part of the sample; one of the most heavily afflicted specimens had the thickest recorded blubber, indicating that secondary bone formation leading to blocks of fused vertebrae had not necessarily impaired the body condition of afflicted specimens. However, age determinations conducted on the sample revealed that none of the most severe cases occurred among the oldest animals, implying that these severe cases may cause premature fatality.

Spatial trends of perfluorochemicals in harbor seals (Phoca vitulina) from Danish waters

Spatial trends of concentrations of perfluorinated chemicals (PFCs) were investigated in harbour seal liver tissue from seven locations in Denmark, ranging from the Wadden Sea in the southern North Sea to the Western Baltic. All samples were collected during the phocine distemper epizootic in 2002 which provided access to a large number of comparable samples over a short time period. PFOS was dominating (mean: 92% of ∑PFC) among the PFCs in the samples, followed by considerably lower concentrations of PFHxS (1.8%), PFDA (1.7%), PFNA (1.6%) PFUnA (1.5%), PFOA (0.9%) and PFOSA (0.5%). The concentrations of all the investigated compounds showed significant differences among the seven locations. PFOS showed the highest concentrations in the Wadden Sea, where high burdens have also been recorded in German seals. Most compounds showed a trend towards higher concentrations at one or both extremes of the geographic range. Two different patterns of relative PFC concentrations were detected; one in the inner Danish waters where PFOSA and PFUnA were more prevalent and another in the Wadden Sea and Limfjord where PFOA, PFHxS and PFNA were found in greater proportions. These patterns probably represent Baltic and North Sea contamination sources.

Shift of grey seal subspecies boundaries in response to climate, culling and conservation

Identifying the processes that drive changes in the abundance and distribution of natural populations is a central theme in ecology and evolution. Many species of marine mammals have experienced dramatic changes in abundance and distribution due to climatic fluctuations and anthropogenic impacts. However, thanks to conservation efforts, some of these species have shown remarkable population recovery and are now recolonizing their former ranges. Here, we use zooarchaeological, demographic and genetic data to examine processes of colonization, local extinction and recolonization of the two northern European grey seal subspecies inhabiting the Baltic Sea and North Sea. The zooarchaeological and genetic data suggest that the two subspecies diverged shortly after the formation of the Baltic Sea approximately 4200 years bp, probably through a gradual shift to different breeding habitats and phenologies. By comparing genetic data from 19th century pre‐extinction material with that from seals currently recolonizing their past range, we observed a marked spatiotemporal shift in subspecies boundaries, with increasing encroachment of North Sea seals on areas previously occupied by the Baltic Sea subspecies. Further, both demographic and genetic data indicate that the two subspecies have begun to overlap geographically and are hybridizing in a narrow contact zone. Our findings provide new insights into the processes of colonization, extinction and recolonization and have important implications for the management of grey seals across northern Europe.