Gene expression and trace elements in Greenlandic ringed seals (Pusa hispida).

Marine top predators such as ringed seals biomagnify environmental contaminants; and with the increasing human activities in the Arctic, ringed seals are exposed to biologically significant concentrations of trace elements resulting in reproductive impairment, immunosuppression, and neurological damages. Little is known about the molecular effects of heavy metals on these vulnerable apex predators suffering from a rapidly changing Arctic with significant loss of sea-ice. In the present study, concentrations of cadmium (Cd), mercury (Hg) and selenium (Se) were measured in liver of sixteen Greenlandic ringed seals (nine adults and seven subadults) together with molecular biomarkers involved in bio-transformation, oxidative stress, endocrine disruption and immune activity in blood and blubber. The concentrations of trace elements increased in the following order: Hg > Se > Cd with levels of mercury and selenium being highest in adults. Aryl hydrocarbon receptor nuclear translocator (ARNT), peroxisome proliferator activated receptor alpha (PPARα, estrogen receptor alpha (ESR1), thyroid hormone receptor alpha (TRα) and interleukin - 2 (IL-2) mRNA transcript levels were highest in blubber, while heat shock protein 70 (HSP70) and interleukin - 10 (IL-10) were significantly higher in blood. There were no significant correlations between the concentrations of trace elements and mRNA transcript levels suggesting that stressors other than the trace elements investigated are responsible for the changes in gene expression levels. Since Hg seems to increase in Greenlandic ringed seals, there is a need to re-enforce health monitoring of this ringed seal population.

Heartworms in Halichoerus grypus: first records of Acanthocheilonema spirocauda (Onchocercidae; Filarioidea) in 2 grey seals from the North Sea.

The assumed definitive host of the heartworm Acanthocheilonema spirocauda (Onchocerdidae; Filarioidea) is the harbour seal (Phoca vitulina). This filaroid nematode parasitizing in cardiac ventricles and blood vessel lumina of harbour seals (P. vitulina) has a low prevalence and seldom causes severe health impacts. The seal louse (Echinophthirius horridus) is the assumed intermediate host for transmission of A. spirocauda filariae between seals, comprising a unique parasite assembly conveyed from the terrestrial ancestors of pinnipeds. Although grey seals (Halichoerus grypus) are infected by seal lice, heartworm infection was not verified. Analysing a longterm dataset compiled over decades (1996­2021) of health monitoring seals along the German coasts comprising post mortem investigations and archived parasites, 2 cases of A. spirocauda infected male grey seals were detected. Tentative morphological identification was confirmed with molecular tools by sequencing a section of mtDNA COI and comparing nucleotide data with available heartworm sequence. This is the first record of heartworm individuals collected from the heart of grey seals at necropsy. It remains puzzling why heartworm infection occur much less frequently in grey than in harbour seals, although both species use the same habitat, share mixed haul-outs and consume similar prey species. If transmission occurs directly via seal louse vectors on haul-outs, increasing seal populations in the North- and Baltic Sea could have density dependent effects on prevalence of heartworm and seal louse infections. It remains to be shown how species-specificity of filarial nematodes as well as immune system traits of grey seals influence infection patterns of A. spirocauda.

Deciphering Eurasian otter (Lutra lutra L.) and seal (Phoca vitulina L.; Halichoerus grypus F.) diet: Metabarcoding tailored for fresh and saltwater fish species.

Long-lived top predators shape biodiversity structure in their ecosystems and predator-prey interactions are critical in decoding how communities function. Studies on the foraging ecology of seals and Eurasian otters in Western Europe are outdated and most studies solely performed traditional hard part analysis. Molecular metabarcoding can be used as an innovative noninvasive diet analysis tool, which has proven efficient and complementary to hard part analysis, however, lacking application in the wider North Sea area. In this study, DNA from digesta, collected between 2014-2020, were used to identify fish species in the diet of 47 Eurasian otters, 54 harbour seals and 21 grey seals by applying a next-generation metabarcoding approach. A newly designed 16S rRNA primer, providing the best coverage of >130 local marine and freshwater fish species, was used to amplify prey DNA from seal scats and otter gut content sampled from the North Sea and regional freshwater bodies. Frequent fish species included tench, ninespine stickleback and white bream in otters; hooknose and common roach in grey seals and Pleuronectidae and sand gobies in harbour seals. Bipartite network analysis showed a strong overlap of harbour and grey seal diets. Otter diet intersected with both seal species in terms of freshwater species. This study provides new knowledge about dietary composition and community assemblage of fish prey in otters and seals in the North Sea and regional freshwaters, and a new molecular tool to elucidate predator-prey interactions and interspecies competition in complex and changing ecosystems under pressure from anthropogenic activities.

Novel infections of Corynosoma enhydri and Profilicollis sp. (Acanthocephala: Polymorphidae) identified in sea otters Enhydra lutris.

Infectious disease is a major cause of mortality for sea otters Enhydra lutris, a keystone species of continued concern for conservationists. Parasitic infection has long been identified as a cause of mortality in otters in both Alaska and California, USA. Corynosoma enhydri (Acanthocephala) is the only parasite that uses sea otters as its primary definitive host and is highly prevalent in otter populations; however, it is generally considered unimportant both pathologically and ecologically, although this assumption is based on limited empirical knowledge. Research has instead focused on Profilicollis infections (P. major, P. kenti, P. altmani) as a significant source of otter mortality due to associated enteritis and peritonitis, which are threats to otter health. Here we describe acanthocephalan infections in sea otters by Profilicollis spp. and C. enhydri, from a survey comparing C. enhydri infections between northern sea otters E. lutris kenyoni (n = 12) and southern sea otters E. lutris nereis (n = 19). We report a novel infection of C. enhydri in a pup approximately 1 mo of age, which shows that the early introduction to solid food at around 3 wk by their mothers may lead to subsequent infection via infected prey items. We also document the first 2 known cases of Profilicollis infection in northern sea otters, which may present an unknown threat to the Alaskan population, or may be an interesting example of accidental infection.

Pathological findings in North Sea and Baltic grey seal and harbour seal intestines associated with acanthocephalan infections.

Grey seals Halichoerus grypus and harbour seals Phoca vitulina are common seal species in the North and Baltic seas and final hosts of Corynosoma acanthocephalans. C. strumosum and C. magdaleni infect the small intestines of both seal species. In contrast to harbour seals, Baltic grey seals in the past have regularly displayed severe C. semerme infections in the caecum and colon, with associated tunica muscularis hypertrophy, inflammation and ulcerations as part of the Baltic seal disease complex (BSDC). Pathogenesis and correlation of acanthocephalan infections with these lesions are still unknown. This study describes the intestinal pathology and parasitic distribution in each seal species. Grey seal (n = 83) and harbour seal (n = 1156) intestines of all age groups and sexes, collected in Poland, Germany and Estonia from 1998 to 2017, were investigated. Most harbour seals came from the North Sea, whereas grey seals were predominantly derived from the Baltic Sea. Both species featured mild to moderate small intestinal infections. Grey seals showed colonic infections not found in harbour seals and featured a chronic erosive to ulcerative, eosinophilic or lympho-plasmacytic colitis with tunica muscularis hypertrophy, indicating still prevailing clinical signs of the BSDC. Harbour seals displayed granulomatous, eosinophilic, lympho-plasmacytic or catarrhal enteritis. The prevalence of acanthocephalan infections in harbour seals increased from 2012 onwards. Furthermore, significant associations between acanthocephalan infection and the presence of intestinal inflammation were found for both seal species. This study suggests that the level of acanthocephalan infection and associated lesions are suitable seal population health indicators, with the colon being a specific target organ for Baltic grey seal health monitoring.

Phylogenomic insights to the origin and spread of phocine distemper virus in European harbour seals in 1988 and 2002.

The 1988 and 2002 phocine distemper virus (PDV) outbreaks in European harbour seals Phoca vitulina are among the largest mass mortality events recorded in marine mammals. Despite its large impact on harbour seal population numbers, and 3 decades of studies, many questions regarding the spread and temporal origin of PDV remain unanswered. Here, we sequenced and analysed 7123 bp of the PDV genome, including the coding and non-coding regions of the entire P, M, F and H genes in tissues from 44 harbour seals to shed new light on the origin and spread of PDV in 1988 and 2002. The phylogenetic analyses trace the origin of the PDV strain causing the 1988 outbreak to between May 1987 and April 1988, while the origin of the strain causing the 2002 outbreak can be traced back to between June 2001 and May 2002. The analyses further point to several independent introductions of PDV in 1988, possibly linked to a southward mass immigration of harp seals in the winter and spring of 1987-1988. The vector for the 2002 outbreak is unknown, but the epidemiological analyses suggest the subsequent spread of PDV from the epicentre in the Kattegat, Denmark, to haul-out sites in the North Sea through several independent introductions.

CRYPTOSPORIDIUM SPECIES AND GIARDIA SPECIES PREVALENCE IN MARINE MAMMAL SPECIES PRESENT IN THE GERMAN NORTH AND BALTIC SEAS.

Marine mammals are known to harbor Cryptosporidium spp. and Giardia spp., especially species or genotypes commonly associated with humans. In animals of the North and Baltic Seas, this information is missing. In this study, fecal samples ( n = 97) from harbor porpoises ( Phocoena phocoena), harbor seals ( Phoca vitulina), and grey seals ( Phocoena phocoena) present in German waters were examined via microscopic examination (by using a modified Ziehl-Neelsen technique [MZN]) to estimate the prevalence of these parasites. Additionally, intestinal tissue samples ( n = 1,049) from specimens of 10 marine mammal species were submitted for histopathologic examination (HP). A low prevalence was detected (2.1 and 1.3%, for MZN and HP, respectively), associated with Cryptosporidium spp. Giardia spp. was not observed. Current results suggest a low prevalence of these parasites in marine mammals present in Germany. However, methods with higher sensitivity, such as molecular methods, are crucial to confirm the prevalence, establish origin routes, and determine epidemiologic factors associated with the low prevalence observed.

Parasites in the inner ear of harbour porpoise: cases from the North and Baltic Seas.

Peribullar sinuses of harbour porpoises Phocoena phocoena are parasitized with high prevalence by the nematode Stenurus minor. The effect of S. minor on the hearing ability of this species is still undetermined. Here, we review the occurrence of S. minor in the inner ear of harbour porpoises recovered from strandings in the North and Baltic Seas. In particular, we present the results from ears collected in German and Danish waters from 2002 to 2016 and from Dutch waters from 2010 to 2016. While the prevalence of S. minor in pterygoid and peribullar sinuses and tympanic cavity was high in harbour porpoises (66.67% in our cases), its prevalence in the cochlea was rare. Only 1 case out of 129 analysed by either histology, electron microscopy or immunofluorescence showed the presence of a nematode parasite morphologically consistent with S. minor at the most basal portion of the right cochlea. This individual also had severe haemorrhage along the right cochlear spiral, which was likely caused by ectopic S. minor migration. Although this animal might have had impaired hearing in the right ear, it was otherwise in good body condition with evidence of recent feeding. These findings highlight the need to study the effect of parasites on hearing, and other pathological changes that might impair appropriate processing of acoustic information.

Metazoan parasites from odontocetes off New Zealand: new records.

Information about the parasite fauna of spectacled porpoises and cetaceans from New Zealand waters in general is scarce. This study takes advantage of material archived in collections of the Otago Museum in Dunedin and Massey University in Auckland, sampled from cetacean species found stranded along the New Zealand coastline between 2007 and 2014. Parasites from seven species of cetaceans (spectacled porpoise, Phocoena dioptrica (n = 2 individuals examined); pygmy sperm whale (n = 1); long-finned pilot whale, Globicephala melas (n = 1); Risso's dolphin, Grampus griseus (n = 1); short-beaked common dolphin, Delphinus delphis (n = 7); striped dolphin, Stenella coeruleoalba (n = 3) and dusky dolphin, Lagenorhynchus obscurus (n = 2)) from the respiratory and gastro-intestinal tract, cranial sinus, liver, urogenital and mammary tract, fascia and blubber were investigated. Ten parasite species were identified, belonging to the Nematoda (Stenurus minor, Stenurus globicephalae, Halocercus sp. (Pseudaliidae), Anisakis sp. (Anisakidae), Crassicauda sp. (Crassicaudidae)), Cestoda (Phyllobothrium delphini and Monorygma grimaldii (Phyllobothriidae)), Trematoda (Brachicladium palliata and Brachicladium delphini (Brachicladiidae)) and Crustacea (Scutocyamus antipodensis (Cyamidae)). Some of the parasite species encountered comprises new records for their host. Although the material was not sampled within a systematic parasitological survey, the findings contain valuable new information about the parasite fauna of rare, vagile and vulnerable marine wildlife from a remote oceanic environment.

Impacts of Underwater Noise on Marine Vertebrates: Project Introduction and First Results.

The project conducts application-oriented research on impacts of underwater noise on marine vertebrates in the North and Baltic Seas. In distinct subprojects, the hearing sensitivity of harbor porpoises and gray seals as well as the acoustic tolerance limit of harbor porpoises to impulsive noise from pile driving and stress reactions caused by anthropogenic noise is investigated. Animals are equipped with DTAGs capable of recording the actual surrounding noise field of free-swimming harbor porpoises and seals. Acoustic noise mapping including porpoise detectors in the Natura 2000 sites of the North and Baltic Seas will help to fully understand current noise impacts.

Xenobiotic and Immune-Relevant Molecular Biomarkers in Harbor Seals as Proxies for Pollutant Burden and Effects.

Harbor seals are exposed to increasing pressure caused by anthropogenic activities in their marine environment. Persistent organic pollutants (POPs) and trace elements are hazardous contaminants that accumulate in tissues of harbor seals. POPs and trace elements can negatively affect the immune-system and have been reported, e.g., to increase susceptibility to viral infections in seals. Biomarkers of the xenobiotic metabolism, cytokines, and heat-shock protein as cell mediators of the immune-system were established to evaluate the impact of environmental stressors on harbor seals. Harbor seals (n = 54) were captured on sandbanks in the North Sea during 2009-2012. Health assessments, including hematology, were performed, and RNAlater blood samples were taken and analyzed using quantitative polymerase chain reaction. Normalized transcript copy numbers were correlated to hematology and POP concentration in blood and trace metals in blood and fur. A significant correlation between xenobiotic markers and contaminant burden was found. Significant interrelationships between markers and POP compounds, as well as with season, weight, and hematology values, indicate that biomarkers reflect pollutant exposure and effects. A significant relationship between cortisol levels and heat-shock protein expression was observed indicating stress experienced during restraint of the seals. Interleukin-10 transcription showed significant correlations with trace elements in fur pointing toward immune regulatory effects of metal exposure. The molecular markers prove to be an important noninvasive tool that reflects contaminant exposure and the impact of anthropogenic stressors in seal species. The connection between interleukin-2, xenobiotic markers, and pollutants may indicate immune suppression in animals exposed to contaminants with subsequent susceptibility to inflammatory disease.

High prevalence and low intensity of Echinophthirius horridus infection in seals revealed by high effort sampling.

Seal lice (Echinophthirius horridus) are bloodsucking ectoparasites of phocid seals and vectors of pathogens like the heartworm, Acanthocheilonema spirocauda. Grey and harbour seal populations are recovering in German waters and wildlife health surveillance is crucial for wildlife conservation. A new, high effort sampling protocol for seal lice was applied for grey and harbour seals along the German North- and Baltic Sea coast. Freshly dead seals were systematically sampled within a health monitoring of stranded seals over 12 months. Prevalence, intensity and distribution patterns of seal lice were analysed. 58% of harbour seals (n = 71) and 70% of grey seals (n = 10) were infected with seal lice. A majority of harbour seals displayed mild levels of infection, while three were moderately and two were severely infected. The head was the preferred predilection site, indicating that E. horridus prefers body areas with frequent access to atmospheric oxygen. Nits and different developmental stages were recorded in all age classes in grey and harbour seals in all seasons. For the first time, copulating specimens of E. horridus were recorded on a dead harbour seal, highlighting that E. horridus reproduces throughout the year on seals of all age classes in German waters.

Attachment performance of the ectoparasitic seal louse Echinophthirius horridus.

Marine mammals host a great variety of parasites, which usually co-evolved in evolutionary arms races. However, little is known about the biology of marine mammal insect parasites, and even less about physical aspects of their life in such a challenging environment. One of 13 insect species that manage to endure long diving periods in the open sea is the seal louse, Echinophthirius horridus, parasitising true seals. Its survival depends on its specialised adaptations for enduring extreme conditions such as hypoxia, temperature changes, hydrostatic pressure, and strong drag forces during host dives. To maintain a grip on the seal fur, the louse's leg morphology is equipped with modified snap hook claws and soft pad-like structures that enhance friction. Through techniques including CLSM, SEM, and histological staining, we have examined the attachment system's detailed structure. Remarkably, the seal louse achieves exceptional attachment forces on seal fur, with safety factors (force per body weight) reaching 4500 in average measurements and up to 18000 in peak values, indicating superior attachment performance compared to other insect attachment systems. These findings underscore the louse's remarkable adaptations for life in a challenging marine environment, shedding light on the relationship between structure and function in extreme ecological niches.

Heartworm and seal louse: Trends in prevalence, characterisation of impact and transmission pathways in a unique parasite assembly on seals in the North and Baltic Sea.

The ectoparasitic seal louse, Echinophthirius horridus infects harbour (Phoca vitulina) and grey seals (Halichoerus grypus) in the North and Baltic Sea. The endoparasitic heartworm Acanthocheilonema spirocauda parasitizes the right heart and blood vessels of harbour seals. The complete lifecycle of the heartworm is not entirely understood although the seal louse is assumed to serve as vector for its transmission. Knowledge about the impact of both parasite species on host health are scarce. In this study, necropsy data and archived parasites of harbour and grey seals in German waters were analysed to determine long-term seal louse (SLP) and heartworm prevalence (HWP) from 2014 to 2021. Histology, microbiology and scanning electron microscopy (SEM) were applied on seal louse infected and uninfected skin to investigate associated lesions and the health impact. During the study period, HWP in harbour seals was 13%, the SLP in harbour seals was 4% and in grey seals 10%. HWP of harbour seals was significantly higher during the winter months compared to the summer. SLP in adults was significantly higher in comparison to juvenile harbour seals. SLP varied significantly between grey seals from the North and Baltic Sea. Filarial nematodes were detected in the haemocoel, pharynx, and intestine of E. horridus highlighting the seal louse as vector for heartworms. Alopecia and folliculitis were associated with the attachment posture of E. horridus and microbiological investigations isolated bacteria commonly associated with folliculitis.

Evaluation of immune and stress status in harbour porpoises (Phocoena phocoena): can hormones and mRNA expression levels serve as indicators to assess stress?

BACKGROUND: The harbour porpoise is exposed to increasing pressure caused by anthropogenic activities in its marine environment. Numerous offshore wind farms are planned or under construction in the North and Baltic Seas, which will increase underwater noise during both construction and operation. A better understanding of how anthropogenic impacts affect the behaviour, health, endocrinology, immunology and physiology of the animals is thus needed. The present study compares levels of stress hormones and mRNA expression of cytokines and acute-phase proteins in blood samples of harbour porpoises exposed to different levels of stress during handling, in rehabilitation or permanent human care.Free-ranging harbour porpoises, incidentally caught in pound nets in Denmark, were compared to harbour porpoises in rehabilitation at SOS Dolfijn in Harderwijk, the Netherlands, and individuals permanently kept in human care in the Dolfinarium Harderwijk and Fjord & Belt Kerteminde, Denmark. Blood samples were investigated for catecholamines, adrenaline, noradrenaline and dopamine, as well as for adrenocorticotropic hormone (ACTH), cortisol, metanephrine and normetanephrine. mRNA expression levels of relevant cell mediators (cytokines IL-10 and TNFα, acute-phase proteins haptoglobin and C-reactive protein and the heat shock protein HSP70) were measured using real-time PCR. RESULTS: Biomarker expression levels varied between free-ranging animals and porpoises in human care. Hormone and cytokine ranges showed correlations to each other and to the health status of investigated harbour porpoises. Hormone concentrations were higher in free-ranging harbour porpoises than in animals in human care. Adrenaline can be used as a parameter for the initial reaction to acute stress situations; noradrenaline, dopamine, ACTH and cortisol are more likely indicators for the following minutes of acute stress. There is evidence for different correlations between production of normetanephrine, metanephrine, cortisol and the expression of IL-10, HSP70 and haptoglobin. CONCLUSIONS: The expression patterns of the selected molecular biomarkers of the immune system are promising to reflect the health and immune status of the harbour porpoise under different levels of stress.

Potential new species of pseudaliid lung nematode (Metastrongyloidea) from two stranded neonatal orcas (Orcinus orca) characterized by ITS-2 and COI sequences.

Knowledge about parasite species of orcas, their prevalence, and impact on the health status is scarce. Only two records of lungworm infections in orca exist from male neonatal orcas stranded in Germany and Norway. The nematodes were identified as Halocercus sp. (Pseudaliidae), which have been described in the respiratory tract of multiple odontocete species, but morphological identification to species level remained impossible due to the fragile structure and ambiguous morphological features. Pseudaliid nematodes (Metastrongyloidea) are specific to the respiratory tract of toothed whales and are hypothesized to have become almost extinct in terrestrial mammals. Severe lungworm infections can cause secondary bacterial infections and bronchopneumonia and are a common cause of mortality in odontocetes. DNA isolations and subsequent sequencing of the rDNA ITS-2 and mtDNA COI revealed nucleotide differences between previously described Halocercus species from common dolphin (H. delphini) and harbor porpoises (H. invaginatus) that were comparatively analyzed, pointing toward a potentially new species of pseudaliid lungworm in orcas. New COI sequences of six additional metastrongyloid lungworms of seals and porpoises were derived to elucidate phylogenetic relationships and differences between nine species of Metastrongyloidea.

Back from Exile? First Records of Chewing Lice (Lutridia exilis; Ischnocera; Mallophaga) in Growing Eurasian Otter (Lutra lutra) Populations from Northern Germany.

Arthropod ectoparasites of aquatic wildlife often have complex relationships with their host species that have developed over long evolutionary time scales. Specialist parasite occurrence might depend on these hosts' distributions. Eurasian otter (Lutra lutra) populations are recovering in Northern German federal states, such as Schleswig-Holstein and Lower Saxony. Chewing lice (Lutridia exilis; Ischnocera; Mallophaga) are considered otter-specific yet rare parasites in their known range. In 2022, they were recorded for the first time on nine otters found dead in Northern Germany. All otters originated from the years 2021-2022 and were dissected during population health monitoring programs in 2022. Females (n = 6) were 0-5.5 years old and showed signs of disease in five cases. Males (n = 3), in contrast, were 0-1.6 years old and showed disease in a single case. Individual lice intensity of infection ranged from 1 to 75 specimens per otter. No direct adverse health effects of chewing lice on the otters were noted. Lutridia exilis morphological characteristics were documented and measurements were taken to study specialized adaptations that allow lice to attach to semi-aquatic otters. In addition, morphology was compared between lice from different geographical regions and specimens from previous reports. A region of the COI mDNA was amplified to molecularly characterize L. exilis for the first time and detect genetic differences between otter lice populations in Germany. It is believed that specialist parasites reduce in numbers even before their host populations decline. Recovering otter populations in Northern Germany could be an example of a reverse effect, where the comeback of a host species results in the return of a specialist parasite, which reflects an ultimate boost in overall species biodiversity.

Variation of blubber thickness for three marine mammal species in the southern Baltic Sea.

Evaluating populational trends of health condition has become an important topic for marine mammal populations under the Marine Strategy Framework Directive (MSFD). In the Baltic Sea, under the recommendation of Helsinki Commission (HELCOM), efforts have been undertaken to use blubber thickness as an indicator of energy reserves in marine mammals. Current values lack geographical representation from the entire Baltic Sea area and a large dataset is only available for grey seals (Halichoerus grypus) from Sweden and Finland. Knowledge on variation of blubber thickness related to geography throughout the Baltic Sea is important for its usage as an indicator. Such evaluation can provide important information about the energy reserves, and hence, food availability. It is expected that methodological standardization under HELCOM should include relevant datasets with good geographical coverage that can also account for natural variability in the resident marine mammal populations. In this study, seasonal and temporal trends of blubber thickness were evaluated for three marine mammal species-harbor seal (Phoca vitulina), grey seal (Halichoerus grypus) and harbor porpoise (Phocoena phocoena)-resident in the southern Baltic Sea collected and investigated under stranding networks. Additionally, the effects of age, season and sex were analyzed. Seasonal variation of blubber thickness was evident for all species, with harbor seals presenting more pronounced effects in adults and grey seals and harbor porpoises presenting more pronounced effects in juveniles. For harbor seals and porpoises, fluctuations were present over the years included in the analysis. In the seal species, blubber thickness values were generally higher in males. In harbor seals and porpoises, blubber thickness values differed between the age classes: while adult harbor seals displayed thicker blubber layers than juveniles, the opposite was observed for harbor porpoises. Furthermore, while an important initial screening tool, blubber thickness assessment cannot be considered a valid methodology for overall health assessment in marine mammals and should be complemented with data on specific health parameters developed for each species.

Preliminary study of oxidative stress biomarkers and trace elements in North Sea Harbour Seals.

This preliminary study investigated the potential correlations between trace elements (mercury, zinc, cadmium, copper, selenium, lead, nickel, chromium, lithium and vanadium) concentrations, measured in red blood cells, and oxidative stress biomarkers (total thiols, total glutathione, total and selenium-dependent glutathione peroxidases, triglycerides, malondialdehyde) assessed in the respective serum, in males and females P. vitulina, sampled in the Wadden Sea in spring and autumn 2015. Only concentrations of total mercury and zinc showed significant differences by sex, and only lipid peroxidation was different by season. Moreover, significant positive and negative correlations were observed between biomarkers (triglycerides, thiols, malondialdehyde, glutathione) and trace element concentrations (copper, lead, mercury, nickel, zinc). These findings suggest that the studied biomarkers could be useful for the assessment of oxidative stress in harbour seals exposed to trace elements, but further research with larger sample sizes is needed to better understand their specific associations.

Lungworm infections in harbour porpoises (Phocoena phocoena) in the German Wadden Sea between 2006 and 2018, and serodiagnostic tests.

Pseudaliid lungworm (Metastrongyloidea) infections and associated secondary bacterial infections may severely affect the health status of harbour porpoises (Phocoena phocoena) in German waters. The presented retrospective analysis including data from 259 harbour porpoises stranded between 2006 and 2018 on the German federal state of Schleswig-Holstein's North Sea coast showed that 118 (46%) of these stranded individuals harboured a lungworm infection. During this 13-year period, a significant difference in annual lungworm prevalence was only observed between the years 2006 and 2016. Lungworm coinfections of bronchi and pulmonary blood vessels were observed in 85.6% of positive cases. Mild infection levels were detected in 22.9% of infected animals and were most common in the age class of immature individuals (74.1%). Moderate and severe infections were present in 38.1% and 39.0% of the lungworm positive animals, respectively. Their distribution in immatures (51.1% and 54.3%) and adults (48.9% and 43.4%) did not show significant differences. In stranded animals, lungworm diagnosis can be easily obtained via necropsy, while reliable lungworm diagnosis in living porpoises requires invasive bronchoscopy or faecal examination, which is difficult to obtain in cetaceans. To overcome this issue, an enzyme-linked immunosorbent assay (ELISA) and immunoblot based on recombinant major sperm protein (MSP) of the cattle lungworm were evaluated as potential diagnostic tools in harbour porpoises. However, in contrast to hitherto other investigated host species, no reliable antibody response pattern was detectable in harbour porpoise serum/plasma or whole blood samples. Thus, MSP-based serological tests are considered unsuitable for lungworm diagnosis in harbour porpoises.