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Traditional bulk isotopic analysis is a pivotal tool for mapping consumer–resource interactions in food webs but has largely failed to adequately describe parasite–host relationships. Thus, parasite–host interactions remain undescribed in food web frameworks despite these relationships increasing linkage density, connectance and ecosystem biomass. Compound-specific stable isotopes from amino acids provides a promising novel approach that may aid in mapping parasite–host relationships in food webs. Here we apply a combination of traditional bulk stable isotope analyses and compound-specific isotopic analysis of nitrogen in amino acids to examine resource use and trophic interactions of five parasites from three hosts from a marine coastal food web (Wadden Sea, European Atlantic). By comparing isotopic compositions of bulk and amino acid nitrogen, we aimed to characterize isotopic fractionation occurring between parasites and their hosts and to clarify parasite trophic positions. Our results indicate that parasitic trophic interactions were more accurately identified using compound-specific stable isotope analysis due to removal of underlying source isotopic variation for both parasites and hosts. The compound-specific method provided clearer trophic discrimination factors in comparison to bulk isotope methods. Amino acid compound specific isotope analysis has widely been applied to examine trophic position within food webs, but our analyses suggest that the method is particularly useful for clarifying the feeding strategies for parasitic species. Baseline isotopic information provided by source amino acids allows clear identification of the fractionation from parasite metabolism by integrating underlying isotopic variations from the host tissues. However, like for bulk isotope analysis, the application of a universal trophic discrimination factor to parasite–host relationships remains inappropriate for compound-specific stable isotope analysis. Despite this limitation, compound-specific stable isotope analysis is and will continue to be a valuable tool to increase our understanding of parasitic interactions in marine food webs.

A changing marine environment with emerging natural and anthropogenic stressors challenges the marine mammal immune system. The skin and adnexa form a first protective barrier in the immune response, although this is still relatively understudied in cetaceans. The cellular and tissue morphology of the nodular and diffuse lymphoid tissue are not fully charted and the physiological responses are not yet completely understood. The odontocete’s external ear canal has a complex relationship with the external environment, with an artificial lumen rendering the inside of the canal a relatively secluded environment. In this work, we studied the odontocete ear canal-associated lymphoid tissue (ECALT) by histo- and immunohistochemistry (HC, IHC) with anti-CD3, anti-CD20, anti-Iba-1, anti-HLA-DR, and anti-vimentin antibodies. The ECALT cellular composition consists mainly of B-lymphocytes with the occasional presence of T-lymphocytes and the dispersed distribution of the macrophages. In cases of activation, the cellular reaction showed a similar pattern with the occasional presence of T-cells, plasma cells, and neutrophils. Nodular lymphoid tissue was generally in line with the description in other odontocetes, although with abundant erythrocytes throughout the entire organ. This study contributes to the understanding of the cellular composition of diffuse and nodular lymphoid tissue in several species of odontocetes, and in association with inflammation of the external ear canal.

Long-finned pilot whale (Globicephala melas) mitochondrial (mtDNA) genetic diversity is considered low, especially in the North Atlantic, where only seven haplotypes have been recorded in previous studies using a 345 bp control region fragment. Such studies have not included samples from Ireland or the Netherlands. In this study we analyzed a longer sequence of the mtDNA control region (631 bp) from individuals stranded around Ireland, Scotland, and the Netherlands between 1995 and 2019 (n = 180). Nine haplotypes were identified, of which five were newly described (haplotype diversity h = 0.511). Pairwise tests revealed significant differentiation between the Irish and Scottish samples. Potential confounding factors are discussed but given that failure to recognize population structure may compromise conservation efforts, the findings show the need for further investigation using nuclear markers. Six mass stranding events were included, of which one event reported two haplotypes among individuals confirming a mixing of matrilineal groups. Although the permanence of this combination cannot be determined, this is the first record of such an occurrence within the North Atlantic. This study shows that stranding sample databases are a useful resource for genetic studies and provides new insights into genetic diversity of long-finned pilot whales in the eastern North Atlantic and adjacent waters.

The application of whole-body post-mortem computed tomography (PMCT) in veterinary and wildlife post-mortem research programs is advancing. A high incidence of pulmonary pathology is reported in the harbor porpoise (Phocoena phocoena). In this study, the value of PMCT focused on pulmonary assessment is evaluated. The objectives of this study were to describe pulmonary changes as well as autolytic features detected by PMCT examination and to compare those findings with conventional necropsy. Retrospective evaluation of whole-body PMCT images of 46 relatively fresh harbor porpoises and corresponding conventional necropsy reports was carried out, with a special focus on the respiratory tract. Common pulmonary PMCT findings included: moderate (24/46) to severe (19/46) increased pulmonary soft tissue attenuation, severe parasite burden (17/46), bronchial wall thickening (30/46), and mild autolysis (26/46). Compared to conventional necropsy, PMCT more frequently identified pneumothorax (5/46 vs. none), tracheal content (26/46 vs. 7/46), and macroscopic pulmonary mineralization (23/46 vs. 11/46), and provided more information of the distribution of pulmonary changes. These results indicate that PMCT adds information on pulmonary assessment and is a promising complementary technique for necropsy, despite the frequent presence of mild autolytic features.

Baleen from mysticete whales is a well-preserved proteinaceous material that can be used to identify migrations and feeding habits for species whose migration pathways are unknown. Analysis of δ 13 C and δ 15 N values from bulk baleen have been used to infer migration patterns for individuals. However, this approach has fallen short of identifying migrations between regions as it is difficult to determine variations in isotopic shifts without temporal sampling of prey items. Here, we apply analysis of δ 15 N values of amino acids to five baleen plates belonging to three species, revealing novel insights on trophic position, metabolic state and migration between regions. Humpback and minke whales had higher reconstructed trophic levels than fin whales (3.7–3.8 versus 3–3.2, respectively) as expected due to different feeding specialization. Isotopic niche areas between baleen minima and maxima were well separated, indicating regional resource use for individuals during migration that aligned with isotopic gradients in Atlantic Ocean particulate organic matter. Phenylanine δ 15 N values confirmed regional separation between the niche areas for two fin whales as migrations occurred and elevated glycine and threonine δ 15 N values suggested physiological changes due to fasting. Simultaneous resolution of trophic level and physiological changes allow for identification of regional migrations in mysticetes.

Evidence of hearing impairment was identified in a harbour porpoise (Phocoena phocoena) on the basis of scanning electron microscopy. In addition, based on histopathology and immunohistochemistry, there were signs of unrelated cerebral toxoplasmosis. The six-year old individual live stranded on the Dutch coast at Domburg in 2016 and died a few hours later. The most significant gross lesion was multifocal necrosis and haemorrhage of the cerebrum. Histopathology of the brain revealed extensive necrosis and haemorrhage in the cerebrum with multifocal accumulations of degenerated neutrophils, lymphocytes and macrophages, and perivascular lymphocytic cuffing. The diagnosis of cerebral toxoplasmosis was confirmed by positive staining of protozoa with anti-Toxoplasma gondii antibodies. Tachyzoites were not observed histologically in any of the examined tissues. Ultrastructural evaluation of the inner ear revealed evidence of scattered loss of outer hair cells in a 290 µm long segment of the apical turn of the cochlea, and in a focal region of ~ 1.5 mm from the apex of the cochlea, which was compatible with noise-induced hearing loss. This is the first case of concurrent presumptive noise-induced hearing loss and toxoplasmosis in a free-ranging harbour porpoise from the North Sea.

This report presents a validated list of stranded cetaceans in the Netherlands, as well as some cetaceans accidentally brought in on ship bulbs, between 2015-2019. During this period 2701 cetaceans representing eleven species were reported. The list also includes a few bones and skulls, among others of killer whale (Orcinus orca) and bottlenose dolphin (Tursiops truncatus), species not reported otherwise during this period. The most common species was harbour porpoise (Phocoena phocoena), with 2651 individuals. The average number of porpoises per year was 530, lower than the preceding seven-year average, but numbers fluctuated largely between years and there has been no particular trend after the strong increase in the early 2000s. The monthly pattern of strandings as recorded during the previous periods, with peaks in March and July-September, remained the same. The number of stranded porpoise is equally spread along the entire coastline. The density, expressed as the number of stranded porpoise per kilometre per year, is 0.6 for the entire coastline, or 1.2 if the extensive and less well surveyed areas of Western Scheldt, Eastern Scheldt and Wadden Sea proper are omitted. Sex ratio remained stable over the years and is in line with results from before 2015, with a preponderance of males. On the basis of length, over half were immature and 8.7% neonate. Among neonates and immatures there was a preponderance of males as well, but not in adults. In the Wadden Sea area more neonates and adults were found than in the other two subareas. The major cause of death was infectious disease, followed by predation by grey seal (Halichoerus grypus). Accidental by-catch was identified as the cause of death of 11% of the stranded animals. Of particular interest was the stranding of six sperm whales (Physeter macrocephalus) in January 2016, the largest stranding event of this species in the Netherlands. It was part of a stranding event spread out over the Central and southern North Sea, involving thirty individuals. During 2015-2019 four dead fin whales (Balaenoptera physalus) and eight minke whales (B. acutorostrata) were reported, several of which were hit by ships. There seems to be a slight increase in the strandings of fin whale and Sowerby’s beaked whale (Mesoplodon bidens) since 2000, while white-beaked dolphin (Lagenorhynchus albirostris) shows a decrease. There is no temporal trend for any of the other species since 2000.