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Extraction of chemicals from biota leads to co-extraction of lipids. When dosing such extracts into in vitro bioassays, co-dosed lipids act as an additional phase that can reduce the bioavailability of the chemicals and the apparent sensitivity of the assay. Equilibrium partitioning between medium, cells, and co-dosed lipids was described with an existing equilibrium partitioning model for cell-based bioassays extended by an additional lipid phase. We experimentally investigated the influence of co-dosed lipids on the effects elicited by four test chemicals of different hydrophobicity in two bioassays, indicative of the aryl hydrocarbon receptor and oxidative stress response (AREc32). The partitioning model explained the effect of the test chemicals in the presence of spiked triolein within a factor of 0.33−5.83 between the measured and predicted effect concentration (EC). We applied the model to marine mammal blubber extracted with silicone. Extracts dosed in the AREc32 bioassay showed a linear increase of apparent EC with increasing lipid fraction. The partitioning model was used to interpret the role of the co-extracted lipid. A quantitative lipid correction of bioassay results in the presence of co-dosed lipids was possible for known compounds and defined mixtures, while we could only estimate a range for mixtures of unknown chemicals.

The analysis of mixtures of environmental contaminants from marine mammal organs revealed distinct distribution patterns for single compounds. The combination of chemical analysis and bioassays can comprehensively characterize the mixture exposome.

Detailed post-mortem investigations including the auditory pathway are needed to advance our understanding of how underwater noise and other stressors affect hearing in cetaceans. A 12-year-old female porpoise (Phocoena phocoena) stranded alive in June 2021 at the German Baltic Sea coast and died some hours later. The most significant pathological findings were lesions caused by a severe aspergillosis that spread from the lung and pulmonary lymph node to the cerebellum. Based on molecular sequencing, the fungus was identified as Aspergillus fumigatus. Severe pyogranulomatous and necrotizing inflammation was diagnosed in the lung and the associated lymph node. In the left part of the cerebellum, focal, severe purulent and necrotizing meningoencephalitis with intralesional fungal structures was confirmed histologically. In addition, multifocal, severe, chronic, granulomatous, and eosinophilic gastritis with intralesional parasite structures was found in the stomach. Parallel stripes (linear skin markings) were detected along the caudal part of both body sides, which have not been previously described for harbor porpoises. Inner ear analysis revealed evidence of focal loss of outer hair cells in several regions from 120 to 580 µm from the apex of the right cochlea using immunofluorescence. The evidence of low-frequency hearing impairment was compatible with noise-induced hearing loss. This is the first case of concurrent presumptive noise-induced hearing loss and unrelated aspergillosis in a free-ranging harbor porpoise.

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