title: Accumulation of Emerging Chemicals in Southern Hemisphere Humpback Whales (Megaptera novaeangliae) and Species-specific Chemical Effect Assessment
Creators: name: Casa, Maria V
Publisher: name: Griffith University
Year: 2022
Subjects: subject: southern hemisphere (SH)
subject: humpback whales (Megaptera novaeangliae)
subject: Antarctic
subject: short-chain chlorinated paraffins (SCCPs)
subject: Blubber
subject: induced pluripotent stem cells (iPSCs)
subject: sea-ice ecosystem
subject: consumers

description: Millions of hazardous chemicals are synthesized each year. Their release often represents a risk for the environment and biota. As part of a broader effort to investigate the accumulation and effect of emerging chemicals of concern in the Antarctic sea-ice ecosystem, this thesis focuses upon exposure and effect assessment in southern hemisphere (SH) humpback whales (Megaptera novaeangliae). Cetaceans traditionally represent ideal targets for exposure assessment to Persistent Organic Pollutants (POPs), that is chemicals identified as persistent, bioaccumulative, toxic and capable of long-range environmental transport. Their extended life span and high proportion of body fat render cetaceans susceptible to accumulation of elevated burdens. Southern hemisphere humpback whales feed almost exclusively on Antarctic krill (Euphasia superba) and thus represent second order consumers of the Antarctic sea-ice ecosystem. Exposure assessment consisted of targeted analysis of short-chain chlorinated paraffins (SCCPs) included under the Stockholm Convention (SC) in 2017, and brominated structures listed in 2009 as well as novel, naturally occurring brominated structures, already detected at lower trophic levels of the Antarctic sea-ice ecosystem. SCCPs were targeted in stranded specimens and were detected in 77.7 % of samples, representing the first detection of SCCPs in any mysticetes or any Antarctic foraging marine mammal.Blubber samples from free-swimming and stranded specimens were dedicated to the analysis of the modern category of POPs, polybromodiphenilethers (PBDEs), and a naturally occurring brominated compound, 2,4,6-tribromoanysole (TBA), the chemical structure of which closely resembles PBDEs, flagging the possibility of similar toxic action. Humpback whale PBDEs profiles were dominated by tetra- and penta- congeners, supporting a single previous study on PBDE accumulation in Southern hemisphere humpback whales feeding in the Antarctic Peninsula region. Further, the detection of decaBDE adds support for the local emissions or particle bound transport to Antarctica. PBDE congeners were detected in all samples analysed, whilst TBA was 88 detected in 80% of samples. Upon establishing accumulation of three novel chemical groups in the Antarctic sea-ice ecosystem, species-specific effect assessment was performed through cytotoxicity assessment implementing the newly established immortalized humpback whale fibroblast cell line (HuWaTERT). Two endpoints were targeted, namely membrane integrity and metabolic activity following 24h exposure to an SCCP cocktail, BDE-47, BDE-99, TBA. Each assay also incorporated the assay reference chemical, p,p’-DDE. The four chemicals did not show a dose-dependent response comparable to the one produced by the reference chemical. Whilst p,p’-DDE induced a decrease of both metabolic activity and membrane integrity, the cell response to BDE-99 indicated a positive, or potentially hermetic relationship with regards to the metabolic activity, warranting further investigation of underlying mechanisms.The final thesis chapter explores the unique potential of HuWaTERT for the generation of induced pluripotent stem cells (iPSCs). The main objective of this work was to generate a source of a number of different cell lines from the iPSCs, that could be applied for a more tailored approach to in-vitro chemical effect evaluation to fill the gap currently existing in the field of cetacean toxicology. The re-generation trial involved the use of the Sendai virus, which induces the de-differentiation mechanism in HuWaTERT to become iPSCs without integrating into the cell genome. After successfully generating a number of iPC colonies, the cells ultimately did not thrive, likely due to a fungal infection. Overall, this thesis combines both chemical exposure and effect assessment of novel chemical groups, for unique insight into chemical accumulation and impact in the Antarctic sea-ice ecosystem. The work provides new information regarding the range of chemicals that Antarctic consumers are exposed to, a reflection of hemispheric chemical usage and local input, and also contributes invaluable species-specific in vitro toxicity assessment data, which represents a critical research gap for cetacean chemical impact assessment.