RESUMO
Baleen whales are subject to a myriad of natural and anthropogenic stressors, but understanding how these stressors affect physiology is difficult. Measurement of adrenal glucocorticoid (GC) hormones involved in the vertebrate stress response (cortisol and corticosterone) in baleen could help fill this data gap. Baleen analysis is a powerful tool, allowing for a retrospective re-creation of multiple years of GC hormone concentrations at approximately a monthly resolution. We hypothesized that whales that died from acute causes (e.g. ship strike) would have lower levels of baleen GCs than whales that died from extended illness or injury (e.g. long-term entanglement in fishing gear). To test this hypothesis, we extracted hormones from baleen plates of four humpback whales (Megaptera novaeangliae) with well-documented deaths including multiple and chronic entanglements (n = 1, female), ship strike (n = 2, male and female) and chronic illness with nutritional stress (n = 1, male). Over ~3 years of baleen growth and during multiple entanglements, the entangled whale had average corticosterone levels of 80-187% higher than the other three whales but cortisol levels were similar to two of the other three whales. The nutritionally stressed and chronically ill whale showed a slow increase in both cortisol and corticosterone spanning ~3 years, followed by a sharp decline in both hormones before death, possibly indicative of adrenal failure in this moribund individual. This whale's correlation between cortisol and corticosterone was significant but there were no correlations in the other three whales. Our results show that cortisol and corticosterone concentrations vary according to the type and duration of illness or injury. Single-point GC concentrations should be interpreted with caution as low values can occur in whales experiencing pronounced stress and individual baselines can be highly variable. Baleen analysis is a promising tissue type for retrospective analyses of physiological responses to various stressors affecting baleen whales.
RESUMO
Recent studies have demonstrated that some hormones are present in baleen powder from bowhead (Balaena mysticetus) and North Atlantic right (Eubalaena glacialis) whales. To test the potential generalizability of this technique for studies of stress and reproduction in large whales, we sought to determine whether all major classes of steroid and thyroid hormones are detectable in baleen, and whether these hormones are detectable in other mysticetes. Powdered baleen samples were recovered from single specimens of North Atlantic right, bowhead, blue (Balaenoptera [B.]musculus), sei (B. borealis), minke (B. acutorostrata), fin (B. physalus), humpback (Megaptera novaeangliae) and gray (Eschrichtius robustus) whales. Hormones were extracted with a methanol vortex method, after which we tested all species with commercial enzyme immunoassays (EIAs, Arbor Assays) for progesterone, testosterone, 17ß-estradiol, cortisol, corticosterone, aldosterone, thyroxine and tri-iodothyronine, representing a wide array of steroid and thyroid hormones of interest for whale physiology research. In total, 64 parallelism tests (8 species × 8 hormones) were evaluated to verify good binding affinity of the assay antibodies to hormones in baleen. We also tested assay accuracy, although available sample volume limited this test to progesterone, testosterone and cortisol. All tested hormones were detectable in baleen powder of all species, and all assays passed parallelism and accuracy tests. Although only single individuals were tested, the consistent detectability of all hormones in all species indicates that baleen hormone analysis is likely applicable to a broad range of mysticetes, and that the EIA kits tested here perform well with baleen extract. Quantification of hormones in baleen may be a suitable technique with which to explore questions that have historically been difficult to address in large whales, including pregnancy and inter-calving interval, age of sexual maturation, timing and duration of seasonal reproductive cycles, adrenal physiology and metabolic rate.
