Hunt
Kathleen E.
Hunt
Kathleen E.
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ArticleFecal glucocorticoids and anthropogenic injury and mortality in North Atlantic right whales Eubalaena glacialis(Inter-Research, 2017-11-30) Rolland, Rosalind M. ; McLellan, William A. ; Moore, Michael J. ; Harms, Craig A. ; Burgess, Elizabeth A. ; Hunt, Kathleen E.As human impacts on marine ecosystems escalate, there is increasing interest in quantifying sub-lethal physiological and pathological responses of marine mammals. Glucocorticoid hormones are commonly used to assess stress responses to anthropogenic factors in wildlife. While obtaining blood samples to measure circulating hormones is not currently feasible for free-swimming large whales, immunoassay of fecal glucocorticoid metabolites (fGCs) has been validated for North Atlantic right whales Eubalaena glacialis (NARW). Using a general linear model, we compared fGC concentrations in right whales chronically entangled in fishing gear (n = 6) or live-stranded (n = 1), with right whales quickly killed by vessels (n = 5) and healthy right whales (n = 113) to characterize fGC responses to acute vs. chronic stressors. fGCs in entangled whales (mean ± SE: 1856.4 ± 1644.9 ng g-1) and the stranded whale (5740.7 ng g-1) were significantly higher than in whales killed by vessels (46.2 ± 19.2 ng g-1) and healthy whales (51.7 ± 8.7 ng g-1). Paired feces and serum collected from the live-stranded right whale provided comparison of fGCs in 2 matrices in a chronically stressed whale. Serum cortisol and corticosterone in this whale (50.0 and 29.0 ng ml-1, respectively) were much higher than values reported in other cetaceans, in concordance with extremely elevated fGCs. Meaningful patterns in fGC concentration related to acute vs. chronic impacts persisted despite potential for bacterial degradation of hormone metabolites in dead whales. These results provide biological validation for using fGCs as a biomarker of chronic stress in NARWs.
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ArticleLongitudinal progesterone profiles in baleen from female North Atlantic right whales (Eubalaena glacialis) match known calving history(Oxford University Press, 2016-03) Hunt, Kathleen E. ; Lysiak, Nadine S. J. ; Moore, Michael J. ; Rolland, Rosalind M.Reproduction of mysticete whales is difficult to monitor, and basic parameters, such as pregnancy rate and inter-calving interval, remain unknown for many populations. We hypothesized that baleen plates (keratinous strips that grow downward from the palate of mysticete whales) might record previous pregnancies, in the form of high-progesterone regions in the sections of baleen that grew while the whale was pregnant. To test this hypothesis, longitudinal baleen progesterone profiles from two adult female North Atlantic right whales (Eubalaena glacialis) that died as a result of ship strike were compared with dates of known pregnancies inferred from calf sightings and post-mortem data. We sampled a full-length baleen plate from each female at 4 cm intervals from base (newest baleen) to tip (oldest baleen), each interval representing ∼60 days of baleen growth, with high-progesterone areas then sampled at 2 or 1 cm intervals. Pulverized baleen powder was assayed for progesterone using enzyme immunoassay. The date of growth of each sampling location on the baleen plate was estimated based on the distance from the base of the plate and baleen growth rates derived from annual cycles of stable isotope ratios. Baleen progesterone profiles from both whales showed dramatic elevations (two orders of magnitude higher than baseline) in areas corresponding to known pregnancies. Baleen hormone analysis shows great potential for estimation of recent reproductive history, inter-calving interval and general reproductive biology in this species and, possibly, in other mysticete whales.
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ArticleMultiple steroid and thyroid hormones detected in baleen from eight whale species(Oxford University Press, 2017-11-09) Hunt, Kathleen E. ; Lysiak, Nadine S. J. ; Robbins, Jooke ; Moore, Michael J. ; Seton, Rosemary E. ; Torres, Leigh ; Buck, C. LorenRecent 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, 17β-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.
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ArticleMulti-year patterns in testosterone, cortisol and corticosterone in baleen from adult males of three whale species(Oxford University Press, 2018-09-21) Hunt, Kathleen E. ; Lysiak, Nadine S. J. ; Matthews, Cory J. D. ; Lowe, Carley ; Ajo, Alejandro Fernández ; Dillon, Danielle ; Willing, Cornelia ; Heide-Jørgensen, Mads Peter ; Ferguson, Steven H. ; Moore, Michael J. ; Buck, C. LorenMale baleen whales have long been suspected to have annual cycles in testosterone, but due to difficulty in collecting endocrine samples, little direct evidence exists to confirm this hypothesis. Potential influences of stress or adrenal stress hormones (cortisol, corticosterone) on male reproduction have also been difficult to study. Baleen has recently been shown to accumulate steroid hormones during growth, such that a single baleen plate contains a continuous, multi-year retrospective record of the whale’s endocrine history. As a preliminary investigation into potential testosterone cyclicity in male whales and influences of stress, we determined patterns in immunoreactive testosterone, two glucocorticoids (cortisol and corticosterone), and stable-isotope (SI) ratios, across the full length of baleen plates from a bowhead whale (Balaena mysticetus), a North Atlantic right whale (Eubalaena glacialis) and a blue whale (Balaenoptera musculus), all adult males. Baleen was subsampled at 2 cm (bowhead, right) or 1 cm (blue) intervals and hormones were extracted from baleen powder with methanol, followed by quantification of all three hormones using enzyme immunoassays validated for baleen extract of these species. Baleen of all three males contained regularly spaced peaks in testosterone content, with number and spacing of testosterone peaks corresponding well to SI data and to species-specific estimates of annual baleen growth rate. Cortisol and corticosterone exhibited some peaks that co-occurred with testosterone peaks, while other glucocorticoid peaks occurred independent of testosterone peaks. The right whale had unusually high glucocorticoids during a period with a known entanglement in fishing gear and a possible disease episode; in the subsequent year, testosterone was unusually low. Further study of baleen testosterone patterns in male whales could help clarify conservation- and management-related questions such as age of sexual maturity, location and season of breeding, and the potential effect of anthropogenic and natural stressors on male testosterone cycles.
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PreprintMulti-year longitudinal profiles of cortisol and corticosterone recovered from baleen of North Atlantic right whales (Eubalaena glacialis)( 2017-09) Hunt, Kathleen E. ; Lysiak, Nadine S. J. ; Moore, Michael J. ; Rolland, Rosalind M.Research into stress physiology of mysticete whales has been hampered by difficulty in obtaining repeated physiological samples from individuals over time. We investigated whether multi-year longitudinal records of glucocorticoids can be reconstructed from serial sampling along full-length baleen plates (representing ~10 years of baleen growth), using baleen recovered from two female North Atlantic right whales (Eubalaena glacialis) of known reproductive history. Cortisol and corticosterone were quantified with immunoassay of subsamples taken every 4 cm (representing ~60 d time intervals) along a full-length baleen plate from each female. In both whales, corticosterone was significantly elevated during known pregnancies (inferred from calf sightings and necropsy data) as compared to intercalving intervals; cortisol was significantly elevated during pregnancies in one female but not the other. Within intercalving intervals, corticosterone was significantly elevated during the first year (lactation year) and/or the second year (post-lactation year) as compared to later years of the intercalving interval, while cortisol showed more variable patterns. Cortisol occasionally showed brief high elevations (“spikes”) not paralleled by corticosterone, suggesting that the two glucocorticoids might be differentially responsive to certain stressors. Generally, immunoreactive corticosterone was present in higher concentration in baleen than immunoreactive cortisol; corticosterone:cortisol ratio was usually >4 and was highly variable in both individuals. Further investigation of baleen cortisol and corticosterone profiles could prove fruitful for elucidating long-term, multi-year patterns in stress physiology of large whales, determined retrospectively from stranded or archived specimens.
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ArticleOvercoming the challenges of studying conservation physiology in large whales : a review of available methods(Oxford University Press, 2013-05-15) Hunt, Kathleen E. ; Moore, Michael J. ; Rolland, Rosalind M. ; Kellar, Nicholas M. ; Hall, Ailsa J. ; Kershaw, Joanna ; Raverty, Stephen A. ; Davis, Cristina E. ; Yeates, Laura C. ; Fauquier, Deborah A. ; Rowles, Teresa K. ; Kraus, Scott D.Large whales are subjected to a variety of conservation pressures that could be better monitored and managed if physiological information could be gathered readily from free-swimming whales. However, traditional approaches to studying physiology have been impractical for large whales, because there is no routine method for capture of the largest species and there is presently no practical method of obtaining blood samples from free-swimming whales. We review the currently available techniques for gathering physiological information on large whales using a variety of non-lethal and minimally invasive (or non-invasive) sample matrices. We focus on methods that should produce information relevant to conservation physiology, e.g. measures relevant to stress physiology, reproductive status, nutritional status, immune response, health, and disease. The following four types of samples are discussed: faecal samples, respiratory samples (‘blow’), skin/blubber samples, and photographs. Faecal samples have historically been used for diet analysis but increasingly are also used for hormonal analyses, as well as for assessment of exposure to toxins, pollutants, and parasites. Blow samples contain many hormones as well as respiratory microbes, a diverse array of metabolites, and a variety of immune-related substances. Biopsy dart samples are widely used for genetic, contaminant, and fatty-acid analyses and are now being used for endocrine studies along with proteomic and transcriptomic approaches. Photographic analyses have benefited from recently developed quantitative techniques allowing assessment of skin condition, ectoparasite load, and nutritional status, along with wounds and scars from ship strikes and fishing gear entanglement. Field application of these techniques has the potential to improve our understanding of the physiology of large whales greatly, better enabling assessment of the relative impacts of many anthropogenic and ecological pressures.