Recent advances in analytical techniques have allowed cetacean pregnancy status to be diagnosed using reproductive hormones from biopsy samples. We tested the efficacy of blubber progesterone assays for diagnosing pregnancy in humpback whales, calculated pregnancy rates and examined the relationship between pregnancy and stable isotope ratios for these whales.

Following the Deepwater Horizon (DWH) oil spill, reproductive success rates in 2 northern Gulf of Mexico (GoM) bottlenose dolphin stocks exposed to oil were evaluated for 4 yr during and after the spill (2010 to 2015) in efforts to assess population-level reproductive health. Pregnancy was determined from either (1) ultrasound examinations of the reproductive tract during capture-release health assessments, or (2) endocrine evaluations of blubber tissue collected from dart biopsies of free-ranging dolphins. Follow-up photo-identification was then used to track the status of pregnant females and any associated neonatal calves for a minimum of 1 yr after the initial pregnancy detection (IPD). For all pregnant females observed following IPD, individuals seen with a calf (reproductive success) and without one (reproductive failure) were recorded. The resulting estimated reproductive success rates for both GoM stocks (19.4%; 7/36) were less than a third of those previously reported in other areas not impacted by the spill (i.e. Sarasota Bay, FL; Indian River Lagoon, FL; and Charleston Harbor, SC) using similar techniques (64.7%; 22/34). We also evaluated the relationships between reproductive success and 13 potential covariates, including stock, ordinal date, progesterone, cortisol, thyroid hormone concentrations, leukocyte count, lung health score, and total body length. Among these, the results only provide strong evidence (Bayes factor > 20) of a relationship between reproductive failure and the total leukocyte count covariate. The high reproductive failure rates measured in both GoM stocks following the DWH oil spill are consistent with mammalian literature that shows a link between petroleum exposure and reproductive abnormalities and failures.

When paired with dart biopsying, quantifying cortisol in blubber tissue may provide an index of relative stress levels (i.e., activation of the hypothalamus-pituitary-adrenal axis) in free-ranging cetacean populations while minimizing the effects of the act of sampling. To validate this approach, cortisol was extracted from blubber samples collected from beach-stranded and bycaught short-beaked common dolphins using a modified blubber steroid isolation technique and measured via commercially available enzyme immunoassays. The measurements exhibited appropriate quality characteristics when analyzed via a bootstraped stepwise parallelism analysis (observed/expected = 1.03, 95%CI: 99.6 – 1.08) and showed no evidence of matrix interference with increasing sample size across typical biopsy tissue masses (75–150mg; r2 = 0.012, p = 0.78, slope = 0.022ngcortisol deviation/ultissue extract added). The relationships between blubber cortisol and eight potential cofactors namely, 1) fatality type (e.g., stranded or bycaught), 2) specimen condition (state of decomposition), 3) total body length, 4) sex, 5) sexual maturity state, 6) pregnancy status, 7) lactation state, and 8) adrenal mass, were assessed using a Bayesian generalized linear model averaging technique. Fatality type was the only factor correlated with blubber cortisol, and the magnitude of the effect size was substantial: beach-stranded individuals had on average 6.1-fold higher cortisol levels than those of bycaught individuals. Because of the difference in conditions surrounding these two fatality types, we interpret this relationship as evidence that blubber cortisol is indicative of stress response. We found no evidence of seasonal variation or a relationship between cortisol and the remaining cofactors.