A pregnancy test for whales?! Why and how?

Dr. Alejandro A. Fernández Ajó, Postdoctoral Scholar, Marine Mammal Institute – OSU Department of Fisheries, Wildlife, & Conservation Sciences, Geospatial Ecology of Marine Megafauna (GEMM) Lab.

I often receive two reactions when asked what I am currently working on; one is “Wow! That is a very cool job, it must be amazing to work with such incredible animals!”, the other is “How do you do that and why is that important?”. So, today I decided to blog about some of the reasons why it is important to develop a pregnancy test for gray whales and how we are doing this.

In a previous blogpost, I described the many ways in which whales play critical roles in sustaining marine ecosystem. Briefly, whales can enhance marine productivity by vertically and horizontally mixing of ocean waters, promoting primary production, and mitigating climate change by sequestering carbon with their large biomass and long life-span (1-3). Even after they die, their carcasses can contribute to biodiversity creating new habitat on the seafloor (4). But, over several decades, the whaling industry drastically removed whales around the globe, with some species and populations depleted to near extinction (5). Consequently, these depleted whale populations now play a diminished role in ocean ecosystem processes and their recovery is currently challenged by an increasing number of modern anthropogenic impacts. Hence, working towards whale conservation is essential for keeping a healthy marine ecosystem.

Working and designing effective strategies for conservation biology often involves gaining knowledge regarding the reproductive parameters of individual animals in wild populations. This information is critical for understanding population trends and the underlaying mechanisms that affect animal welfare and their potential for recovery. However, getting such information from free-living whales can be challenging (see Hunt et al. 2013). While we know that whales typically have long life-spans, lengthy generation times, extended parental care, and high survival rates, detailed knowledge on the life history and general reproductive biology of free-ranging whales is limited for the majority of the whale populations. In fact, much of what we do know about whale reproduction is derived from whaling records. Only recently, conservation physiology approaches (see our previous post here) have contributed alternative and non-invasive methods for monitoring key physiological processes that can help monitor a whale’s reproductive biology and determine reproductive parameters such as sexual maturity and pregnancy (6-9).

In this clip you can see an example of a fecal sample collection from a gray whale off the Oregon coast. We can look at hormones in the fecal samples which are useful indicators for endocrine assessments of free-swimming whales. Fecal sample and footage filmed under NOAA/NMFS permit #16111.

Gray whales (Eschrichtius robustus) in the Eastern North Pacific (ENP) typically undertake annual migrations between their lower latitude breeding grounds in the coastal waters of the Baja California Peninsula, Mexico, and the foraging grounds located on the Bering and Chukchi Seas (10). However, among the ENP whales a distinct subgroup of about 230 whales shorten their migration to feed in the coastal waters of Northern California, Oregon, and southeastern Alaska (11). This group of whales is known as the gray whale Pacific Coast Feeding Group (PCFG).

Since 2016, the GEMM Lab has monitored individual gray whales within the PCFG off the Oregon coast (check the GRANITE project). Gray whales have a distinct mottled skin; and each individual whale presents a unique pigmentation pattern that allows for the individual identification of whales. We can identify who is who among the whales who visit the Oregon coast. In this way, we can keep a detailed record of re-sightings of known individuals (visit our new web site to know more about the lives of individual whales that visit the Oregon coast).  We have high individual re-sighting rates, so this unique opportunity helps us keep a long-term data series for individual whales to monitor their health, body condition, and reproductive status over time, and thus further develop and advance our non-invasive study methods.

We are combining behavioral and feeding ecology with drone photogrammetry and endocrinology of the same individual whales to help us understand the relationships between natural and anthropogenic drivers with biological parameters. In this way, following individual whales, we are developing sensitive biomarkers to monitor and infer about the population health, population trends, and identify stressors that impact their recovery and welfare. In particular, we are now working to develop a noninvasive approach to detect pregnancy in gray whales based on fecal hormone analyses.

In this picture you can see “Rose”, a gray whale calf, on top of her mother “Scarlett”. Scarlett is one of the most recognizable whales from the PCFG, due to a large scar on the right side of her back (not visible in this picture). She has been observed along the Pacific NW coast since 1996, so she is at least 26 years old today. We know 3 of her calves. Following individual whales like Scarlett is helping us to better understand the gray whale reproductive biology. Photo by Alejandro Fernandez Ajo taken under NOAA/NMFS permit #21678.

In marine mammals, the progesterone hormone is secreted in the ovaries during the estrous cycle and gestation, and is the predominant hormone responsible for sustaining pregnancy (12). As the hormones are cleared from the blood into the gut, they are metabolized and eventually excreted in feces; fecal samples represent a cumulative and integrated concentration of hormone metabolites (13-14), which are useful indicators for endocrine assessments of free-swimming whales. Several studies show that changes in hormone concentration correlate in meaningful ways with exposure to stressors (15-16) and changes in reproductive status (17-19). We are using our long data series of fecal hormones and individual life histories to advance our understanding on the gray whales’ reproductive biology. We are close to developing a technique that will allow us to detect pregnancy in whales based in fecal hormones analyses and photogrammetry. Stay tuned for results from this pregnancy test!

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1- Pershing AJ, Christensen LB, Record NR, Sherwood GD, Stetson PB (2010) The impact of whaling on the ocean carbon cycle: Why bigger was better. PLoS ONE 5(8): e12444.

2- Roman J and McCarthy JJ. 2010. The whale pump: marine mammals enhance primary productivity in a coastal basin. PLoS ONE. 5(10): e13255.

3- Morissette L, Kaschner K, and Gerber LR. 2010. “Whales eat fish”? Demystifying the myth in the Caribbean marine ecosystem. Fish Fish 11: 388–404.

4- Smith CR, Roman J, Nation JB. A metapopulation model for whale-fall specialists: The largest whales are essential to prevent species extinctions. J. Mar. Res. 77, 283–302 (2019).

5- Branch TA, Williams TM. Legacy of industrial whaling. Whales. Whal. Ocean Ecosyst. 2006, 262–278 (2006).

6- Kellar NM, Keliher J, Trego ML, Catelani KN, Hanns C, George JC, et al. Variation of bowhead whale progesterone concentrations across demographic groups and sample matrices. Endanger Species Res 2013; 22:61–72. https://doi.org/10.3354/esr00537.

7- Pallin L, Robbins J, Kellar N, Berube M, Friedlaender A. Validation of a blubber-based endocrine pregnancy test for humpback whales. Conserv Physiol 2018;6:1 11. https://doi.org/10.1093/conphys/coy031PMID:29942518.

8-Hunt KE, Robbins J, Buck CL, Bérubé M, Rolland RM (2019) Evaluation of fecal hormones for noninvasive research on reproduction and stress in humpback whales (Megaptera novaeangliae). Gen Comp Endocrinol 280: 24–34.

9-Melica, V., Atkinson, S., Calambokidis, J., Lang, A., Scordino, J., & Mueter, F. (2021). Application of endocrine biomarkers to update information on reproductive physiology in gray whale (Eschrichtius robustus). Plos one, 16(8), e0255368.

10-Swartz SL. Gray Whale. In: Wursig B, Thewissen JGM, Kovacs KM, editors. Encyclopedia of Marine Mammals (Third Edition). Elsevier;2018,p. 422–8.https://doi.org/10.1016/B978-0-12-804327-1.00140–0.

11-Calambokidis J, Darling JD, Deecke V, Gearin P, Gosho M, Megill W, et al. Abundance, range and movements of a feeding aggregation of gray whales (Eschrichtius robustus) from California to south-eastern Alaska in 1998. J Cetacean Res Manag 2002;4:267–76.

12- Bronson, F. H. (1989). Mammalian reproductive biology. University of Chicago Press.

13-Wasser SK, Hunt KE, Brown JL, Cooper K, Crockett CM, Bechert U, Millspaugh JJ, Larson S, Monfort SL (2000) A generalized fecal glucocorticoid assay for use in a diverse array of nondomestic mammalian and avian species. Gen Comp Endocrinol120:260–275.

14- Hunt, K.E., Rolland, R.M., Kraus, S.D., Wasser, S.K., 2006. Analysis of fecal glucocorticoids in the North Atlantic right whale (Eubalaena glacialis). Gen. Comp. Endocrinol. 148, 260–272. https://doi.org/10.1016/j.ygcen.2006.03.01215.

15- Lemos, L.S., Olsen, A., Smith, A., Burnett, J.D., Chandler, T.E., Larson, S., Hunt, K.E., Torres, L.G., 2021. Stressed and slim or relaxed and chubby? A simultaneous assessment of gray whale body condition and hormone variability. Mar. Mammal Sci. 1–11. https://doi.org/10.1111/mms.12877

16- Rolland, R., McLellan, W., Moore, M., Harms, C., Burgess, E., Hunt, K., 2017. Fecal glucocorticoids and anthropogenic injury and mortality in North Atlantic right whales Eubalaena glacialis. Endanger. Species Res. 34, 417–429. https://doi.org/10.3354/esr00866.

17-Rolland, R.M., Hunt, K.E., Kraus, S.D., Wasser, S.K., 2005. Assessing reproductive status of right whales (Eubalaena glacialis) using fecal hormone metabolites. Gen. Comp. Endocrinol. 142, 308–317. https://doi.org/10.1016/j.ygcen.2005.02.002

18- Valenzuela Molina M, Atkinson S, Mashburn K, Gendron D, Brownell RL. Fecal steroid hormones reveal reproductive state in female blue whales sampled in the Gulf of California, Mexico. Gen Comp Endocrinol 2018;261:127–35.https://doi.org/10.1016/j.ygcen.2018.02.015 PMID:29476760.

19- Hunt, K. E., Robbins, J., Buck, C. L., Bérubé, M., & Rolland, R. M. (2019). Evaluation of fecal hormones for noninvasive research on reproduction and stress in humpback whales (Megaptera novaeangliae). General and Comparative Endocrinology, 280, 24-34.

Putting Physiological Tools to Work for Whale Conservation

By Alejandro Fernandez Ajo, PhD student at the Department of Biology, Northern Arizona University, Visiting scientist in the GEMM Lab working on the gray whale physiology and ecology project  

About four years ago, I was in Patagonia, Argentina deciding where to focus my research and contribute to whale conservation efforts. At the same time, I was doing fieldwork with the Whale Conservation Institute of Argentina at the “Whale Camp” in Península Valdés. I read tons of papers and talked with my colleagues about different opportunities and gaps in knowledge that I could tackle during my Ph.D. program. One of the questions that caught my attention was about the unknown cause (or causes) for the recurrent high calf mortalities that the Southern Right Whale (SRW) population that breeds at Peninsula Valdés experienced during the 2000s (Rowntree et al. 2013). Still, at that time, I was unsure how to tackle this research question.

Golfo San José, Península Valdés – Argentina. Collecting SRW behavioral data from the cliff’s vantage point. Source: A. Fernandez Ajo.

Between 2003 and 2013, at least 672 SRWs died, of which 91% were calves (Sironi et al. 2014). These mortalities represented an average total whale death per year of 80 individuals in the 2007-2013 period, which vastly exceeded the 8.2 average deaths per year of previous years by a ten-fold increase (i.e., 1993-2002) (Rowntree et al. 2013). In fact, this calf mortality rate was the highest ever documented for any population of large whales. During this period, from 2006 to 2009, I was the Coordinator of the Fauna Area in the Patagonian Coastal Zone Management Plan, and I collaborated with the Southern Right Whale Health Monitoring Program (AKA: The Stranding Program) that conducted field necropsies on stranded whales along the coasts of the Península and collected many different samples including whale baleen.

Southern Right Whale, found stranded in Patagonia Argentina. Source: Instituto de Conservación de Ballenas.

In this process, I learned about the emerging field of Conservation Physiology and the challenges of utilizing traditional approaches to studying physiology in large whales. Basically, the problem is that there is no possible way to obtain blood samples (the gold standard sample type for physiology) from free-swimming whales; whales are just too large! Fortunately, there are currently several alternative approaches for gathering physiological information on large whales using a variety of non-lethal and minimally invasive (or non-invasive) sample matrices, along with utilizing valuable samples recovered at necropsy (Hunt et al. 2013). That is how I learned about Dr. Kathleen Hunt’s novel research studying hormones from whale baleen (Hunt et al., 2018, 2017, 2014). Thus, I contacted Dr. Hunt and started a collaboration to apply these novel methods to understand the case of calf mortalities of the SRW calves in Patagonia utilizing the baleen samples that we recovered with the Stranding program at Península Valdés (see my previous blog post).

What is conservation physiology?

Conservation physiology is a multidisciplinary field of science that utilizes physiological concepts and tools to understand underlying mechanisms of disturbances to solve conservation problems. Conservation physiology approaches can provide sensitive biomarkers of environmental change and allow for targeted conservation strategies. The most common Conservation Physiology applications are monitoring environmental stressors, understanding disease dynamics and reproductive biology, and ultimately reducing human-wildlife conflict, among other applications.

I am now completing the last semester of my Ph.D. program. I have learned much about the amazing field of Conservation Physiology and how much more we need to know to achieve our conservation goals. I am still learning, yet I feel that through my research I have contributed to understanding how different stressors impact the health and wellbeing of whales, and about aspects of their biology that have long been obscured or unknown for these giants. One contribution I am proud of is our recent publication of, “A tale of two whales: putting physiological tools to work for North Atlantic and southern right whales,” which was published in January 2021 as a book chapter in “Conservation Physiology: Applications for Wildlife Conservation and Management” published by Oxford University Press: Oxford, UK.

This book outlines the significant avenues and advances that conservation physiology contributes to the monitoring, management, and restoration of wild animal populations. The book also defines opportunities for further growth in the field and identifies critical areas for future investigation. The text and the contributed chapters illustrate several examples of the different approaches that the conservation physiology toolbox can tackle. In our chapter, “A tale of two whales,” we discuss developments in conservation physiology research of large whales, with the focus on the North Atlantic right whale (Eubalaena glacialis) and southern right whale (Eubalaena australis), two closely related species that differ vastly in population status and conservation pressures. We review the advances in Conservation Physiology that help overcome the challenges of studying large whales via a suite of creative approaches, including photo-identification, visual health assessment, remote methods of assessing body condition, and endocrine research using non-plasma sample types such as feces, respiratory vapor, and baleen. These efforts have illuminated conservation-relevant physiological questions for both species, such as discrimination of acute from chronic stress, identification of likely causes of mortality, and monitoring causes and consequences of body condition and reproduction changes.

Book Overview:

This book provides an overview of the different applications of Conservation Physiology, outlining the significant avenues and advances by which conservation physiology contributes to the monitoring, management, and restoration of wild animal populations. By using a series of global case studies, contributors illustrate how approaches from the conservation physiology toolbox can tackle a diverse range of conservation issues, including monitoring environmental stress, predicting the impact of climate change, understanding disease dynamics, and improving captive breeding, and reducing human-wildlife conflict. The variety of taxa, biological scales, and ecosystems is highlighted to illustrate the far-reaching nature of the discipline and allow readers to appreciate the purpose, value, applicability, and status of the field of conservation physiology. This book is an accessible supplementary textbook suitable for graduate students, researchers, and practitioners in conservation science, ecophysiology, evolutionary and comparative physiology, natural resources management, ecosystem health, veterinary medicine, animal physiology, and ecology.


Hunt KE, Fernández Ajó A, Lowe C, Burgess EA, Buck CL. 2021. A tale of two whales: putting physiological tools to work for North Atlantic and southern right whales. In: “Conservation Physiology: Integrating Physiology Into Animal Conservation And Management”, ch. 12. Eds. Madliger CL, Franklin CE, Love OP, Cooke SJ. Oxford University press: Oxford, UK.

Sironi, M., Rowntree, V., Di Martino, M. D., Beltramino, L., Rago, V., Franco, M., and Uhart, M. (2014). Updated information for 2012-2013 on southern right whale mortalities at Península Valdés, Argentina. SC/65b/BRG/06 report presented to the International Whaling Commission Scientific Committee, Portugal. <https://iwc.int/home>.

Rowntree, V.J., Uhart, M.M., Sironi, M., Chirife, A., Di Martino, M., La Sala, L., Musmeci, L., Mohamed, N., Andrejuk, J., McAloose, D., Sala, J., Carribero, A., Rally, H., Franco, M., Adler, F., Brownell, R. Jr, Seger, J., Rowles, T., 2013. Unexplained recurring high mortality of southern right whale Eubalaena australis calves at Península Valdés, Argentina. Marine Ecology Progress Series, 493, 275-289. DOI: 10.3354/meps10506

Hunt KE, Moore MJ, Rolland RM, Kellar NM, Hall AJ, Kershaw J, Raverty SA, Davis CE, Yeates LC, Fauquier DA, et al., 2013. Overcoming the challenges of studying conservation physiology in large whales: a review of available methods. Conserv Physiol 1: cot006–cot006.

Hunt, K.E., Stimmelmayr, R., George, C., Hanns, C., Suydam, R., Brower, H., Rolland, R.M., 2014. Baleen hormones: a novel tool for retrospective assessment of stress and reproduction in bowhead whales (Balaena mysticetus). Conserv. Physiol. 2, cou030. https://doi.org/10.1093/conphys/cou030

Hunt, K.E., Lysiak, N.S., Moore, M.J., Rolland, R.M., 2016. Longitudinal progesterone profiles in baleen from female North Atlantic right whales (Eubalaena glacialis) match known calving history. Conserv. Physiol. 4, cow014. https://doi.org/10.1093/conphys/cow014

Hunt, K.E., Lysiak, N.S., Robbins, J., Moore, M.J., Seton, R.E., Torres, L., Buck, C.L., 2017. Multiple steroid and thyroid hormones detected in baleen from eight whale species. Conserv. Physiol. 5. https://doi.org/10.1093/conphys/cox061

Hunt, K.E., Lysiak, N.S.J., Matthews, C.J.D., Lowe, C., Fernández Ajó, A., Dillon, D., Willing, C., Heide-Jørgensen, M.P., Ferguson, S.H., Moore, M.J., Buck, C.L., 2018. Multi-year patterns in testosterone, cortisol and corticosterone in baleen from adult males of three whale species. Conserv. Physiol. 6, coy049. https://doi.org/10.1093/conphys/coy049