A non-invasive approach to pregnancy diagnosis in Gray whales is possible!

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

In a previous post (link to blog), I discussed the crucial importance of acquiring knowledge on the reproductive parameters of individual animals in wild populations for designing effective strategies in conservation biology. Specifically, the ability to quantify the number of pregnancies within a population offers valuable insights into the health of individual females and the population as a whole [1,2]. This knowledge provides tools to describe important life-history parameters, including the age of sexual maturity, frequency of pregnancy, duration of gestation, timing of reproduction, and population fecundity; all of which are essential components for monitoring trends in reproduction and the overall health of a species [3]. Additionally, I explained some of the challenges inherent in obtaining such information when working with massive wild animals that spend most of their time underwater in vast expanses of the oceans. Yes, I am talking about whales.

As a result of the logistical and methodological challenges that involve the study of large whales, detailed knowledge of the life-history and general reproductive biology of whales is sparse for most species and populations. In fact, much of the available information is derived from whaling records [4], which may be outdated for application in population models [5].

If you are an avid reader of the GEMM Lab blog posts, you might be familiar with the gray whale (Eschrichtius robustus), and with the distinct subgroup of gray whales, known as the Pacific Coast Feeding Group (PCFG). PCFG gray whales are characterized by their shorter migration to spend their feeding season in the coastal waters of Northern California, Oregon, and southeastern Alaska [6], relative to the larger Eastern North Pacific gray whale that forage in the Arctic region.

The GEMM Lab has monitored individual gray whales within the PCFG off the Oregon coast since 2016 (check the GRANITE project). Each individual whale presents a unique pigmentation pattern, or unique marks that we can use to identify who is who among the whales who visit the Oregon coast. In this way, we keep a detailed record of re-sightings of known individuals (visit IndividuWhale to learn more), and we have high individual re-sighting rates, resulting in a long-term data series for individual whales which enables us to monitor their health, body condition, and thus further develop and advance our non-invasive study methods.

Drone-based image of a Gray whale defecating. Source: GEMM Lab, NOAA/NSF permit #16111

In our recently manuscript published in the Royal Society Open Science journal, armed with our robust dataset comprising fecal hormone metabolites, drone-based photogrammetry, and individual sightings, we delved into the strengths and weaknesses of various diagnostic tools for non-invasive pregnancy diagnosis. Ultimately, we propose a methodological approach that can help with the challenging and important task of identifying pregnancies in gray whales. In particular, we explored the variability in fecal progesterone metabolites and body morphology relative to observed reproductive status and estimated the pregnancy probability for mature females using statistical models.

In mammals, the progesterone hormone is secreted in the ovaries during the estrous cycle and gestation, making it the predominant hormone responsible for sustaining pregnancy [7]. 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 [8;9], which are useful indicators for endocrine assessments of free-swimming whales. Additionally, our previous studies in this population [10] detected differences in body condition (see KC blog for more details about how we measure whales) that suggest that changes in the whale’s body widths could be useful in detecting pregnancies.

Our exploratory analyses show that in individual whales, the levels of fecal progesterone were elevated when pregnant as compared to when the same whale was not pregnant. But when looking at progesterone levels at the population level, these differences were masked with the intrinsic variability of this measurement. In turn, the body morphometrics, in particular the body width at the 50% of the total body length, helped discriminate pregnancies better, and the statistical models that included this width variable, effectively classified pregnant from non-pregnant females with a commendable accuracy. Thus, our morphometric approach showcased its potential as a reliable alternative for pregnancy diagnosis.

Below, a comparison of body widths at 5% increments along total body length (from 20 % to 70 %) in female gray whales of known reproductive status from UAS-based photogrammetry (example photograph shown at top). Pregnant females (PF; in blue), presumed nonpregnant juvenile females (JF; yellow), and lactating females (LF; orange). Fernandez Ajó et al. 2023.

Notably, when we ran the pregnancy prediction models on data from our 2022 season and compared results with observations of whales in 2023, we identified a known whale from our study area “Clouds” accompanied by a calf, indicating that she was pregnant in 2022. Our model predicted Clouds to be pregnant with a 70% probability. This validation lends strong confidence to our approach to diagnosing pregnancy. Conversely, some whales predicted to be pregnant in 2022 were not observed with a calf during the 2023 season. However, the absence of calves accompanying these females is likely due to the relatively high mortality of newborn calves in gray whales due to predation or other causes [11].

Overall, our findings underscore some limitations of fecal progesterone metabolite in accurately identifying pregnant PCFG gray whales. However, while acknowledging the challenges associated with fecal sample collection and hormone analysis, we advocate for ongoing exploration of alternative hormone quantification methods and antibodies. Our study highlights the importance of continued research in refining these techniques. The unique attributes of our study system, including high individual re-sighting rates and non-invasive fecal hormone analysis, position it as a cornerstone for future advancements in understanding gray whale reproductive health. By improving our ability to monitor reproductive metrics in baleen whale populations, we pave the way for more effective conservation strategies, ensuring the resilience of these magnificent creatures in the face of a changing marine ecosystems.



[1] Burgess EA, Lanyon JM, Brown JL, Blyde D, Keeley T. 2012 Diagnosing pregnancy in free-ranging dugongs using fecal progesterone metabolite concentrations and body morphometrics: A population application. Gen Comp Endocrinol 177, 82–92. (doi:10.1016/J.YGCEN.2012.02.008)

[2] Slade NA, Tuljapurkar S, Caswell H. 1998 Structured-Population Models in Marine, Terrestrial, and Freshwater Systems. J Wildl Manage 62. (doi:10.2307/3802363)

[3] Madliger CL, Love OP, Hultine KR, Cooke SJ. 2018 The conservation physiology toolbox: status and opportunities. Conserv Physiol 6, 1–16. (doi:10.1093/conphys/coy029)

[4] Rice DW, Wolman AA. 1971 Life history and ecology of the gray whale (Eschrichtius robustus). Stillwater, Oklahoma: American Society of Mammalogists.

[5] Melicai 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. (doi:10.1371/journal.pone.0255368)

[6] 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.

[7] Bronson, F. H. (1989). Mammalian reproductive biology. University of Chicago Press.

[8] 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.

[9] 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.

[10] Soledade Lemos L, Burnett JD, Chandler TE, Sumich JL, Torres LG. 2020 Intra‐ and inter‐annual variation in gray whale body condition on a foraging ground. Ecosphere 11. (doi:10.1002/ecs2.3094)

[11] James L. Sumich, James T. Harvey, Juvenile Mortality in Gray Whales (Eschrichtius robustus), Journal of Mammalogy, Volume 67, Issue 1, 25 February 1986, Pages 179–182, https://doi.org/10.2307/1381019

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