Validation of a blubber-based endocrine pregnancy test for humpback whales

Baleen whales have few identifiable external indicators of pregnancy state, making it challenging to study essential aspects of their biology and population dynamics. Pregnancy status in other marine mammals has been determined by measuring progesterone concentrations from a variety of sample matrices, but logistical constraints have limited such studies in free-swimming baleen whales. We use an extensive blubber sample archive and associated calving history data to retrospectively identify samples that correspond to pregnant females and develop a progesterone-based pregnancy test for humpback whales. The lowest pregnant blubber progesterone concentration was 54.97 ng g−1, and the mean for the known-pregnant group was 198.74 ± 180.65 ng g−1. Conversely, females known to be below the minimum age of sexual maturity (juvenile females) had an overall low mean progesterone concentration (0.59 ± 0.25 ng g−1), well below the known-pregnant range. Of the mature females that did not return with a calf (n = 11), three fell within the known-pregnant range (320.79 ± 209.34 ng g−1), while the levels for the remaining eight were two orders of magnitude below the lowest known-pregnant level (1.63 ± 1.15 ng g−1). The proportion of females that did not return with a calf but had values similar to known-pregnant females are consistent with rates of calf mortality, but other potential explanations were considered. Our findings support a validated blubber endocrine assignment of pregnancy corroborated with field life history information, a first for any baleen whale species. The progesterone values we measured were similar to those found in different pregnancy states of other cetaceans and support using blubber biopsy samples for assigning pregnancy in humpback whales. This method can be applied to existing archives or new samples to better study life history and population demography broadly across species and populations

Demographic and physiological signals of reproductive events in humpback whales on a southwest pacific breeding ground

The field of marine mammal conservation has dramatically benefited from the rapid advancement of methods to assess the reproductive physiology of individuals and populations from steroid hormones isolated from minimally invasive skin-blubber biopsy samples. Historically, this vital information was only available from complete anatomical and physiological investigations of samples collected during commercial or indigenous whaling. Humpback whales (Megaptera novaeangliae) are a migratory, cosmopolitan species that reproduce in warm, low-latitude breeding grounds. New Caledonia is seasonally visited by a small breeding sub-stock of humpback whales, forming part of the endangered Oceania subpopulation. To better understand the demographic and seasonal patterns of reproductive physiology in humpback whales, we quantified baseline measurements of reproductive hormones (progesterone – P4, testosterone - T, and 17β-estradiol – E2) using an extensive archive of skin-blubber biopsy samples collected from female humpback whales in New Caledonia waters between 2016-2019 (n = 194). We observed significant differences in the P4, T, and E2 concentrations across different demographic groups of female humpback whales, and we described some of the first evidence of the endocrine patterns of estrus in live free-ranging baleen whales. This study is fundamental in its methodological approach to a wild species that has a global distribution, with seasonally distinct life histories. This information will assist in monitoring, managing, and conserving this population as global ecological changes continue to occur unhindered.Peer reviewe

Using Tissue Biomarkers to Understand the Demography and Recovery of Baleen Whales in a Rapidly Changing Environment

Baleen whale populations in the Southern Ocean are recovering after intense commercial whaling in the 20th century. Along the Western Antarctic Peninsula (WAP), this recovery is occurring in one of the planet's most rapidly changing marine ecosystems. Understanding how climate-driven changes influence the population dynamics of whales in this region is critical for understanding what conservation and management actions must be prioritized to maintain the structure and function of this marine ecosystem. This is even more important as this region has seen extraordinary increases in industrial krill fishery pressure, which overlaps in both time and space with whales foraging in this region, as well as increased human presence in the form of ecotourism. Thus, to begin understanding the dynamics of whale recovery under continued environmental change, we need to study these whales' demography and population dynamics. My dissertation aimed to examine and describe the demographics and population dynamics of two species of Southern Hemisphere baleen whales (Antarctic minke whales and Southern Hemisphere humpback whales) in the context of a rapidly changing ecosystem. To do this, I used one of the most extensive, non-lethal tissue archives of these two species, collected as part of the National Science Foundations (NSF) Palmer Station long-term ecological research (LTER) project. I found that on average, Antarctic minke whales reproduced each year and estimated that two-thirds of females along the WAP were sexually mature. More importantly, these data represent the first non-lethal approach to studying this species. Furthermore, I found that broad-scale environmental variation affecting krill abundance and availability along the WAP adversely impacted humpback whale pregnancy rates. This indicates that continued warming along the WAP that results in subsequent changes in the distribution and abundance of prey may adversely affect the recovery of this humpback whale population. Lastly, I found that blubber cortisol levels were not significantly different between male and female humpback whales but were significantly different across different demographic groups of females and across months. Blubber cortisol levels also significantly decreased in 2021, a year when human presence along the WAP was greatly reduced due to the COVID-19 pandemic. These findings provide a critical baseline of cortisol levels for whales in a rapidly changing region and show direct relationships between cortisol levels and human presence. These are some of the first non-lethal quantitative observations of the demography and population dynamics of recovering whale populations in the Antarctic and provide a critical reference point for future work as the Antarctic climate continues to change and populations continue to recover from whaling.

Temporal Variation in Humpback Whale (Megaptera novaeangliae) Demographics Along the Western Antarctic Peninsula

Humpback whale populations in Antarctica are recovering after intensecommercial whaling in the 20th century. Along the Western Antarctic Peninsula (WAP)this recovery is occurring in an environment that is experiencing the fastest warming ofany region on the planet. To begin to understand the dynamics of this recovery undersuch dramatic climate change, we are studying the demography of these whales. To date,we have collected 583 biopsy samples from 239 individual males and 268 individualfemales during the austral feeding season from 2010, 2013-2016. The overall sex ratio ofour sample population is 0.89 M:F, supporting early observations that sexes mixrandomly on the feeding grounds. Additionally, we did document a significant seasonalincrease in the proportion of females along the WAP into the fall. We believe that thisshift represents a tendency for pregnant female humpback whales to depart last from thefeeding grounds. Furthermore, we examined progesterone levels of females to assign apregnancy status; providing to our knowledge, the first non-lethal estimation ofpregnancy rates in Antarctic whales. A series of female humpback whales of knownpregnancy status (n=29) from the Northwestern Atlantic, verified from field observations,were used as control samples to develop a logistic regression model, modelling theprobability of pregnancy relative to blubber progesterone concentrations. A pregnancystate was then assigned to females biopsied along the WAP by modelling theirprobability of being pregnant across the control model. Based on our assignment offemales as pregnant not-pregnant, mean progesterone levels for pregnant assignedhumpback whales from the WAP was 250 ng progesterone g blubber (n = 155). Themean value for not-pregnant assigned females was 2.10 ng progesterone g blubber (n =89). Pregnancy rates varied significantly across all years, from 36% in 2010 to 86% in2014.We detected a significant increase in the proportion of pregnant females (58% to72%) from summer to autumn across all years. Some female whales in this populationappear to experience a post-partum ovulation followed by conception (annualpregnancy); on average, more than half (52%) of female whales accompanied by calveswere pregnant. These are some of the first quantitative observations of the demographyof recovering humpback whale populations in the Antarctic and provides a criticalreference point as the Antarctic climate continues to change and populations recover fromwhaling.Keywords: Whaling, Biopsy, Humpback whale, Pregnancy, Progesterone, Climate change, Western Antarctic Peninsula, Sex ratio, Demograph

Validation of a blubber-based endocrine pregnancy test for humpback whales.

Baleen whales have few identifiable external indicators of pregnancy state, making it challenging to study essential aspects of their biology and population dynamics. Pregnancy status in other marine mammals has been determined by measuring progesterone concentrations from a variety of sample matrices, but logistical constraints have limited such studies in free-swimming baleen whales. We use an extensive blubber sample archive and associated calving history data to retrospectively identify samples that correspond to pregnant females and develop a progesterone-based pregnancy test for humpback whales. The lowest pregnant blubber progesterone concentration was 54.97 ng g-1, and the mean for the known-pregnant group was 198.74 ± 180.65 ng g-1. Conversely, females known to be below the minimum age of sexual maturity (juvenile females) had an overall low mean progesterone concentration (0.59 ± 0.25 ng g-1), well below the known-pregnant range. Of the mature females that did not return with a calf (n = 11), three fell within the known-pregnant range (320.79 ± 209.34 ng g-1), while the levels for the remaining eight were two orders of magnitude below the lowest known-pregnant level (1.63 ± 1.15 ng g-1). The proportion of females that did not return with a calf but had values similar to known-pregnant females are consistent with rates of calf mortality, but other potential explanations were considered. Our findings support a validated blubber endocrine assignment of pregnancy corroborated with field life history information, a first for any baleen whale species. The progesterone values we measured were similar to those found in different pregnancy states of other cetaceans and support using blubber biopsy samples for assigning pregnancy in humpback whales. This method can be applied to existing archives or new samples to better study life history and population demography broadly across species and populations

High pregnancy rates in humpback whales (Megaptera novaeangliae) around the Western Antarctic Peninsula, evidence of a rapidly growing population.

Antarctic humpback whales are recovering from near extirpation from commercial whaling. To understand the dynamics of this recovery and establish a baseline to monitor impacts of a rapidly changing environment, we investigated sex ratios and pregnancy rates of females within the Western Antarctic Peninsula (WAP) feeding population. DNA profiling of 577 tissue samples (2010-2016) identified 239 males and 268 females. Blubber progesterone levels indicated 63.5% of the females biopsied were pregnant. This proportion varied significantly across years, from 36% in 2010 to 86% in 2014. A comparison of samples collected in summer versus fall showed significant increases in the proportion of females present (50% to 59%) and pregnant (59% to 72%), consistent with demographic variation in migratory timing. We also found evidence of annual reproduction among females; 54.5% of females accompanied by a calf were pregnant. These high pregnancy rates are consistent with a population recovering from past exploitation, but appear inconsistent with recent estimates of WAP humpback population growth. Thus, our results will help to better understand population growth potential and set a current baseline from which to determine the impact of climate change and variability on fecundity and reproductive rates

Demographic and physiological signals of reproductive events in humpback whales on a southwest pacific breeding ground.

The field of marine mammal conservation has dramatically benefited from the rapid advancement of methods to assess the reproductive physiology of individuals and populations from steroid hormones isolated from minimally invasive skin-blubber biopsy samples. Historically, this vital information was only available from complete anatomical and physiological investigations of samples collected during commercial or indigenous whaling. Humpback whales (Megaptera novaeangliae) are a migratory, cosmopolitan species that reproduce in warm, low-latitude breeding grounds. New Caledonia is seasonally visited by a small breeding sub-stock of humpback whales, forming part of the endangered Oceania subpopulation. To better understand the demographic and seasonal patterns of reproductive physiology in humpback whales, we quantified baseline measurements of reproductive hormones (progesterone-P4, testosterone-T and 17β-estradiol-E2) using an extensive archive of skin-blubber biopsy samples collected from female humpback whales in New Caledonia waters between 2016 and 2019 (n = 194). We observed significant differences in the P4, T and E2 concentrations across different demographic groups of female humpback whales, and we described some of the first evidence of the endocrine patterns of estrous in live free-ranging baleen whales. This study is fundamental in its methodological approach to a wild species that has a global distribution, with seasonally distinct life histories. This information will assist in monitoring, managing and conserving this population as global ecological changes continue to occur unhindered