The significance of blubber hormone sampling in conservation and monitoring of marine mammals

By: Alexa Kownacki, Ph.D. Student, OSU Department of Fisheries and Wildlife, Geospatial Ecology of Marine Megafauna Lab

Marine mammals are challenging to study for many reasons, and specifically because they inhabit the areas of the Earth that are uninhabited by people: the oceans. Monitoring marine mammal populations to gather baselines on their health condition and reproductive status is not as simple as trap and release, which is a method often conducted for terrestrial animals. Marine mammals are constantly moving in vast areas below the surface. Moreover, cetaceans, which do not spend time on land, are arguably the most challenging to sample.

One component of my project, based in California, USA, is a health assessment analyzing hormones of the bottlenose dolphins that frequent both the coastal and the offshore waters. Therefore, I am all too familiar with the hurdles of collecting health data from living marine mammals, especially cetaceans. However, the past few decades have seen major advancements in technology both in the laboratory and with equipment, including one tool that continues to be critical in understanding cetacean health: blubber biopsies.

Biopsy dart hitting a bottlenose dolphin below the dorsal fin. Image Source: NMFS

Blubber biopsies are typically obtained via low-powered crossbow with a bumper affixed to the arrow to de-power it once it hits the skin. The arrow tip has a small, pronged metal attachment to collect an eraser-tipped size amount of tissue with surface blubber and skin. I compare this to a skin punch biopsies in humans; it’s small, minimally-invasive, and requires no follow-up care. With a small team of scientists, we use small, rigid-inflatable vessels to survey the known locations of where the bottlenose dolphins tend to gather. Then, we assess the conditions of the seas and of the animals, first making sure we are collecting from animals without potentially lowered immune systems (no large, visible wounds) or calves (less than one years old). Once we have photographed the individual’s dorsal fin to identify the individual, one person assembles the biopsy dart and crossbow apparatus following sterile procedures when attaching the biopsy tips to avoid infection. Another person prepares to photograph the animal to match the biopsy information to the individual dolphin. One scientist aims the crossbow for the body of the dolphin, directly below the dorsal fin, while the another photographs the biopsy dart hitting the animal and watches where it bounces off. Then, the boat maneuvers to the floating biopsy dart to recover the dart and the sample. Finally, the tip with blubber and skin tissue is collected, again using sterile procedures, and the sample is archived for further processing. A similar process, using an air gun instead of a crossbow can be viewed below:

GEMM Lab members using an air gun loaded with a biopsy dart to procure marine mammal blubber from a blue whale in New Zealand. Video Source: GEMM Laboratory.

Part of the biopsy process is holding ourselves to the highest standards in our minimally-invasive technique, which requires constant practice, even on land.

Alexa practicing proper crossbow technique on land under supervision. Image Source: Alexa Kownacki

Blubber is the lipid-rich, vascularized tissue under the epidermis that is used in thermoregulation and fat storage for marine mammals. Blubber is an ideal matrix for storing lipophilic (fat-loving) steroid hormones because of its high fat content. Steroid hormones, such as cortisol, progesterone, and testosterone, are naturally circulating in the blood stream and are released in high concentrations during specific events. Unlike blood, blubber is less dynamic and therefore tells a much longer history of the animal’s nutritional state, environmental exposure, stress level, and life history status. Blubber is the cribs-notes version of a marine mammal’s biography over its previous few months of life. Blood, on the other hand, is the news story from the last 24 hours. Both matrices serve a specific purpose in telling the story, but blubber is much more feasible to obtain from a cetacean and provides a longer time frame in terms of information on the past.

A simplified depiction of marine mammal blubber starting from the top (most exterior surface) being the skin surface down to the muscle (most interior). Image Source: schoolnet.org.za

I use blubber biopsies for assessing cortisol, testosterone, and progesterone in the bottlenose dolphins. Cortisol is a glucocorticoid that is frequently associated with stress, including in humans. Marine mammals utilize the same hypothalamic-pituitary-adrenal (HPA) axis that is responsible for the fight-or-flight response, as well as other metabolic regulations. During prolonged stressful events, cortisol levels will remain elevated, which has long-term repercussions for an animal’s health, such as lowered immune systems and decreased ability to respond to predators. Testosterone and progesterone are sex hormones, which can be used to indicate sex of the individual and determine reproductive status. This reproductive information allows us to assess the population’s composition and structure of males and females, as well as potential growth or decline in population (West et al. 2014).

Alexa using a crossbow from a small boat off of San Diego, CA. Image Source: Alexa Kownacki

The coastal and offshore bottlenose dolphin ecotypes of interest in my research occupy different locations and are therefore exposed to different health threats. This is a primary reason for conducting health assessments, specifically analyzing blubber hormone levels. The offshore ecotype is found many kilometers offshore and is most often encountered around the southern Channel Islands. In contrast, the coastal ecotype is found within 2 kilometers of shore (Lowther-Thieleking et al. 2015) where they are subjected to more human exposure, both directly and indirectly, because of their close proximity to the mainland of the United States. Coastal dolphins have a higher likelihood of fishery-related mortality, the negative effects of urbanization including coastal runoff and habitat degradation, and recreational activities (Hwang et al. 2014). The blubber hormone data from my project will inform which demographics are most at-risk. From this information, I can provide data supporting why specific resources should be allocated differently and therefore help vulnerable populations. Further proving that the small amount of tissue from a blubber biopsy can help secure a better future for population by adjusting and informing conservation strategies.

Literature Cited:

Hwang, Alice, Richard H Defran, Maddalena Bearzi, Daniela. Maldini, Charles A Saylan, Aime ́e R Lang, Kimberly J Dudzik, Oscar R Guzo n-Zatarain, Dennis L Kelly, and David W Weller. 2014. “Coastal Range and Movements of Common Bottlenose Dolphins (Tursiops Truncatus) off California and Baja California, Mexico.” Bulletin of the Southern California Academy of Sciences 113 (1): 1–13. https://doi.org/10.3390/toxins6010211.

Lowther-Thieleking, Janet L., Frederick I. Archer, Aimee R. Lang, and David W. Weller. 2015. “Genetic Differentiation among Coastal and Offshore Common Bottlenose Dolphins, Tursiops Truncatus, in the Eastern North Pacific Ocean.” Marine Mammal Science 31 (1): 1–20. https://doi.org/10.1111/mms.12135.

West, Kristi L., Jan Ramer, Janine L. Brown, Jay Sweeney, Erin M. Hanahoe, Tom Reidarson, Jeffry Proudfoot, and Don R. Bergfelt. 2014. “Thyroid Hormone Concentrations in Relation to Age, Sex, Pregnancy, and Perinatal Loss in Bottlenose Dolphins (Tursiops Truncatus).” General and Comparative Endocrinology 197: 73–81. https://doi.org/10.1016/j.ygcen.2013.11.021.

Data Wrangling to Assess Data Availability: A Data Detective at Work

By Alexa Kownacki, Ph.D. Student, OSU Department of Fisheries and Wildlife, Geospatial Ecology of Marine Megafauna Lab

Data wrangling, in my own loose definition, is the necessary combination of both data selection and data collection. Wrangling your data requires accessing then assessing your data. Data collection is just what it sounds like: gathering all data points necessary for your project. Data selection is the process of cleaning and trimming data for final analyses; it is a whole new bag of worms that requires decision-making and critical thinking. During this process of data wrangling, I discovered there are two major avenues to obtain data: 1) you collect it, which frequently requires an exorbitant amount of time in the field, in the lab, and/or behind a computer, or 2) other people have already collected it, and through collaboration you put it to a good use (often a different use then its initial intent). The latter approach may result in the collection of so much data that you must decide which data should be included to answer your hypotheses. This process of data wrangling is the hurdle I am facing at this moment. I feel like I am a data detective.

Data wrangling illustrated by members of the R-programming community. (Image source: R-bloggers.com)

My project focuses on assessing the health conditions of the two ecotypes of bottlenose dolphins between the waters off of Ensenada, Baja California, Mexico to San Francisco, California, USA between 1981-2015. During the government shutdown, much of my data was inaccessible, seeing as it was in possession of my collaborators at federal agencies. However, now that the shutdown is over, my data is flowing in, and my questions are piling up. I can now begin to look at where these animals have been sighted over the past decades, which ecotypes have higher contaminant levels in their blubber, which animals have higher stress levels and if these are related to geospatial location, where animals are more susceptible to human disturbance, if sex plays a role in stress or contaminant load levels, which environmental variables influence stress levels and contaminant levels, and more!

Alexa, alongside collaborators, photographing transiting bottlenose dolphins along the coastline near Santa Barbara, CA in 2015 as part of the data collection process. (Image source: Nick Kellar).

Over the last two weeks, I was emailed three separate Excel spreadsheets representing three datasets, that contain partially overlapping data. If Microsoft Access is foreign to you, I would compare this dilemma to a very confusing exam question of “matching the word with the definition”, except with the words being in different languages from the definitions. If you have used Microsoft Access databases, you probably know the system of querying and matching data in different databases. Well, imagine trying to do this with Excel spreadsheets because the databases are not linked. Now you can see why I need to take a data management course and start using platforms other than Excel to manage my data.

A visual interpretation of trying to combine datasets being like matching the English definition to the Spanish translation. (Image source: Enchanted Learning)

In the first dataset, there are 6,136 sightings of Common bottlenose dolphins (Tursiops truncatus) documented in my study area. Some years have no sightings, some years have fewer than 100 sightings, and other years have over 500 sightings. In another dataset, there are 398 bottlenose dolphin biopsy samples collected between the years of 1992-2016 in a genetics database that can provide the sex of the animal. The final dataset contains records of 774 bottlenose dolphin biopsy samples collected between 1993-2018 that could be tested for hormone and/or contaminant levels. Some of these samples have identification numbers that can be matched to the other dataset. Within these cross-reference matches there are conflicting data in terms of amount of tissue remaining for analyses. Sorting these conflicts out will involve more digging from my end and additional communication with collaborators: data wrangling at its best. Circling back to what I mentioned in the beginning of this post, this data was collected by other people over decades and the collection methods were not standardized for my project. I benefit from years of data collection by other scientists and I am grateful for all of their hard work. However, now my hard work begins.

The cutest part of data wrangling: finding adorable images of bottlenose dolphins, photographed during a coastal survey. (Image source: Alexa Kownacki).

There is also a large amount of data that I downloaded from federally-maintained websites. For example, dolphin sighting data from research cruises are available for public access from the OBIS (Ocean Biogeographic Information System) Sea Map website. It boasts 5,927,551 records from 1,096 data sets containing information on 711 species with the help of 410 collaborators. This website is incredible as it allows you to search through different data criteria and then download the data in a variety of formats and contains an interactive map of the data. You can explore this at your leisure, but I want to point out the sheer amount of data. In my case, the OBIS Sea Map website is only one major platform that contains many sources of data that has already been collected, not specifically for me or my project, but will be utilized. As a follow-up to using data collected by other scientists, it is critical to give credit where credit is due. One of the benefits of using this website, is there is information about how to properly credit the collaborators when downloading data. See below for an example:

Example citation for a dataset (Dataset ID: 1201):

Lockhart, G.G., DiGiovanni Jr., R.A., DePerte, A.M. 2014. Virginia and Maryland Sea Turtle Research and Conservation Initiative Aerial Survey Sightings, May 2011 through July 2013. Downloaded from OBIS-SEAMAP (http://seamap.env.duke.edu/dataset/1201) on xxxx-xx-xx.

Citation for OBIS-SEAMAP:

Halpin, P.N., A.J. Read, E. Fujioka, B.D. Best, B. Donnelly, L.J. Hazen, C. Kot, K. Urian, E. LaBrecque, A. Dimatteo, J. Cleary, C. Good, L.B. Crowder, and K.D. Hyrenbach. 2009. OBIS-SEAMAP: The world data center for marine mammal, sea bird, and sea turtle distributions. Oceanography 22(2):104-115

Another federally-maintained data source that boasts more data than I can quantify is the well-known ERDDAP website. After a few Google searches, I finally discovered that the acronym stands for Environmental Research Division’s Data Access Program. Essentially, this the holy grail of environmental data for marine scientists. I have downloaded so much data from this website that Excel cannot open the csv files. Here is yet another reason why young scientists, like myself, need to transition out of using Excel and into data management systems that are developed to handle large-scale datasets. Everything from daily sea surface temperatures collected on every, one-degree of latitude and longitude line from 1981-2015 over my entire study site to Ekman transport levels taken every six hours on every longitudinal degree line over my study area. I will add some environmental variables in species distribution models to see which account for the largest amount of variability in my data. The next step in data selection begins with statistics. It is important to find if there are highly correlated environmental factors prior to modeling data. Learn more about fitting cetacean data to models here.

The ERDAPP website combined all of the average Sea Surface Temperatures collected daily from 1981-2018 over my study site into a graphical display of monthly composites. (Image Source: ERDDAP)

As you can imagine, this amount of data from many sources and collaborators is equal parts daunting and exhilarating. Before I even begin the process of determining the spatial and temporal spread of dolphin sightings data, I have to identify which data points have sex identified from either hormone levels or genetics, which data points have contaminants levels already quantified, which samples still have tissue available for additional testing, and so on. Once I have cleaned up the datasets, I will import the data into the R programming package. Then I can visualize my data in plots, charts, and graphs; this will help me identify outliers and potential challenges with my data, and, hopefully, start to see answers to my focal questions. Only then, can I dive into the deep and exciting waters of species distribution modeling and more advanced statistical analyses. This is data wrangling and I am the data detective.

What people may think a ‘data detective’ looks like, when, in reality, it is a person sitting at a computer. (Image source: Elder Research)

Like the well-known phrase, “With great power comes great responsibility”, I believe that with great data, comes great responsibility, because data is power. It is up to me as the scientist to decide which data is most powerful at answering my questions.

Data is information. Information is knowledge. Knowledge is power. (Image source: thedatachick.com)