Lorrie D. Rea

We evaluated the Precision Xtra™ ketometer as part of a larger study categorizing fasting status of free-ranging Steller sea lion (Eumetopias jubatus; SSL) pups which necessitated the identification of plasma β-hydroxybutyrate concentrations ([β-HBA]) around a threshold of n = 14; triplicate technical replicates). Plasma (stored at −80°C, NaHep, Thaw1) measured via our laboratory’s Reference Assay (Sigma Aldrich, St. Louis, MO, Kit #MAK041) served as the standard [β-HBA] for ketometer comparisons. Our observed β-HBA range (0.0–1.6 mmol/l), consistent with published [β-HBA] of free-ranging Otariid pups, represented the lower 20% of the ketometer’s range (0.0–8.0 mmol/l). The maximal coefficient of variation (%CV) of ketometer technical replicates was 9.1% (NaHep, whole blood). The majority of ketometer technical replicate sets (84%, including all matrices, anticoagulants and thawings) were identical (CV = 0%). We found linear relationships and agreement of ketometer [β-HBA] between whole blood preserved with different anticoagulants and between whole blood and plasma (Thaw1) measurements. The ketometer produced results with linearity to the Reference Assay for both whole blood and plasma (Thaw1). We identified a non-linear relationship between plasma at Thaw1 and Thaw2 (tested four months apart, NaHep), as only samples with higher SSL [β-HBA] decreased in concentration, and all others remained the same. With respect to categorizing SSL pup fasting in our larger study, the ketometer’s %Accuracy, %Sensitivity and %Specificity for samples with Reference Assay β-HBA 0.4 mmol/l were 100%. We adopted a modified procedure: plasma samples with mean ketometer concentrations ±0.1 mmol/l of 0.3 mmol/l β-HBA were re-evaluated using the Reference Assay, improving measurement precision from tenths (ketometer) to thousandths (assay) mmol/l. The Precision Xtra™ ketometer was valuable to our application over the range of [β-HBA] observed in SSL pup plasma and whole blood samples.

Geographic differences in population growth trends are well-documented in Steller sea lions (Eumetopias jubatus), a species of North Pacific pinniped listed under the U.S. Endangered Species Act in 1990 following a marked decline in population abundance that began during the 1970s. As population growth is intrinsically linked to pup production and survival, examining factors related to pup physiological condition provides useful information to management authorities regarding potential drivers of regional differences. During dam foraging trips, pups predictably transition among three fasting phases, distinguished by the changes in the predominant metabolic byproduct. We used standardized ranges of two plasma metabolites (blood urea nitrogen and β–hydroxybutyrate) to assign pups to fasting categories (n = 1528, 1990–2016, 12 subpopulations): Recently Fed–Phase I (digestion/assimilation–expected hepatic/muscle glycogen usage), Phase II (expected lipid utilization), transitioning between Phases II–III (expected lipid utilization with increased protein reliance), or Phase III (expected protein catabolism). As anticipated, the majority of pups were classified as Recently Fed–Phase I (overall mean proportion = 0.72) and few pups as Phase III (overall mean proportion = 0.04). By further comparing pups in Short (Recently Fed–Phase II) and Long (all other pups) duration fasts, we identified three subpopulations with significantly (P < 0.03) greater proportions of pups dependent upon endogenous sources of energy for extended periods, during a life stage of somatic growth and development: the 1) central (0.27 ± 0.09) and 2) western (0.36 ± 0.13) Aleutian Island (declining population trend) and 3) southern Southeast Alaska (0.32 ± 0.06; increasing population trend) subpopulations had greater Long fast proportions than the eastern Aleutian Islands (0.10 ± 0.05; stabilized population). Due to contrasting population growth trends among these highlighted subpopulations over the past 50+ years, both density-independent and density-dependent factors likely influence the dam foraging trip duration, contributing to longer fasting durations for pups at some rookeries.

Steller sea lion (Eumetopias jubatus) young‐of‐the‐year (YOY) are nutritionally dependent upon their dam through the majority of their first year. Several indices of body condition were measured in YOY 1.5–9 mo of age captured in Southeast Alaska (n = 122), the Gulf of Alaska (n = 182), and the Aleutian Islands (n = 38) to test the hypothesis that nutritional stress impacted the ability of adult female Steller sea lions to adequately nourish their late gestation YOY in the central Aleutian Islands in the early 2000s. Body mass (kg) and percent total body lipid content (%TBL) increased with age in all three regions of Alaska that were sampled (P < 0.05). Young‐of‐the‐year 7–9 mo of age were leaner in Southeast Alaska (27.6% ± 1.0%) and Gulf of Alaska (29.5% ± 0.8%) than in the Aleutian Islands (35.7% ± 1.2%, P < 0.001). Condition indices calculated from morphometric measures did not strongly predict the %TBL measured by isotope dilution. The trend for Aleutian Island YOY to have larger body mass and larger body fat reserves are counter to what would be expected if dams were unable to adequately provision their late lactation YOY due to inadequate food availability in the central Aleutian Islands.

Steller sea lions (SSL; Eumetopias jubatus) grow their vibrissae continually, providing a multiyear record suitable for ecological and physiological studies based on stable isotopes. An accurate age‐specific vibrissae growth rate is essential for registering a chronology along the length of the record, and for interpreting the timing of ecologically important events. We utilized four methods to estimate the growth rate of vibrissae in fetal, rookery pup, young‐of‐the‐year (YOY), yearling, subadult, and adult SSL. The majority of vibrissae were collected from SSL live‐captured in Alaska and Russia between 2000 and 2013 (n = 1,115), however, vibrissae were also collected from six adult SSL found dead on haul‐outs and rookeries during field excursions to increase the sample size of this underrepresented age group. Growth rates of vibrissae were generally slower in adult (0.44 ± 0.15 cm/mo) and subadult (0.61 ± 0.10 cm/mo) SSL than in YOY (0.87 ± 0.28 cm/mo) and fetal (0.73 ± 0.05 cm/mo) animals, but there was high individual variability in these growth rates within each age group. Some variability in vibrissae growth rates was attributed to the somatic growth rate of YOY sea lions between capture events (P = 0.014, r² = 0.206, n = 29).

Knowledge of diet–tissue stable isotope discrimination is required to properly interpret stable isotope values and to identify possible diet shifts, such as might be expected from nursing through weaning. This study compared δ¹³C and δ¹⁵Ν οf paired serum and vibrissal roots with those of ingested milk (n = 52) from free-ranging Steller sea lion ( Eumetopias jubatus (Schreber, 1776)) pups (1–11 months) and juveniles (14–27 months) to estimate diet–tissue discrimination. Mean ¹⁵N enrichment from ingested milk to serum was 2.1‰ ± 0.6‰ and δ¹⁵Ν at the root of the vibrissae (representing current growth) were not significantly different from serum values. Milk was enriched for mean ¹³C by 5.0‰ ± 1.0‰ and 7.3‰ ± 1.2‰ relative to serum and vibrissal roots, respectively, which was due to the presence of ¹³C-depleted lipids in milk. This was confirmed by lipid extraction from a subset of milk and serum samples, resulting in a 5.8‰ ± 1.0‰ change only in milk. This study established that vibrissal roots and serum are reflective of a milk diet with approximately 2.0‰ ¹⁵N enrichment, and vibrissal roots reflect serum and lipid-extracted milk values with approximately 2.0‰ ¹³C enrichment. These discrimination factors are important to establish for stable isotope studies assessing diet shifts.