Small Blubber Samples (50 mg) Sufficient for Analyses of 10 Stress and Reproductive Steroid Hormones in Gray and Fin Whales via Liquid Chromatography Mass Spectrometry

Information on stress, reproductive fitness, and health is difficult to obtain in wild cetaceans but critical for conservation and management. The goal of this study was to develop a methodology requiring minimal blubber mass for analysis of reproductive and stress steroid hormones and, hence, suitable for cetacean biopsies. Blubber biopsies and samples were collected from free-ranging and stranded gray and fin whales. Steroid hormones were extracted from blubber samples as small as 50 mg using liquid-liquid extraction methodology developed to handle the high fat content of blubber. Samples were analyzed via liquid chromatography with tandem mass spectrometry for 10 hormones: aldosterone, androstenedione, cortisol, cortisone, corticosterone, 17β-estradiol, estrone, 17α-hydroxyprogesterone, progesterone, and testosterone. As part of the optimization, homogenization via bead beating and blade dispersion were compared, and the former found superior. To investigate optimal yet minimal tissue mass required, hormone panels were compared among paired 50, 150, and 400 mg samples, the latter two being commonly reported masses for hormone blubber analysis. Results indicated that 50 mg of blubber was suitable and sometimes superior. Additionally, significant differences in precision values were observed between species, possibly stemming from differences in blubber composition, and relevant to homogenization technique selection and calibration methods that use blubber matrix matches obtained from a species other than the study species. Based on recovery and precision values, our methodology was accurate and precise in the measurement of spiked known quantities for all 10 hormones, confirming the methodology capabilities in 50 mg blubber mass in both species. Altogether, and in our specific sample sets, all endogenous hormones, except corticosterone, were identified above the detection limit in 50 mg gray whale blubber samples while all endogenous hormones, except aldosterone, cortisone, estrone, and progesterone, were detected in 50 mg fin whale blubber samples. We present a robust methodology for the analysis of multiple reproductive and stress steroid hormones in minimal masses of cetacean blubber compatible with small biopsies. Finally, we identified statistically significant differences in corticosteroid concentrations between stranded and free ranging animals.

Source Level of Antarctic Blue and Fin Whale Sounds Recorded on Sonobuoys Deployed in the Deep-Ocean Off Antarctica

The source levels, SL, of Antarctic blue and fin whale calls were estimated using acoustic recordings collected from directional sonobuoys deployed during an Antarctic voyage in 2019. Antarctic blue whale call types included stereotyped song and downswept frequency-modulated calls, often, respectively, referred to as Z-calls (comprising song units-A, B, and C) and D-calls. Fin whale calls included 20 Hz pulses and 40 Hz downswept calls. Source levels were obtained by measuring received levels (RL) and modelling transmission losses (TL) for each detection. Estimates of SL were sensitive to the parameters used in TL models, particularly the seafloor geoacoustic properties and depth of the calling whale. For our best estimate of TL and whale-depth, mean SL in dB re 1 μPa ± 1 standard deviation ranged between 188–191 ± 6–8 dB for blue whale call types and 189–192 ± 6 dB for fin whale call types. These estimates of SL are the first from the Southern Hemisphere for D-calls and 40 Hz downsweeps, and the largest sample size to-date for Antarctic blue whale song. Knowledge of source levels is essential for estimating the detection range and communication space of these calls and will enable more accurate comparisons of detections of these sounds from sonobuoy surveys and across international long-term monitoring networks.

Whaling in Iron Age to post-medieval Scotland

Cetacean remains have been recovered from archaeological sites all over Europe, but are especially abundant in Scotland. These remains originate from all periods and have often been worked into artefacts or tools, including chopping blocks, plaques, combs, pegs, snecks and perforated vertebral epiphyseal discs. It still remains unclear which species were exploited and to what extent active whaling was undertaken in the region. To address these questions Zooarchaeology by Mass Spectrometry (ZooMS) was undertaken on 35 cetacean specimens from five sites in Scotland (Jarlshof, Brough of Birsay, Quoygrew, Deerness and Freswick Links), dating from the Iron Age to the post-medieval period. Furthermore, morphological analysis was performed on the material in order to optimise the ZooMS identifications. A large variety of species were identified, including high numbers of Balaenidae sp and Globicephalinae sp. Comparison with other ZooMS studies in north-western Europe revealed equally high specimen numbers for these species, but also fin whale (Balaenoptera physalus), sperm whale (Phy-seter macrocephalus) and humpback whale (Megaptera novaeangliae). Moreover, one grey whale (Eschrichtius robustus) was identified in the Scottish specimens, adding to an increasing number of specimens indicating that the grey whale was once abundant in European waters. Furthermore, only one specimen of the common minke whale (Balaenoptera acutorostrata) was identified, despite modern stranding data which suggests this is the most common large whale species in Scottish waters. The large variety of species identified suggests that opportunistic scavenging was likely the primary method of acquiring cetaceans, though historical and ethnographic sources suggest that two distinct forms of active whaling may have occasionally been undertaken. The high number of Globicephalinae specimens from Jarlshof raise the possibility that drive-hunting might have already been undertaken at the site during the Iron Age. View supplementary materials here: Table 1 | Table 2 | Table 3 |

The primary visual cortex of Cetartiodactyls: organization, cytoarchitectonics and comparison with perissodactyls and primates

Cetartiodactyls include terrestrial and marine species, all generally endowed with a comparatively lateral position of their eyes and a relatively limited binocular field of vision. To this day, our understanding of the visual system in mammals beyond the few studied animal models remains limited. In the present study, we examined the primary visual cortex of Cetartiodactyls that live on land (sheep, Père David deer, giraffe); in the sea (bottlenose dolphin, Risso’s dolphin, long-finned pilot whale, Cuvier’s beaked whale, sperm whale and fin whale); or in an amphibious environment (hippopotamus). We also sampled and studied the visual cortex of the horse (a closely related perissodactyl) and two primates (chimpanzee and pig-tailed macaque) for comparison. Our histochemical and immunohistochemical results indicate that the visual cortex of Cetartiodactyls is characterized by a peculiar organization, structure, and complexity of the cortical column. We noted a general lesser lamination compared to simians, with diminished density, and an apparent simplification of the intra- and extra-columnar connections. The presence and distribution of calcium-binding proteins indicated a notable absence of parvalbumin in water species and a strong reduction of layer 4, usually enlarged in the striated cortex, seemingly replaced by a more diffuse distribution in neighboring layers. Consequently, thalamo-cortical inputs are apparently directed to the higher layers of the column. Computer analyses and statistical evaluation of the data confirmed the results and indicated a substantial correlation between eye placement and cortical structure, with a markedly segregated pattern in cetaceans compared to other mammals. Furthermore, cetacean species showed several types of cortical lamination which may reflect differences in function, possibly related to depth of foraging and consequent progressive disappearance of light, and increased importance of echolocation.