Forward masking in a bottlenose dolphin Tursiops truncatus : Dependence on azimuthal positions of the masker and test sources

Forward masking was investigated by the auditory evoked potentials (AEP) method in a bottlenose dolphin Tursiops truncatus using stimulation by two successive acoustic pulses (the masker and test) projected from spatially separated sources. The positions of the two sound sources either coincided with or were symmetrical relative to the head axis at azimuths from 0 to ±90°. AEPs were recorded either from the vertex or from the lateral head surface next to the auditory meatus. In the last case, the test source was ipsilateral to the recording side, whereas the masker source was either ipsi- or contralateral. For lateral recording, AEP release from masking (recovery) was slower for the ipsi- than for the contralateral masker source position. For vertex recording, AEP recovery was equal both for the coinciding positions of the masker and test sources and for their symmetrical positions relative to the head axis. The data indicate that at higher levels of the auditory system of the dolphin, binaural convergence makes the forward masking nearly equal for ipsi- and contralateral positions of the masker and test.

Saliva and Blood Cortisol Measurement in Bottlenose Dolphins (Tursiops truncatus): Methodology, Application, and Limitations

A central task of zoos and aquaria is the frequent and accurate assessment of their animals’ welfare. Recently, important steps have been made, such as the introduction of animal welfare evaluation tools and welfare decision trees. To determine animal welfare, it is not only important to collect life history data, such as longevity and reproductive success, but also for experienced observers or caretakers to conduct behavioral observations on a regular basis to assess animals’ emotional state. To physiologically validate welfare observations, glucocorticoid levels are usually assessed, as they are a common indicator of stress. While, for many animals, these levels can be easily determined via fecal or hair samples, for cetaceans, the levels are usually determined via blood samples. As blood samples cannot be taken very frequently and the process may cause stress to the animals (if the samples are not taken following medical training), other techniques, such as the measurement of health biomarkers (especially cortisol, which can be measured in saliva), have become the focus of cetacean stress research. However, there are two problems associated with saliva measurements in cetaceans: saliva might either be diluted with pool water or be contaminated by fodder fish, as frozen fish usually contains high levels of cortisol. In our study, we investigated how saliva cortisol levels are connected to blood cortisol levels and how saliva cortisol can be influenced by fodder fish. We examined saliva and blood samples in eleven bottlenose dolphins (Tursiops truncatus) kept in an outdoor and indoor facility in Germany. Furthermore, we assessed the cortisol levels of different kinds of fodder fish. Our data show that, although saliva cortisol values are elevated under stress and arousal, they seem not to be correlated with blood cortisol values. We also show that, after feeding, saliva cortisol values are increased up to 100-fold. Our results suggest that saliva cortisol measurements in dolphins have to be conducted and considered with care, as they can easily be contaminated. Moreover, it is important to use the right laboratory method in order to specifically detect cortisol; in our study, we conducted reliable tests, using LC-MS/MS.

Cardiac assessments of bottlenose dolphins (Tursiops truncatus) in the Northern Gulf of Mexico following exposure to Deepwater Horizon oil

The Deepwater Horizon (DWH) oil spill profoundly impacted the health of bottlenose dolphins (Tursiops truncatus) in Barataria Bay, LA (BB). To comprehensively assess the cardiac health of dolphins living within the DWH oil spill footprint, techniques for in-water cardiac evaluation were refined with dolphins cared for by the U.S. Navy Marine Mammal Program in 2018 and applied to free-ranging bottlenose dolphins in BB (n = 34) and Sarasota Bay, Florida (SB) (n = 19), a non-oiled reference population. Cardiac auscultation detected systolic murmurs in the majority of dolphins from both sites (88% BB, 89% SB) and echocardiography showed most of the murmurs were innocent flow murmurs attributed to elevated blood flow velocity [1]. Telemetric six-lead electrocardiography detected arrhythmias in BB dolphins (43%) and SB dolphins (31%), all of which were considered low to moderate risk for adverse cardiac events. Echocardiography showed BB dolphins had thinner left ventricular walls, with significant differences in intraventricular septum thickness at the end of diastole (p = 0.002), and left ventricular posterior wall thickness at the end of diastole (p = 0.033). BB dolphins also had smaller left atrial size (p = 0.004), higher prevalence of tricuspid valve prolapse (p = 0.003), higher prevalence of tricuspid valve thickening (p = 0.033), and higher prevalence of aortic valve thickening (p = 0.008). Two dolphins in BB were diagnosed with pulmonary arterial hypertension based on Doppler echocardiography-derived estimates and supporting echocardiographic findings. Histopathology of dolphins who stranded within the DWH oil spill footprint showed a significantly higher prevalence of myocardial fibrosis (p = 0.003), regardless of age, compared to dolphins outside the oil spill footprint. In conclusion, there were substantial cardiac abnormalities identified in BB dolphins which may be related to DWH oil exposure, however, future work is needed to rule out other hypotheses and further elucidate the connection between oil exposure, pulmonary disease, and the observed cardiac abnormalities.

Characterization of Circulating Androgens, Cortisol and Estrogens During Normal, Abnormal and False Pregnancy in Bottlenose Dolphins (Tursiops truncatus) Under Managed Care

The few hormone studies on bottlenose dolphin (Tursiops truncatus) pregnancy with different reproductive outcomes, e.g., normal birth, stillbirth and abortion, have mostly focused on progestagens or relaxin. However, recent analysis of androgens, glucocorticoids and estrogens has shown they are also biomarkers of cetacean pregnancy. Therefore, our objective was to examine circulating concentrations of androgens, glucocorticoids and estrogens during bottlenose dolphin pregnancies with different reproductive outcomes, including normal pregnancy (NORM, n = 27), failure to thrive (FTT, n = 17), perinatal loss (PNL, n = 20), early loss (EL, n = 12) and false pregnancy (FP, n = 16), to determine if they could be potential indicators of reproductive or fetal health. We analyzed longitudinal serum samples (n = 654) from 57 bottlenose dolphins and 92 reproductive events for testosterone, androstenedione, cortisol, estradiol and estrone conjugates. Testosterone concentrations were higher during EL compared to NORM and lower during FP at MID (day 121 – 240 post ovulation/conception) and LATE (day 241 – end of FP) stages (months post conception/ovulation [MPC, MPO] seven through ten, P < 0.05). During FTT, androstenedione concentrations were increased compared to NORM pregnancies in the EARLY and LATE stages (P ≤ 0.05), and concentrations were reduced during FP (P < 0.05). For cortisol, FTT pregnancies had higher concentrations compared to NORM during all stages (P < 0.05), while PNL had higher cortisol during EARLY and LATE stages (P < 0.05). Estradiol concentrations were lower for EL and FP compared to NORM (P < 0.05), while estrone conjugates were only reduced during FP (P < 0.05). Based on our results only cortisol may be a useful predictor of PNL, while both cortisol and androstenedione were useful for distinguishing FTT pregnancies. Similarly, both testosterone and estradiol during EL and FP were different from NORM. Our data indicate a suite of pregnancy specific hormone biomarkers to evaluate maternal and fetal health in bottlenose dolphins should include cortisol, androgens and estrogens. This research also highlights the importance on non-progestagen hormones as sentinels of cetacean pregnancy and fetal health.

Salinity and Water Temperature as Predictors of Bottlenose Dolphin (Tursiops truncatus) Encounter Rates in Upper Galveston Bay, Texas

Bottlenose dolphins (Tursiops truncatus) that inhabit urban estuaries like Galveston Bay, Texas, are exposed to cumulative stressors including pollution, fisheries, shipping, freshwater inflows, and construction operations. With continuing development, it is imperative to understand the key environmental variables that make the Galveston Bay estuary suitable habitat for this protected species. The Galveston Bay Dolphin Research Program conducted monthly photo identification surveys of bottlenose dolphins in a previously understudied 186 km2 area in upper Galveston Bay (UGB). To understand occurrence patterns in this region, we calculated monthly encounter rates of dolphins (dolphins/km) for four consecutive years (2016–2019). Using multiple linear regression models, we investigated the relationship between encounter rates, and water temperature and salinity. Monthly encounter rates ranged from 0.00 to 1.23 dolphins/km with an average of 0.34 dolphins/km (SE = 0.05). Over 80% of the variance was explained by the predictor variables water temperature and salinity (R2 = 0.820). Water temperature had a positive linear effect on encounter rates at over 23.37°C (SE = 1.42). Accordingly, higher encounter rates occurred during months with warm temperatures (May–September) compared to cooler months (November–April), indicating a predictable yearly movement pattern. Moreover, salinity was a highly significant predictor variable, with encounter rates dropping linearly with decreases in salinity. Higher numbers of dolphins are found in UGB during summer, but an exodus of dolphins occurs with low salinity levels, regardless of the time of year and water temperature. These findings should be considered during infrastructure projects (i.e., flood gate system) that may alter dolphin habitat and prey availability.