Apneustic anesthesia ventilation improves pulmonary function in anesthetized bottlenose dolphins (Tursiops truncatus).

Introduction: Use of mechanical ventilation during general anesthesia is a necessary practice in the anesthetization of small cetaceans as spontaneous ventilation fails to provide adequate gas exchange. Currently available methods of ventilation do not account for the intermittent breathing strategy of representative species within this infraorder of fully aquatic mammals and may have a significant effect on cardiac and respiratory physiology. Methods: To understand the impact of mechanical ventilation on cardiopulmonary function in one small species of cetacean, the bottlenose dolphin (Tursiops truncatus), we compared controlled mechanical ventilation (CMV) to a novel ventilation method known as apneustic anesthesia ventilation (AAV). AAV simulates the normal inspiratory breath-hold pattern of dolphins. Ten anesthetic procedures (dental procedure, n = 9; bronchoscopy, n = 2) were performed on nine dolphins (age range: 10-42 years; mean = 32 years; median = 37 years; female = 3, 40%; male = 6, 60%). In a cross-over study design, dolphins were instrumented and randomly assigned to AAV or CMV as the initial mode of ventilation, then switched to the alternate mode. Baseline cardiopulmonary data were collected and again after 30 min on each mode of ventilation. Cardiac index, stroke volume index, systemic vascular resistance, alveolar dead space, alveolar-arterial oxygen tension gradient, arterial oxygen content, oxygen delivery index, and dynamic respiratory system compliance index were calculated at each of the four time points. Results: During AAV, dolphins had higher arterial oxygen tension, higher mean airway pressure, reduced alveolar dead space ventilation and lower alveolar-arterial oxygen difference. Cardiovascular performance was not statistically different between the two modes. Discussion: Our study suggests AAV, which more closely resembles the conscious intermittent respiratory pattern phenotype of dolphins, improves ventilation and pulmonary function in the anesthetized dolphin. Future studies should evaluate the cardiopulmonary effects of neutral buoyancy and cardiopulmonary sparing drug protocols to reduce the need for hemodynamic support of current protocols.

Higher neuron densities in the cerebral cortex and larger cerebellums may limit dive times of delphinids compared to deep-diving toothed whales.

Since the work of Tower in the 1950s, we have come to expect lower neuron density in the cerebral cortex of larger brains. We studied dolphin brains varying from 783 to 6215g. As expected, average neuron density in four areas of cortex decreased from the smallest to the largest brain. Despite having a lower neuron density than smaller dolphins, the killer whale has more gray matter and more cortical neurons than any mammal, including humans. To begin a study of non-dolphin toothed whales, we measured a 596g brain of a pygmy sperm whale and a 2004g brain of a Cuvier's beaked whale. We compared neuron density of Nissl stained cortex of these two brains with those of the dolphins. Non-dolphin brains had lower neuron densities compared to all of the dolphins, even the 6215g brain. The beaked whale and pygmy sperm whale we studied dive deeper and for much longer periods than the dolphins. For example, the beaked whale may dive for more than an hour, and the pygmy sperm whale more than a half hour. In contrast, the dolphins we studied limit dives to five or 10 minutes. Brain metabolism may be one feature limiting dolphin dives. The brain consumes an oversized share of oxygen available to the body. The most oxygen is used by the cortex and cerebellar gray matter. The dolphins have larger brains, larger cerebellums, and greater numbers of cortex neurons than would be expected given their body size. Smaller brains, smaller cerebellums and fewer cortical neurons potentially allow the beaked whale and pygmy sperm whale to dive longer and deeper than the dolphins. Although more gray matter, more neurons, and a larger cerebellum may limit dolphins to shorter, shallower dives, these features must give them some advantage. For example, they may be able to catch more elusive individual high-calorie prey in the upper ocean.

History of the Development of Anesthesia for the Dolphin: A Quest to Study a Brain as Large as Man's.

It is important for academic-minded human anesthesiologists to have an interdisciplinary perspective when engaging in cutting-edge research as well as the practice of human anesthesiology. This was a philosophy promoted by Dr. Robert Dripps, former pioneering Chairman of the Anesthesiology Department at the University of Pennsylvania (Philadelphia, Pennsylvania). Many human and veterinary anesthesiologists as well as biomedical engineers and neuroscientists benefited from Dr. Dripps's constructive outlook personified in the quest to develop dolphin anesthesiology.The motivation to anesthetize dolphins came from the fact that scientists and physicians wanted to study the brain of the dolphin, a brain as large as man's. Also, investigators wanted to develop anesthesia for the dolphin in order to study the electrophysiology of the dolphin's highly sophisticated auditory system, which facilitates the dolphin's amazing echolocation capability.Dolphin anesthesia involves a complex matter of unique neural control, airway anatomy, neuromuscular control of respiration, and sleep behavior.

Timing and context of dolphin clicks during and after mine simulator detection and marking in the open ocean.

Two dolphins carrying cameras swam in the ocean as they searched for and marked mine simulators - buried, proud or moored. As the animals swam ahead of a boat they searched the ocean. Cameras on their harness recorded continuous sound and video. Once a target was detected, the dolphins received a marker to take to the simulator's location. During search and detection, dolphins made almost continuous trains of varying interval clicks. During the marking phase, shorter click trains were interrupted by periods of silence. As the dolphins marked simulators, they often produced victory squeals - pulse bursts that vary in duration, peak frequency and amplitude. Victory squeals were produced on 72% of marks. Sometimes after marking, or at other times during their long swims, the dolphins produced click packets. Packets typically consisted of two to 10 clicks with inter-click intervals of 7-117 ms followed by a silence of 223-983 ms. Click packets appeared unrelated with searching or marking. We suggest that the packets were used to improve signal to noise ratios for locating a boat or other distant object. Victory squeals produced when marking the targets suggest to us that the dolphins know when they have succeeded in this multipart task.

Non-auditory, electrophysiological potentials preceding dolphin biosonar click production.

The auditory brainstem response to a dolphin's own emitted biosonar click can be measured by averaging epochs of the instantaneous electroencephalogram (EEG) that are time-locked to the emitted click. In this study, averaged EEGs were measured using surface electrodes placed on the head in six different configurations while dolphins performed an echolocation task. Simultaneously, biosonar click emissions were measured using contact hydrophones on the melon and a hydrophone in the farfield. The averaged EEGs revealed an electrophysiological potential (the pre-auditory wave, PAW) that preceded the production of each biosonar click. The largest PAW amplitudes occurred with the non-inverting electrode just right of the midline-the apparent side of biosonar click generation-and posterior of the blowhole. Although the source of the PAW is unknown, the temporal and spatial properties rule out an auditory source. The PAW may be a neural or myogenic potential associated with click production; however, it is not known if muscles within the dolphin nasal system can be actuated at the high rates reported for dolphin click production, or if sufficiently coordinated and fast motor endplates of nasal muscles exist to produce a PAW detectable with surface electrodes.

Diffusion tractography reveals pervasive asymmetry of cerebral white matter tracts in the bottlenose dolphin (Tursiops truncatus).

Brain enlargement is associated with concomitant growth of interneuronal distance, increased conduction time, and reduced neuronal interconnectivity. Recognition of these functional constraints led to the hypothesis that large-brained mammals should exhibit greater structural and functional brain lateralization. As a taxon with the largest brains in the animal kingdom, Cetacea provides a unique opportunity to examine asymmetries of brain structure and function. In the present study, diffusion tensor imaging and tractography were used to investigate cerebral white matter asymmetry in the bottlenose dolphin (Tursiops truncatus). Widespread white matter asymmetries were observed with the preponderance of tracts exhibiting leftward structural asymmetries. Leftward lateralization may reflect differential processing and execution of behaviorally variant sensory and motor functions by the cerebral hemispheres. The arcuate fasciculus, an association tract linked to human language evolution, was isolated and exhibited rightward asymmetry suggesting a right hemisphere bias for conspecific communication unlike that of most mammals. This study represents the first examination of cetacean white matter asymmetry and constitutes an important step toward understanding potential drivers of structural asymmetry and its role in underpinning functional and behavioral lateralization in cetaceans.

Comparison of Dolphins' Body and Brain Measurements with Four Other Groups of Cetaceans Reveals Great Diversity.

We compared mature dolphins with 4 other groupings of mature cetaceans. With a large data set, we found great brain diversity among 5 different taxonomic groupings. The dolphins in our data set ranged in body mass from about 40 to 6,750 kg and in brain mass from 0.4 to 9.3 kg. Dolphin body length ranged from 1.3 to 7.6 m. In our combined data set from the 4 other groups of cetaceans, body mass ranged from about 20 to 120,000 kg and brain mass from about 0.2 to 9.2 kg, while body length varied from 1.21 to 26.8 m. Not all cetaceans have large brains relative to their body size. A few dolphins near human body size have human-sized brains. On the other hand, the absolute brain mass of some other cetaceans is only one-sixth as large. We found that brain volume relative to body mass decreases from Delphinidae to a group of Phocoenidae and Monodontidae, to a group of other odontocetes, to Balaenopteroidea, and finally to Balaenidae. We also found the same general trend when we compared brain volume relative to body length, except that the Delphinidae and Phocoenidae-Monodontidae groups do not differ significantly. The Balaenidae have the smallest relative brain mass and the lowest cerebral cortex surface area. Brain parts also vary. Relative to body mass and to body length, dolphins also have the largest cerebellums. Cortex surface area is isometric with brain size when we exclude the Balaenidae. Our data show that the brains of Balaenidae are less convoluted than those of the other cetaceans measured. Large vascular networks inside the cranial vault may help to maintain brain temperature, and these nonbrain tissues increase in volume with body mass and with body length ranging from 8 to 65% of the endocranial volume. Because endocranial vascular networks and other adnexa, such as the tentorium cerebelli, vary so much in different species, brain size measures from endocasts of some extinct cetaceans may be overestimates. Our regression of body length on endocranial adnexa might be used for better estimates of brain volume from endocasts or from endocranial volume of living species or extinct cetaceans.

Evaluation of annual survival and mortality rates and longevity of bottlenose dolphins (Tursiops truncatus) at the United States Navy Marine Mammal Program from 2004 through 2013.

Objective-To evaluate annual survival and mortality rates and the longevity of a managed population of bottlenose dolphins (Tursiops truncatus). Design-Retrospective cohort study. Animals-103 bottlenose dolphins at the US Navy Marine Mammal Program (MMP). Procedures-Population age structures, annual survival and crude mortality rates, and median age at death for dolphins > 30 days old were determined from 2004 through 2013. Results-During 2004 through 2013, the annual survival rates for MMP dolphins ranged from 0.98 to 1.0, and the annual crude mortality rates ranged from 0% to 5%, with a mean of 2.7%. The median age at death was 30.1 years from 2004 through 2008 and increased to 32 years from 2009 through 2013. The maximum age for a dolphin in the study was 52 years. Conclusions and Clinical Relevance-Results indicated that the annual mortality rates were low and survival rates were high for dolphins in the MMP from 2004 through 2013 and that the median age at death for MMP dolphins during that time was over 10 years greater than that reported in free-ranging dolphins. These findings were likely attributable to the continually improving care and husbandry of managed dolphin populations.

The neocortex of cetartiodactyls: I. A comparative Golgi analysis of neuronal morphology in the bottlenose dolphin (Tursiops truncatus), the minke whale (Balaenoptera acutorostrata), and the humpback whale (Megaptera novaeangliae).

The present study documents the morphology of neurons in several regions of the neocortex from the bottlenose dolphin (Tursiops truncatus), the North Atlantic minke whale (Balaenoptera acutorostrata), and the humpback whale (Megaptera novaeangliae). Golgi-stained neurons (n = 210) were analyzed in the frontal and temporal neocortex as well as in the primary visual and primary motor areas. Qualitatively, all three species exhibited a diversity of neuronal morphologies, with spiny neurons including typical pyramidal types, similar to those observed in primates and rodents, as well as other spiny neuron types that had more variable morphology and/or orientation. Five neuron types, with a vertical apical dendrite, approximated the general pyramidal neuron morphology (i.e., typical pyramidal, extraverted, magnopyramidal, multiapical, and bitufted neurons), with a predominance of typical and extraverted pyramidal neurons. In what may represent a cetacean morphological apomorphy, both typical pyramidal and magnopyramidal neurons frequently exhibited a tri-tufted variant. In the humpback whale, there were also large, star-like neurons with no discernable apical dendrite. Aspiny bipolar and multipolar interneurons were morphologically consistent with those reported previously in other mammals. Quantitative analyses showed that neuronal size and dendritic extent increased in association with body size and brain mass (bottlenose dolphin < minke whale < humpback whale). The present data thus suggest that certain spiny neuron morphologies may be apomorphies in the neocortex of cetaceans as compared to other mammals and that neuronal dendritic extent covaries with brain and body size.

Sperm whales and killer whales with the largest brains of all toothed whales show extreme differences in cerebellum.

Among cetaceans, killer whales and sperm whales have the widest distribution in the world's oceans. Both species use echolocation, are long-lived, and have the longest periods of gestation among whales. Sperm whales dive much deeper and much longer than killer whales. It has long been thought that sperm whales have the largest brains of all living things, but our brain mass evidence, from published sources and our own specimens, shows that big males of these two species share this distinction. Despite this, we also find that cerebellum size is very different between killer whales and sperm whales. The sperm whale cerebellum is only about 7% of the total brain mass, while the killer whale cerebellum is almost 14%. These results are significant because they contradict claims that the cerebellum scales proportionally with the rest of the brain in all mammals. They also correct the generalization that all cetaceans have enlarged cerebella. We suggest possible reasons for the existence of such a large cerebellar size difference between these two species. Cerebellar function is not fully understood, and comparing the abilities of animals with differently sized cerebella can help uncover functional roles of the cerebellum in humans and animals. Here we show that the large cerebellar difference likely relates to evolutionary history, diving, sensory capability, and ecology.

Diagnosis and successful treatment of a lung abscess associated with Brucella species infection in a bottlenose dolphin (Tursiops truncatus).

This brief communication describes the clinical presentation, antemortem diagnosis, and successful treatment of a pulmonary abscess associated with a Brucella sp. in a 27-yr-old female bottlenose dolphin (Tursiops truncatus). Ultrasound revealed a 3-cm diameter hypoechoic mass deep to the pleural lining in the left lung field. Multiple ultrasound-guided fine-needle aspirates were performed and tested for bacterial and fungal etiology. All cultures were negative, but the infectious agent was identified by MicroSEQ analysis in two samples and confirmed with real-time polymerase chain reaction (PCR) amplification using known Brucella sp. primers. Amikacin was infused into the abscess and was followed by an oral doxycycline and rifampin protocol. Follow-up diagnostic imaging, including radiographs and computed tomography, revealed a resolved lesion with minimal mineralization within the affected lung fields. Brucellosis should be considered for pulmonary disease in dolphins, and personnel who interact with marine animals should use caution to prevent zoonotic brucellosis.

A mini review of dolphin carbohydrate metabolism and suggestions for future research using exhaled air.

In the 1960s, I explored some aspects of carbohydrate metabolism in healthy bottlenose dolphins (Tursiops truncatus). Their physiological picture resembled what had been described for hyperthyroid diabetics. Dolphins have elevated thyroid hormone turnover, and fasting dolphins maintain a relatively high level of plasma glucose. After dolphins ingest glucose, plasma levels remain high for many hours. Interestingly, plasma glucose must exceed 300 mg/dL (about twice as high as the human threshold) before glucose appears in urine. Due to their diabetes-like states, trainability, and unique natural respiratory anatomy and physiology, dolphins may offer useful clues to metabolites in the breath that may be used to non-invasively monitor diabetes in humans. Dolphins take very rapid and deep breaths that are four or five times as deep as humans and other terrestrial mammals, making them ideal for physiological assessment using non-invasive exhaled air. Avenues for successfully identifying breath-based markers for metabolic disease and physiology in dolphins can be done with both modern technology and the evolutionarily advantageous canine nose. This review summarizes aspects of dolphin metabolism previously learned and offers new directions for diabetes research that may benefit both dolphin and human health.

Dolphins can maintain vigilant behavior through echolocation for 15 days without interruption or cognitive impairment.

In dolphins, natural selection has developed unihemispheric sleep where alternating hemispheres of their brain stay awake. This allows dolphins to maintain consciousness in response to respiratory demands of the ocean. Unihemispheric sleep may also allow dolphins to maintain vigilant states over long periods of time. Because of the relatively poor visibility in the ocean, dolphins use echolocation to interrogate their environment. During echolocation, dolphin produce clicks and listen to returning echoes to determine the location and identity of objects. The extent to which individual dolphins are able to maintain continuous vigilance through this active sense is unknown. Here we show that dolphins may continuously echolocate and accurately report the presence of targets for at least 15 days without interruption. During a total of three sessions, each lasting five days, two dolphins maintained echolocation behaviors while successfully detecting and reporting targets. Overall performance was between 75 to 86% correct for one dolphin and 97 to 99% correct for a second dolphin. Both animals demonstrated diel patterns in echolocation behavior. A 15-day testing session with one dolphin resulted in near perfect performance with no significant decrement over time. Our results demonstrate that dolphins can continuously monitor their environment and maintain long-term vigilant behavior through echolocation.

Neural time and movement time in choice of whistle or pulse burst responses to different auditory stimuli by dolphins.

Echolocating dolphins emit trains of clicks and receive echoes from ocean targets. They often emit each successive ranging click about 20 ms after arrival of the target echo. In echolocation, decisions must be made about the target--fish or fowl, predator or food. In the first test of dolphin auditory decision speed, three bottlenose dolphins (Tursiops truncatus) chose whistle or pulse burst responses to different auditory stimuli randomly presented without warning in rapid succession under computer control. The animals were trained to hold pressure catheters in the nasal cavity so that pressure increases required for sound production could be used to split response time (RT) into neural time and movement time. Mean RT in the youngest and fastest dolphin ranged from 175 to 213 ms when responding to tones and from 213 to 275 ms responding to pulse trains. The fastest neural times and movement times were around 60 ms. The results suggest that echolocating dolphins tune to a rhythm so that succeeding pulses in a train are produced about 20 ms over target round-trip travel time. The dolphin nervous system has evolved for rapid processing of acoustic stimuli to accommodate for the more rapid sound speed in water compared to air.

Evaluation of population health among bottlenose dolphins (Tursiops truncatus) at the United States Navy Marine Mammal Program.

OBJECTIVE: To evaluate health indicators for a population of bottlenose dolphins in the US Navy Marine Mammal Program (MMP) by use of data acquired from 1988 through 2007. DESIGN: Retrospective cohort study. ANIMALS: 167 bottlenose dolphins. PROCEDURES: The following indicators were used to evaluate the health of dolphins during the 20-year period: 5-year age structure, median survival age, annual survival rates, mortality rates, and neonatal and calf survival and mortality rates. Limitations of these population measurements as health indicators for dolphins were assessed. RESULTS: Crude mortality rates of dolphins for 1988 through 1992, 1993 through 1997, 1998 through 2002, and 2003 through 2007 were 3.1%, 4.7%, 3.6%, and 2.4%, respectively; during these same 4 study periods, median survival ages were 14.3, 14.4, 17.7, and 26.1 years, respectively, and mean survival rates were 0.98, 0.97, 0.97, and 0.99, respectively. From 1988 through 1997, 1998 through 2002, and 2003 through 2007, neonatal mortality rates were 4 of 16, 5 of 20, and 2 of 14 neonates, respectively. During these 3 study periods, mean annual survival rates for calves < 3 years old (excluding neonates that died at < 30 days old) were 0.97, 0.92, and 0.99, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Although there were limitations to the measurement of some health indicators, use of multiple methods indicated that the health of dolphins in the MMP population was comparable to, if not better than, that published for other dolphin populations. The MMP population of dolphins may provide useful reference values of health indicators for use in assessment of other managed dolphin populations.

Relationship of blood flow and metabolism to acoustic processing centers of the dolphin brain.

Odontocete brain tissues associated with auditory processing are hypertrophied and modified relative to their terrestrial counterparts. The relationship between the functional demand on these tissues and metabolic substrate requirements is unknown. Using positron emission tomography (PET), relative cerebral blood flow was measured in a bottlenose dolphin. Approximately 60 mCi (13)NH(3) was administered to the dolphin via a catheter inserted into the hepatic vein and threaded proximate to the vena cava. Radiolabel initially appeared as distributed focal points in the cerebellum. Increasing scan time resulted in an increase in the number of focal regions and in the diffusivity of label activity throughout the brain. The time course and spatial distribution of radiolabel was consistent with a cerebral blood supply dominated by the spinal meningeal arteries. Blood flow was predominantly observed in the cerebellum and neocortex, particularly the auditory and visual cortex. Differential brain glucose uptake, previously measured in a separate dolphin, showed good agreement with the differential supply of blood to brain tissues. Rates of blood supply and glucose uptake in the auditory cortex, inferior colliculus, and cerebellum are consistent with a high metabolic demand of tissues which are important to the integration of auditory and other sensory inputs.

A method to enable a bottlenose dolphin (Tursiops truncatus) to echolocate while out of water.

The study of site-specific brain activity associated with dolphin echolocation has been hampered by the difficulties inherent in administering radiolabels and performing medical imaging while a dolphin echolocates in an aquatic environment. To overcome these limitations, a system has been developed to allow a bottlenose dolphin to echolocate while out of the water. The system relies on a "phantom echo generator" (PEG) consisting of a Texas Instruments C6713 digital signal processor with an analog input/output daughtercard. Echolocation clicks produced by the dolphin are detected with a hydrophone embedded in a suction cup on the melon, then digitized within the PEG. Clicks exceeding a user-defined threshold are convolved with a target impulse response, delayed, and scaled before being converted to analog and transmitted through a sound projector embedded in a suction cup attached to the dolphin's lower jaw. Dolphin in-air echolocation behavior, inter-click intervals, and overall performance were analogous to those observed during comparable underwater testing with physical targets, demonstrating that the dolphin was indeed performing an echolocation task while out of water.

Use of phlebotomy treatment in Atlantic bottlenose dolphins with iron overload.

CASE DESCRIPTION: 3 adult (24- to 43-year-old) Atlantic bottlenose dolphins (Tursiops truncatus) with chronic episodic malaise and inappetence associated with high serum aminotransferase (alanine aminotransferase and aspartate aminotransferase) activities, high serum iron concentration, and serum transferrin saturation > 80% were evaluated. CLINICAL FINDINGS: Results of histologic examination of liver biopsy specimens revealed hemosiderosis in all 3 dolphins. Except for chronic lymphocytosis in 1 dolphin, results of extensive diagnostic testing revealed no other abnormalities. For each dolphin, a diagnosis of iron overload of unknown origin was made. TREATMENT AND OUTCOME: Phlebotomy treatment was implemented to reduce body stores of iron. Each phlebotomy procedure removed 7% to 17% (1 to 3 L) of estimated blood volume. Treatment consisted of an induction phase of weekly phlebotomy procedures for 22 to 30 weeks, which was complete when serum iron concentration and aminotransferase activities were within reference ranges and serum transferrin saturation was < or = 20% or Hct was < or = 30%. Total amount of iron removed from each dolphin was 53 to 111 mg/kg (24.1 to 50.5 mg/lb) of body weight. One dolphin required maintenance procedures at 8- to 12-week intervals when high serum iron concentration was detected. CLINICAL RELEVANCE: Although the cause of the iron overload and high serum aminotransferase activities remained unknown, phlebotomy treatment successfully resolved the clinicopathologic abnormalities, supporting a role of iron overload in the hepatopathy of the 3 dolphins.

Cetacean sleep: an unusual form of mammalian sleep.

Our knowledge of the form of lateralized sleep behavior, known as unihemispheric slow wave sleep (USWS), seen in all members of the order Cetacea examined to date, is described. We trace the discovery of this phenotypically unusual form of mammalian sleep and highlight specific aspects that are different from sleep in terrestrial mammals. We find that for cetaceans sleep is characterized by USWS, a negligible amount or complete absence of rapid eye movement (REM) sleep, and a varying degree of movement during sleep associated with body size, and an asymmetrical eye state. We then compare the anatomy of the mammalian somnogenic system with what is known in cetaceans, highlighting areas where additional knowledge is needed to understand cetacean sleep. Three suggested functions of USWS (facilitation of movement, more efficient sensory processing and control of breathing) are discussed. Lastly, the possible selection pressures leading to this form of sleep are examined, leading us to the suggestion that the selection pressure necessitating the evolution of cetacean sleep was most likely the need to offset heat loss to the water from birth and throughout life. Aspects such as sentinel functions and breathing are likely to be proximate evolutionary phenomenon of this form of sleep.