Forward shift of feeding buzz components of dolphins and belugas during associative learning reveals a likely connection to reward expectation, pleasure and brain dopamine activation.

For many years, we heard sounds associated with reward from dolphins and belugas. We named these pulsed sounds victory squeals (VS), as they remind us of a child's squeal of delight. Here we put these sounds in context with natural and learned behavior. Like bats, echolocating cetaceans produce feeding buzzes as they approach and catch prey. Unlike bats, cetaceans continue their feeding buzzes after prey capture and the after portion is what we call the VS. Prior to training (or conditioning), the VS comes after the fish reward; with repeated trials it moves to before the reward. During training, we use a whistle or other sound to signal a correct response by the animal. This sound signal, named a secondary reinforcer (SR), leads to the primary reinforcer, fish. Trainers usually name their whistle or other SR a bridge, as it bridges the time gap between the correct response and reward delivery. During learning, the SR becomes associated with reward and the VS comes after the SR rather than after the fish. By following the SR, the VS confirms that the animal expects a reward. Results of early brain stimulation work suggest to us that SR stimulates brain dopamine release, which leads to the VS. Although there are no direct studies of dopamine release in cetaceans, we found that the timing of our VS is consistent with a response after dopamine release. We compared trained vocal responses to auditory stimuli with VS responses to SR sounds. Auditory stimuli that did not signal reward resulted in faster responses by a mean of 151 ms for dolphins and 250 ms for belugas. In laboratory animals, there is a 100 to 200 ms delay for dopamine release. VS delay in our animals is similar and consistent with vocalization after dopamine release. Our novel observation suggests that the dopamine reward system is active in cetacean brains.

Vocal reporting of echolocation targets: dolphins often report before click trains end.

Bottlenose dolphins (Tursiops truncatus) wore opaque suction cups over their eyes while stationing behind an acoustically opaque door. This put the dolphins in a known position and orientation. When the door opened, the dolphin clicked to detect targets. Trainers specified that Dolphin S emit a whistle if the target was a 7.5 cm water filled sphere, or a pulse burst if the target was a rock. S remained quiet if there was no target. Dolphin B whistled for the sphere. She remained quiet for rock and for no target. Thus, S had to choose between three different responses, whistle, pulse burst, or remain quiet. B had to choose between two different responses, whistle or remain quiet. S gave correct vocal responses averaging 114 ms after her last echolocation click (range 182 ms before and 219 ms after the last click). Average response for B was 21 ms before her last echolocation click (range 250 ms before and 95 ms after the last click in the train). More often than not, B began her whistle response before her echolocation train ended. The findings suggest separate neural pathways for generation of response vocalizations as opposed to echolocation clicks.

Cetacean brain evolution: Dwarf sperm whale (Kogia sima) and common dolphin (Delphinus delphis) - An investigation with high-resolution 3D MRI.

This study compares a whole brain of the dwarf sperm whale (Kogia sima) with that of a common dolphin (Delphinus delphis) using high-resolution magnetic resonance imaging (MRI). The Kogia brain was scanned with a Siemens Trio Magnetic Resonance scanner in the three main planes. As in the common dolphin and other marine odontocetes, the brain of the dwarf sperm whale is large, with the telencephalic hemispheres remarkably dominating the brain stem. The neocortex is voluminous and the cortical grey matter thin but expansive and densely convoluted. The corpus callosum is thin and the anterior commissure hard to detect whereas the posterior commissure is well-developed. There is consistency as to the lack of telencephalic structures (olfactory bulb and peduncle, olfactory ventricular recess) and neither an occipital lobe of the telencephalic hemisphere nor the posterior horn of the lateral ventricle are present. A pineal organ could not be detected in Kogia. Both species show a tiny hippocampus and thin fornix and the mammillary body is very small whereas other structures of the limbic system are well-developed. The brain stem is thick and underlies a large cerebellum, both of which, however, are smaller in Kogia. The vestibular system is markedly reduced with the exception of the lateral (Deiters') nucleus. The visual system, although well-developed in both species, is exceeded by the impressive absolute and relative size of the auditory system. The brainstem and cerebellum comprise a series of structures (elliptic nucleus, medial accessory inferior olive, paraflocculus and posterior interpositus nucleus) showing characteristic odontocete dimensions and size correlations. All these structures seem to serve the auditory system with respect to echolocation, communication, and navigation.

Dolphin lung collapse and intramuscular circulation during free diving: evidence from nitrogen washout.

Intramuscular nitrogen tensions in Tursiops truncatus after a schedule of repetitive ocean dives suggest a lung collapse depth of about 70 meters and suggest that intramuscular circulation is maintained during unrestrained diving in the open ocean. Therefore, the bottle-nosed dolphin is not protected by lung collapse from the decompression hazards of dives to depths shallower than 70 meters.

Blood oxygen and ecology of porpoises of three genera.

Blood volumes, hemoglobin concentrations, packed-cell volumes, and heart weights were determined in three genera of propoises which differ from one another in behavior and ecology. The estimate for the total blood-oxygen content of the highly active, deep-diving, pelagic species Phocoenoides dalli was almost three times greater than that for the coastaldwelling species Tursiops truncatus, and about 70 percent greater than for the less active pelagic species, Lagenorhynchus obliquidens. Heart weights of Phocoenoides dalli were about 140 percent greater than in Tursiops truncatus and 55 percent greater than in Logenorhynchus obliquidens.

Respiration and deep diving in the bottlenose porpoise.

A bottlenose porpoise was trained to dive untethered in the open ocean and to exhale into an underwater collecting funnel before surfacing from prescribed depths down to 300 meters. The animal was also taught to hold its breath for periods up to 4 minutes at the surface and then blow in the funnel. Alveolar collapse is probably complete at around 100 meters, and little pulmonary respiratory exchange occurs below that depth. Thoracic collapse was observ visually at 10 to 50 meters and by underwater television to 300 meters.

Anesthetization of porpoises for major surgery.

Comparison of three porpoises (Tursiops truncatus and Lagenorhynchus obliquidens) given nitrous oxide with 18 given halothane, with complete documentation of reflexes and comprehensive physiological monitoring, showed halothane to be a suitable anesthetic for major surgery while nitrous oxide was found to be inadequate. In addition, sodium thiopental administered intravenously was successfully used to facilitate intubation procedures. This development eliminated the need to intubate awake porpoises.

Sleep and cardiac rhythm in the gray seal.

Telemetric studies of electroencephalograms, electrocardiograms, and electroculograms and concurrent observations of behavior revealed that seals can sleep underwater, on the surface, or while hauled out. Rapid eye movement preceded slow wave sleep and was accompanied by increased respiratory rate and rhythmic tachycardia. While slow wave sleep occurred under all sleep conditions, rapid eye movement occurred only when a seal was hanging at the water surface or hauled out, never underwater.

Morphology and evolutionary biology of the dolphin (Delphinus sp.) brain--MR imaging and conventional histology.

Whole brains of the common dolphin (Delphinus delphis) were studied using magnetic resonance imaging (MRI) in parallel with conventional histology. One formalin-fixed brain was documented with a Siemens Trio Magnetic Resonance scanner and compared to three other brains which were embedded in celloidin, sectioned in the three main planes and stained for cells and fibers. The brain of the common dolphin is large, with the telencephalic hemispheres dominating the brain stem. The neocortex is voluminous and the cortical grey matter thin but extremely extended and densely convoluted. There is no olfactory ventricular recess due to the lack of an anterior olfactory system (olfactory bulb and peduncle). No occipital lobe of the telencephalic hemisphere and no posterior horn of the lateral ventricle are present. A pineal organ could not be detected. The brain stem is thick and underlies a very large cerebellum. The hippocampus and mammillary body are small and the fornix is thin; in contrast, the amygdaloid complex is large and the cortex of the limbic lobe is extended. The visual system is well developed but exceeded by the robust auditory system; for example, the inferior colliculus is several times larger than the superior colliculus. Other impressive structures in the brainstem are the peculiar elliptic nucleus, inferior olive, and in the cerebellum the huge paraflocculus and the very large posterior interpositus nucleus. There is good correspondence between MR scans and histological sections. Most of the brain characteristics can be interpreted as morphological correlates to the successful expansion of this species in the marine environment, which was characterized by the development of a powerful sonar system for localization, communication, and acousticomotor navigation.

Neuron numbers in sensory cortices of five delphinids compared to a physeterid, the pygmy sperm whale.

With its large mass and enormous gyrification, the neocortex of whales and dolphins has always been a challenge to neurobiologists. Here we analyse the relationship between neuron number per cortical unit in three different sensory areas and brain mass in six different toothed whale species, five delphinids and one physeterid. Cortex samples, including primary cortical areas of the auditory, visual, and somatosensory systems were taken from both hemispheres of brains fixed in 10% buffered formalin. The samples were embedded in paraffin, sectioned at 25 microm thickness and stained with cresyl violet. Because cortical thickness varies among toothed whale species, cell counts were done in cortical units measuring 150mum in width, 25 microm in thickness, and extending from the pial surface to the white matter. By arranging the delphinid brains according to their total mass, 834-6052 g, we found decreasing neuron numbers in the investigated areas with increasing brain mass. The pigmy sperm whale (Kogia breviceps), a physeterid with an adult brain weight of 1000 g had a distinctly lower neuron number per cortical unit. As had been expected, an increase in adult brain weight in delphinid cetaceans (family Delphinidae) is not correlated with an increase in neuron number per cortical unit.

Dolphin peripheral visual pathway in chronic unilateral ocular atrophy: complete decussation apparent.

Components of the peripheral visual pathway were examined in two bottlenose dolphins, Tursiops truncatus, each with unilateral ocular degeneration and scarring of 3 or more years' duration. In both animals, the optic nerve associated with the blind eye (right eye in Tg419 and left eye in Tt038) had a translucent, gel-like appearance upon gross examination. This translucency was also evident in the optic tract contralateral to the affected eye. In Tg419, myelinated axons of varying diameters were apparent in the left optic nerve, whereas the right optic nerve, serving the blind eye, appeared to be devoid of axons. In Tt038, myelinated axons were associated with the right optic nerve (serving the functional eye) and left optic tract but were essentially absent in the left optic nerve and right optic tract. Examined by light microscopy in serial horizontal sections, the optic chiasm of Tt038 was arranged along its central plane in segregated, alternating pathways for the decussation of right and left optic nerve fibers. Ventral to this plane, the chiasm was comprised of fibers from the left optic nerve, whereas dorsal to the central plane, fibers derived from the right optic nerve. Because of this architectural arrangement, the right and left optic nerves grossly appeared to overlap as they crossed the optic chiasm with the right optic nerve coursing dorsally to the left optic nerve. At the light and electron microscopic levels, the optic nerves and tracts lacking axons were well vascularized and dominated by glial cell bodies and glial processes, an expression of the marked glial scarring associated with postinjury axonal degeneration. The apparent absence of axons in one of the optic tract pairs (right in Tt038 and left in Tg419) supports the concept of complete decussation of right and left optic nerve fibers at the optic chiasm in the bottlenose dolphin.

Opportunities for using Navy marine mammals to explore associations between organochlorine contaminants and unfavorable effects on reproduction.

The Department of Defense (DoD) has a unique marine mammal program maintained by the US Navy that includes the largest force of bottlenose dolphins, Tursiops truncatus, worldwide. In recent years, this population of cetaceans that lives in netted open water enclosures in San Diego Bay has been monitored for levels of organochlorine (OC) contaminants in blubber, blood and milk. Data generated from these studies have afforded insight into the fate and possible effects of OC contaminants in marine mammals. We now report preliminary findings on the effects of maternal OC exposure on pregnancy outcome. Blubber OC levels were compared between females whose calves survived beyond 6 months and females whose calves were stillborn or died within 12 days of birth. The mean concentration of SigmaDDT was more than 3 times as high among dolphins whose calves died as that among dolphins whose calves survived beyond 6 months (P = 0.002). Mean SigmaPCB was more than 2.5 times higher in females whose calves did not survive (P= 0.076). This population is a logical sentinel for the assessment of environmentally mediated disease. Biological tissues and fluids can be sampled on a regular basis from the dolphins for accumulation of tissue residues, facilitated by conditioned husbandry behaviors. These trained behaviors help preclude possible alterations in health measures resulting from capture stress. Animals' diets can be monitored for contaminant levels. With these data, the expertise and facilities available at the Navy laboratory and in collaboration with other experts in the field, controlled studies can be designed to monitor and assess dietary exposure, measurable immune and neurologic responses and assess reproductive and transgenerational effects of contaminants. Biomarkers can be developed to relate the health of individual animals relative to contaminant exposures. Such investigations of natural exposure and response scenarios are a logical adjunct to traditional laboratory toxicity studies.

Reported causes of death of captive killer whales (Orcinus orca).

Inquiries were made to all oceanaria that maintain killer whales in North America. Causes of death determined at necropsy included mediastinal abscesses, pyometra, pneumonia, influenza, salmonellosis, nephritis, Chediak-Higashi syndrome, fungus infection, ruptured aorta, cerebral hemorrhage and a perforated post-pyloric ulcer. Captive females appear to have a higher rate of mortality than males. Growth rates for whales that died were greater than for those that survived.

A trematode from the round window of an Atlantic Bottlenosed dolphin's ear.

A trematode from the family Nasitrematidae Yamaguti 1951 was found adhered to the round window in the inner ear of a Bottlenosed Dolphin (Tursiops truncatus). The possibility that parasites could be responsible for changes in acoustic behavior and hearing loss is discussed.

Mandibular anesthesia and tooth extraction in the bottlenosed dolphin.

Anatomical dissections were done to show the innervation of the teeth and mandible of the bottlenosed dolphin (Turslops truncatus). Using structural landmarks, a method has been devised for anesthetizing the lower jaw. With this procedure teeth can be extracted and age determined by counting dentine layers in sections of etched teeth. Animals of the most desirable ages can thus be selected and the ages of animals already in captivity can be determined.

Weight-length relationships of wild-caught and captive Atlantic bottlenose dolphins.

Length and weight measurements were obtained on 144 Atlantic bottlenose dolphins (Tursiops truncatus). Data were compared for wild-caught dolphins (n = 124), dolphins in training (n = 34), and dolphins that had died in an emaciated condition (n = 15). The purpose of the study was to establish guidelines for use in estimating when a dolphin is over- or underweight. Regression lines were plotted to give the minimal acceptable weight for any length from 185 to 265 cm.