Iron Indices in Bottlenose Dolphins (Tursiops truncatus)

Bottlenose dolphins can have iron overload (that is, hemochromatosis), and managed populations of dolphins may be more susceptible to this disease than are wild dolphins. Serum iron, total iron-binding capacity (TIBC), transferrin saturation, and ferritin were measured in 181 samples from 141 dolphins in 2 managed collections and 2 free-ranging populations. Although no iron indices increased with age among free-ranging dolphins, ferritin increased with age in managed collections. Dolphins from managed collections had higher iron, ferritin, and transferrin saturation values than did free-ranging dolphins. Dolphins with high serum iron (exceeding 300 μg/dL) were more likely to have elevated ferritin but not ceruloplasmin or haptoglobin, demonstrating that high serum levels of iron are due to a true increase in total body iron. A time-series study of 4 dolphins with hemochromatosis that were treated with phlebotomy demonstrated significant decreases in serum ferritin, iron, and TIBC between pre- and posttreatment samples; transferrin saturation initially fell but returned to prephlebotomy levels by 6 mo after treatment. Compared with those in managed collections, wild dolphins were 15 times more likely to have low serum iron (100 μg/dL or less), and this measure was associated with lower haptoglobin. In conclusion, bottlenose dolphins in managed collections are more likely to have greater iron stores than are free-ranging dolphins. Determining why this situation occurs among some dolphin populations and not others may improve the treatment of hemochromatosis in dolphins and provide clues to causes of nonhereditary hemochromatosis in humans.Abbreviations: TIBC, total iron-binding capacityBottlenose dolphins (Tursiops truncatus) are susceptible to iron overload (that is, hemochromatosis).^19,32,33,35 Dolphins with hemochromatosis have high serum iron levels that progress over years, have high transferrin saturation (85% and greater), and are responsive to phlebotomy therapy.¹⁹ Left untreated, dolphins with hemochromatosis are more likely to have increased liver aminotransferases, chronic inflammation, and hyperlipidemia than are healthy controls.³⁵ A 25-y retrospective study of one population demonstrated that 67% of dolphins had excessive hepatic hemosiderin deposition at time of death, 92% of which had hemosiderin deposition in Kupffer cells; hemolytic anemia, anemia of chronic disease, and viral infections were not associated with hemosiderin deposition, and the primary hypothesis is that dolphins in managed collections may be susceptible to iron storage disease.³²The etiology of dolphin iron overload is unknown. The most common cause in humans is hereditary hemochromatosis, which is most often attributed to a point mutation of the HFE gene.¹² Other causes of iron overload include primary liver disease, excessive iron intake, and insulin resistance.^3,36 Comparisons of the HFE gene among dolphins with and without hemochromatosis showed no significant differences (unpublished data), and iron deposition in Kupffer cells (compared with hepatocytes) is not supportive of a hereditary cause.^3,32 Liver enzymes decrease in response to the removal of body iron through phlebotomy; this effect indicates that iron overload leads to liver disease rather than that primary liver disease causes iron overload.¹⁹ At least one dolphin population from which cases of hemochromatosis have been reported have not received iron supplementation for more than 10 y, although the iron content in their frozen–thawed fish diet has not been assessed fully.¹⁹ Dolphins with hemochromatosis have significantly higher 2-h postprandial insulin levels than do controls, suggesting that insulin resistance plays a role in this disease, as occurs in humans.³³In addition to dolphins and humans, iron overload occurs in diverse captive birds, callitrichids, black rhinos, Egyptian fruit bats, lemurs, northern fur seals, California sea lions, tapirs, and Saler cattle.^{5,11,15,18,20,21,23,29,31} Similar to that in other exotic species, iron overload in dolphins is thought to be a disease that is associated with managed care, but comparisons of iron levels among managed collections and free-ranging dolphin populations have been lacking.¹⁹ Further, measurements of serum ferritin, ceruloplasmin, and haptoglobin have not typically been included in iron overload evaluations in dolphins. Although serum ferritin levels can be an indicator of total iron body stores, ferritin also increases in response to inflammation or hemolytic anemia.⁴ In humans, ceruloplasmin is a plasma protein that increases with acute and chronic inflammation and plays a role in iron metabolism. Haptoglobin is a protein that, in humans, decreases with intravascular hemolytic anemia, among other conditions.² As such, concurrent measurements of iron, ferritin, ceruloplasmin, and haptoglobin provide insight into the true cause of changing iron and ferritin levels.Low serum iron is used as a general indicator of inflammation in marine mammals, and return of serum iron to normal levels during illness is seen as a positive indicator for recovery.^13,24 The dynamics of iron, erythrocyte sedimentation rate, and fibrinogen in inflammation have remained important as indicators of dolphin health for decades, yet little has been published about dolphin iron stores and associated acute-phase proteins in the past 30 y.²⁴ One barrier to the use of acute-phase proteins, including C-reactive protein, has been the need for species-specific assays; development of tests that better detect early phases of inflammation and infection could lead to earlier treatment and better outcomes. Other barriers include the need for consecutive samples from dolphins with known health status to validate indicators for inflammation, including ceruloplasmin.To better understand the prevalence of high serum iron and potential risks for hemochromatosis among various dolphin populations, indices of serum iron, ferritin, total iron-binding capacity (TIBC), percentage transferrin saturation, ceruloplasmin, and haptoglobin concentrations were measured in 2 populations of free-ranging dolphins and 2 managed collections, including the cohort of Navy dolphins. These variables were compared by age, sex, and population. Serum ferritin was measured by using a newly developed dolphin-specific assay.     

Toxoplasmosis in Atlantic bottle-nosed dolphins (Tursiops truncatus)

A female Atlantic bottle-nosed dolphin (Tursiops truncatus) and her calf were found beached on Picnic Island in Tampa Bay, Florida, USA. Despite therapy the animals died. Necropsy revealed severe pneumonia and lymphadenopathy in the mother and the calf, gastric ulcers and infection with the stomach digenean Braunina cordiformis in the mother, and a large, pale liver in the calf. Toxoplasma gondii was identified by light and electron microscopy and by immunohistochemistry in tissues of both animals. Toxoplasma gondii was associated with interstitial pneumonia, necrotizing adrenalitis, and cardiac myonecrosis in the mother and with lymphoid necrosis in both dolphins. The source of infection and the relationship to the recent dolphin beachings along the eastern seacoast of North America are unknown. This is the first report of toxoplasmosis in cetaceans.     

Signature-whistle production in undisturbed free-ranging bottlenose dolphins (Tursiops truncatus)

Data from behavioural observations and acoustic recordings of free-ranging bottlenose dolphins (Tursiops truncatus) were analysed to determine whether signature whistles are produced by wild undisturbed dolphins, and how whistle production varies with activity and group size. The study animals were part of a resident community of bottlenose dolphins near Sarasota, Florida, USA. This community of dolphins provides a unique opportunity for the study of signature-whistle production, since most animals have been recorded during capture-release events since 1975. Three mother-calf pairs and their associates were recorded for a total of 141.25 h between May and August of 1994 and 1995. Whistles of undisturbed dolphins were compared with those recorded from the same individuals during capture-release events. Whistles were conservatively classified into one of four categories: signature, probable signature, upsweep or other. For statistical analyses, signature and probable signature whistles were combined into a 'signature' category; upsweep and other whistles were combined into a 'non-signature' category. Both 'signature' and 'non-signature' whistle frequencies significantly increased as group size increased. There were significant differences in whistle frequencies across activity types: both 'signature' and 'non-signature' whistles were most likely to occur during socializing and least likely to occur during travelling. There were no significant interactions between group size and activity type. Signature and probable signature whistles made up ca. 52% of all whistles produced by these free-ranging bottlenose dolphins.     

Food-related bray calls in wild bottlenose dolphins (Tursiops truncatus)

Because cetaceans are difficult to study in the wild, little is known about how they use their sounds in their natural environment. Only the recent development of passive acoustic localization systems has enabled observations of the communication behaviour of individuals for correlation with their surface behaviour. Using such a system, I show that bottlenose dolphins in the Moray Firth, Scotland, produce low-frequency bray calls which are clearly correlated with feeding on salmonids. The production of these calls is followed by fast approaches by conspecifics in the area. In animals which use sound as a foraging tool, it is difficult to distinguish between food calls which have evolved because of their role in attracting conspecifics, and food manipulation or searching calls which may attract conspecifics as a by-product. However, the low-frequency structure of the bottlenose dolphin bray suggests that it evolved because of a role in manipulating prey rather than in attracting conspecifics. This conclusion suggests that dolphins exploit the perceptual systems of their prey to facilitate capture.     

Lateralized behavior in two captive bottlenose dolphins (Tursiops truncatus)

The study of behavioral laterality in humans and nonhumans can contribute to our understanding of brain evolution and functional similarities across species. Few studies have focused on cetaceans. This report exams lateralized behaviors in two captive bottlenose dolphins (Tursiops truncatus). Observations were made by videotaping through a 90 × 150 cm underwater one-way Plexiglass mirror. Directional bias in swimming, “barrel-roll” maneuvers, and circular head movements was assessed for each subject. There was a strong clockwise bias in swimming direction and direction of “barrel-rolls,” but not circular head movements. The clockwise bias in swimming direction and “barrel-roll” maneuvers may be consistent with a rightward turning bias. Zoo Biol 16:173–177, 1997. © 1997 Wiley-Liss, Inc.     

Fat distribution in the skin of bottlenose dolphins (Tursiops truncatus and Tursiops gilli)

Frozen sections stained with Oil-red-O and semithin (0.5 μm) plastic sections stained with toluidine blue revealed an abundance of fat globules of various sizes in all strata of the epidermis of bottlenose dolphins (Tursiops truncatus and T. gilli). The fat was rather evenly distributed but sometimes appeared as circumscribed areas of heavier concentration involving hundreds of cells (as seen in a single plane). Occasionally, there were smaller groups of epidermal cells heavily loaded with lipid. The dermis presented a unique phenomenon in the presence of abundant extracellular fat distributed among the collagen bundles as droplets of various sizes or as larger, irregularly shaped lipid particles that seemed to conform to the spaces between collagen bundles. These lipid particles were sometimes seen to be closely applied to the dermal surface of the stratum basale. Equally unusual was the presence of lipid particles of various sizes and shapes in the lumen of some of the vessels of the dermal papillae. Granular cells resembling mast cells were commonly seen in the papillary dermis and some were closely associated with lipid particles. Blood vessels of the reticular dermis tended to have collections of lipid droplets in the loose connective tissue often found adjacent to the tunica adventitia. It is postulated that the extracellular dermal lipids (probably mainly triglycerides) are broken down to free fatty acids that diffuse into the basal layer of the epidermis and are there resynthesized into triglycerides. Possible uses for the epidermal lipids are discussed.     

Isolation of Toxoplasma gondii from bottlenose dolphins (Tursiops truncatus)

Toxoplasma gondii infection in marine mammals is intriguing and indicative of contamination of the ocean environment and coastal waters with oocysts. In previous serological surveys, >90% of bottlenose dolphins (Tursiops truncatus) from the coasts of Florida, South Carolina, and California had antibodies to T. gondii by the modified agglutination test (MAT). In the present study, attempts were made to isolate T. gondii from dead T. truncatus. During 2005, 2006, and 2007, serum or blood clot, and tissues (brain, heart, skeletal muscle) of 52 T. truncatus stranded on the coasts of South Carolina were tested for T. gondii. Antibodies to T. gondii (MAT 1:25 or higher) were found in 26 (53%) of 49 dolphins; serum was not available from 3 animals. Tissues (heart, muscle, and sometimes brain) of 32 dolphins (26 seropositive, 3 seronegative, and 3 without accompanying sera) were bioassayed for T. gondii in mice, or cats, or both. Tissues of the recipient mice were examined for T. gondii stages. Feces of recipient cats were examined for shedding of T. gondii oocysts, but none excreted oocysts. Toxoplasma gondii was isolated from hearts of the 3 dolphins (2 with MAT titers of 1:200, and 1 without accompanied serum) by bioassay in mice. Genotyping of these 3 T. gondii isolates (designated TgDoUs1-3) with the use of 10 PCR-RFLP markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico) revealed 2 genotypes. Two of the 3 isolates have Type II alleles at all loci and belong to the clonal Type II lineage. One isolate has a unique genotype. This is the first report of isolation of viable T. gondii from T. truncatus.     

Fatal Asphyxiation in Bottlenose Dolphins (Tursiops truncatus) from the Indian River Lagoon

Multiple single case reports of asphyxiation in dolphins caused by fish lodged in the esophagus exist. However, the significance of this cause of mortality in a single population has not been documented. We performed a retrospective evaluation of pathology records from stranded bottlenose dolphins (Tursiops truncatus) from the Indian River Lagoon to evaluate the impact of this cause of death on this population. From 1997 to 2011, asphyxiation due to choking was identified as the cause of death in 14 of 350 cases (4%). Sampling of an unrelated but adjacent population over this same period yielded 186 necropsy cases of bottlenose dolphins with no cases of asphyxiation. Asphyxiated animals presented with a fish lodged in the cranial esophagus associated with a dislocated and obstructed or compressed larynx. There was no clear sex predilection. Affected animals included 12 adults and two juveniles. The fish species involved included sheepshead, black chin tilapia and striped mojarra. In five cases, recreational fishing gear was also present. Cetacean choking is related to selection of prey fish species with strong dorsal spines and may be secondarily associated with fish attached to fishing gear. Prey abundance and dolphin behavior may influence these selections. Environmental alterations leading to changes in prey availability or increased interactions with fishing gear may change the significance of fatal choking in dolphin populations.     

Precocial development of axial locomotor muscle in bottlenose dolphins (Tursiops truncatus)

At birth, the locomotor muscles of precocial, terrestrial mammals are similar to those of adults in both mass, as a percent of total body mass, and fiber-type composition. It is hypothesized that bottlenose dolphins (Tursiops truncatus), marine mammals that swim from the instant of birth, will also exhibit precocial development of locomotor muscles. Body mass data from neonatal and adult dolphins are used to calculate Grand's (1992) Neural and Muscular Indices of Development. Using these indices, the bottlenose dolphin is a Condition "3.5" neonate, where Condition 4 is the documented extreme of precocial development in terrestrial mammals. Moreover, myosin ATPase (alkaline preincubation) analyses of the epaxial locomotor m. extensor caudae lateralis show that neonatal dolphins have fiber-type profiles very similar to those of adults. Thus, based on mass and myosin ATPase activity, muscle development in dolphins is precocial. However, succinic dehydrogenase and Nile red histochemistry demonstrate that neonatal dolphin muscle has mitochondrial and lipid distributions different from those found in adults. These data suggest that neonates have a lower aerobic capacity than adults. Dolphin neonates may compensate for an apparent lack of aerobic stamina in two ways: 1) by being positively buoyant, with a relatively increased investment of their total body mass in blubber, and 2) by "free-riding" off their mothers. This study investigates quantitatively the development of a dolphin locomotor muscle and offers suggestions about adaptations required for a completely aquatic existence.     

Captive Bottlenose Dolphins (Tursiops truncatus) Spontaneously Using Water Flow to Manipulate Objects

Several terrestrial animals and delphinids manipulate objects in a tactile manner, using parts of their bodies, such as their mouths or hands. In this paper, we report that bottlenose dolphins (Tursiops truncatus) manipulate objects not by direct bodily contact, but by spontaneous water flow. Three of four dolphins at Suma Aqualife Park performed object manipulation with food. The typical sequence of object manipulation consisted of a three step procedure. First, the dolphins released the object from the sides of their mouths while assuming a head-down posture near the floor. They then manipulated the object around their mouths and caught it. Finally, they ceased to engage in their head-down posture and started to swim. When the dolphins moved the object, they used the water current in the pool or moved their head. These results showed that dolphins manipulate objects using movements that do not directly involve contact between a body part and the object. In the event the dolphins dropped the object on the floor, they lifted it by making water flow in one of three methods: opening and closing their mouths repeatedly, moving their heads lengthwise, or making circular head motions. This result suggests that bottlenose dolphins spontaneously change their environment to manipulate objects. The reason why aquatic animals like dolphins do object manipulation by changing their environment but terrestrial animals do not may be that the viscosity of the aquatic environment is much higher than it is in terrestrial environments. This is the first report thus far of any non-human mammal engaging in object manipulation using several methods to change their environment.     

Southernmost records of bottlenose dolphins, Tursiops truncatus

Bottlenose dolphins (Tursiops spp.) are cosmopolitan animals widely distributed in waters of both hemispheres. The taxonomy of Tursiops has long been controversial, with over 20 specific names being published, and subspecies and inshore/offshore forms being proposed. In the southwestern South Atlantic, subspecies T. truncatus truncatus and T. truncatus gephyreus were proposed for specimens along the coasts of Brazil, Uruguay, and Argentina. Sightings of bottlenose dolphins are common along the coast of Argentina as far south as the Province of Chubut (ca. 46°S). Here, we summarize and discuss the southernmost records of bottlenose dolphins. We cannot make inferences about the species or subspecies to which these animals belong given the small number of specimens. Future studies of external measurements, pigmentation, DNA, and isotopes from both sides of the continent should help clarify the situation off southern South America. Furthermore, research is needed to explore a possible link between an effect of general global warming and the Tursiops specimens found this far south. The sighting and specimens described here, at 53°S–nearly 55°S, are the southernmost records for the genus and extend the range of the species in the southern South Atlantic.     

Spatial segregation and interspecific killing of common dolphins (Delphinus delphis) by bottlenose dolphins (Tursiops truncatus)

acta ethologica - We described the spatial segregation of two species of cetaceans, the common dolphin and the bottlenose dolphin. We also document the first direct observation of interspecific...     

Postural role of lateral axial muscles in developing bottlenose dolphins (Tursiops truncatus)

Foetal dolphins (Tursiops truncatus) are bent ventrolaterally, such that the tailflukes and lower jaw are juxtaposed. The lateral flexibility required en utero may compromise the efficiency of the dorsoventral oscillations required of the swimming neonate. The m. intertransversarius caudae dorsalis (IT) is the most laterally placed epaxial muscle. Bilateral contractions of the IT could limit lateral deformations of the flexible tailstock of the early neonate. We test the hypothesis that the IT is functioning as a postural muscle in neonates by examining its morphological, histological and biochemical properties. The neonatal IT has a relatively large cross-sectional area and bending moment, as well as a large proportion of slow-twitch fibres and elevated myoglobin concentrations. Our results demonstrate that the IT is functionally capable of performing this specific postural function in neonatal dolphins. In later life-history stages, when postural control is no longer needed, the IT serves to fine-tune the position of the tailstock during locomotion. The changing function of the adult IT is concomitant with changes in morphology and biochemistry, and most notably, with an increase in the proportion of fast-twitch fibres. We suggest that these changes reflect strong selective pressure to improve locomotor abilities by limiting lateral deformations during this critical life-history stage.     

Incidental capture of bottlenose dolphins (Tursiops truncatus) in shark nets: an assessment of some possible causes

A minimum of 250 bottlenose dolphins, Tursiops truncatus , was caught in anti-shark nets off Natal, South Africa between January 1980 and December 1988. Males and females constituted 45.4% and 54.6% of the catch, respectively. Sexually mature males and females constituted 18.2% and 24% of the total catch, respectively. Of the latter, 65% were lactating and a further 10.3% were pregnant. Adolescents, three or more years old but not mature, contributed 12.4% of the catch. Calves, two growth layer groups (GLGs) or less in the dentine, constituted the greatest portion of the catch, almost 43%, and of these almost 70% were either weaned or weaning at capture. Analyses of biological, environmental and physiographic data for each capture suggest a number of reasons for the catch of bottlenose dolphins. The distribution of catches was random but catch rates were proportional to the number of nets. The stomachs of most dolphins were almost full at the time of capture, suggesting that enmeshing occurred either during or subsequent to feeding. Most captures were of single animals, but lactating females with calves constituted more than 25% of catches. The direction of the prevailing current was significantly related to captures. These data are examined in relation to existing knowledge of bottlenose dolphins in this area and suggest that capture may be a consequence of prey abundance at and around the nets, dolphin capture occurring either during or directly after feeding. Possible methods of reducing captures are proposed.     

Identification and Characteristics of Signature Whistles in Wild Bottlenose Dolphins (Tursiops truncatus) from Namibia

A signature whistle type is a learned, individually distinctive whistle type in a dolphin''s acoustic repertoire that broadcasts the identity of the whistle owner. The acquisition and use of signature whistles indicates complex cognitive functioning that requires wider investigation in wild dolphin populations. Here we identify signature whistle types from a population of approximately 100 wild common bottlenose dolphins (Tursiops truncatus) inhabiting Walvis Bay, and describe signature whistle occurrence, acoustic parameters and temporal production. A catalogue of 43 repeatedly emitted whistle types (REWTs) was generated by analysing 79 hrs of acoustic recordings. From this, 28 signature whistle types were identified using a method based on the temporal patterns in whistle sequences. A visual classification task conducted by 5 naïve judges showed high levels of agreement in classification of whistles (Fleiss-Kappa statistic, κ = 0.848, Z = 55.3, P<0.001) and supported our categorisation. Signature whistle structure remained stable over time and location, with most types (82%) recorded in 2 or more years, and 4 identified at Walvis Bay and a second field site approximately 450 km away. Whistle acoustic parameters were consistent with those of signature whistles documented in Sarasota Bay (Florida, USA). We provide evidence of possible two-voice signature whistle production by a common bottlenose dolphin. Although signature whistle types have potential use as a marker for studying individual habitat use, we only identified approximately 28% of those from the Walvis Bay population, despite considerable recording effort. We found that signature whistle type diversity was higher in larger dolphin groups and groups with calves present. This is the first study describing signature whistles in a wild free-ranging T. truncatus population inhabiting African waters and it provides a baseline on which more in depth behavioural studies can be based.     

Patterns of growth in wild bottlenose dolphins, Tursiops truncatus

The growth of bottlenose dolphins is described from observations made during a capture release programme that has operated in coastal waters of the eastern Gulf of Mexico from 1970 to the present. Measurements of standard length, girth and body mass were recorded from 47 female and 49 male dolphins, some captured as many as nine times. Ages were known from approximate birth dates or estimated from counts of dentinal growth layers. In all three measurements. females grew at a faster initial rate than males, but reached asymptotic size at an earlier age. This extended period of growth in males resulted in significant sexual dimorphism in length, girth and mass at physical maturity. The growth of both sexes was well described by three-parameter Gompertz models using either cross-sectional data or a mixture of longitudinal and cross-sectional data. There was considerable variation in size-at-age for both sexes in all year classes. Residuals of size measurements were used to derive measures of relative size for individual dolphins; most dolphins demonstrated little ontogenetic change in relative size. Body mass was adequately predicted by multiple regression equations that incorporated both length and girth as independent variables.     

High gene flow in oceanic bottlenose dolphins (Tursiops truncatus) of the North Atlantic

Despite the openness of the oceanic environment, limited dispersal and tight social structure often induce genetic structuring in marine organisms, even in large animals such as cetaceans. In the bottlenose dolphin, mitochondrial and nuclear DNA analyses have revealed the existence of genetic differentiation between pelagic (or offshore) and coastal (or nearshore) ecotypes in the western North Atlantic, as well as between coastal populations. Because previous studies concentrated on continental margins, we analysed the population structure of bottlenose dolphins in two of the most isolated archipelagos of the North Atlantic: the Azores and Madeira. We analysed 112 samples collected on live animals in the two archipelagos, and nine samples collected on stranded animals in Madeira and mainland Portugal. Genetic analyses consisted in molecular sexing, sequencing of part of the mitochondrial hyper-variable region, and screening of ten microsatellite loci. We predicted that: (1) there is at least one pelagic and one or more coastal populations in each archipelago; (2) populations are differentiated between and possibly within archipelagos. Contrary to these predictions, results indicated a lack of population structure in the study area. In addition, comparison with published sequences revealed that the samples from the Azores and Madeira were not significantly differentiated from samples of the pelagic population of the western North Atlantic. Thus, bottlenose dolphins occurring in the pelagic waters of the North Atlantic belong to a large oceanic population, which should be regarded as a single conservation unit. Unlike what is known for coastal populations, oceanic bottlenose dolphins are able to maintain high levels of gene flow.     

Genetic isolation of a now extinct population of bottlenose dolphins (Tursiops truncatus)

A number of dolphin species, though highly mobile, show genetic structure among parapatric and sometimes sympatric populations. However, little is known about the temporal patterns of population structure for these species. Here, we apply Bayesian inference and data from ancient DNA to assess the structure and dynamics of bottlenose dolphin (Tursiops truncatus) populations in the coastal waters of the UK. We show that regional population structure in UK waters is consistent with earlier studies suggesting local habitat dependence for this species in the Mediterranean Sea and North Atlantic. One genetically differentiated UK population went extinct at least 100 years ago and has not been replaced. The data indicate that this was a local extinction, and not a case of historical range shift or contraction. One possible interpretation is a declining metapopulation and conservation need for this species in the UK.     

The Orexin System in the Enteric Nervous System of the Bottlenose Dolphin (Tursiops truncatus)

This study provides a general approach to the presence and possible role of orexins and their receptors in the gut (three gastric chambers and intestine) of confined environment bottlenose dolphin. The expression of prepro-orexin, orexin A and B and orexin 1 and 2 receptors were investigated by single immunostaining and western blot analysis. The co-localization of vasoactive intestinal peptide and orexin 1 receptor in the enteric nervous system was examined by double immunostaining. Also, orexin A concentration were measured in plasma samples to assess the possible diurnal variation of the plasma level of peptide in this species. Our results showed that the orexin system is widely distributed in bottlenose dolphin enteric nervous system of the all gastrointestinal tract examined. They are very peculiar and partially differs from that of terrestrial mammals. Orexin peptides and prepro-orexin were expressed in the main stomach, pyloric stomach and proximal intestine; while orexin receptors were expressed in the all examined tracts, with the exception of main stomach where found no evidence of orexin 2 receptor. Co-localization of vasoactive intestinal peptide and orexin 1 receptor were more evident in the pyloric stomach and proximal intestine. These data could suggest a possible role of orexin system on the contractility of bottlenose dolphin gastrointestinal districts. Finally, in agreement with several reports, bottlenose dolphin orexin A plasma level was higher in the morning during fasting. Our results emphasize some common features between bottlenose dolphin and terrestrial mammals. Certainly, further functional investigations may help to better explain the role of the orexin system in the energy balance of bottlenose dolphin and the complex interaction between feeding and digestive physiology.