Lymphangiomyomatosis in the Atlantic bottlenose dolphin (Tursiops truncatus).

The first case of lymphangiomyomatosis in a marine mammal is reported from a stranded male Atlantic bottlenose dolphin (Tursiops truncatus). This progressive proliferative disease involved the lungs and the mediastinal and probably mesenteric lymph nodes. An extraordinarily low level of testosterone may have been pathologically significant since all reported cases of this disease in humans occur in females.     

Vertebral osteomyelitis and suspected diskospondylitis in an Atlantic bottlenose dolphin (Tursiops truncatus)

A 21-yr-old male Atlantic bottlenose dolphin (Tursiops truncatus) was performing at an aquatic park when it developed a soft tissue swelling anterior to the flukes. Subsequent radiographic evaluation revealed the animal to have vertebral osteomyelitis and suspected diskospondylitis. The case was successfully managed with long-term antibiotic therapy.     

Harderian gland ultrastructure of the black sea bottlenose dolphin (Tursiops truncatus ponticus)

Examination of the Harderian gland structure of the Black Sea bottlenose dolphin, Tursiops truncatus ponticus, at macroscopic, microscopic, and electron microscopic levels shows significant sexual dimorphism. The epithelial cells of male and female glands are different cell types, capable of producing chemically different products. Secretory cells in both sexes contain secretion granules that produce a secretion consisting mainly of proteins and carbohydrates, but thought to be sex-specific in composition. The female glands also contain lipid secretion granules. It is suggested that in the bottlenose dolphin the Harderian gland functions to produce sexually distinct pheromones and may have other physiological activities, e.g., participating in local immunological or endocrine-related reactions. © 1994 Wiley-Liss, Inc.     

Assessment of increased serum aminotransferases in a managed Atlantic bottlenose dolphin (Tursiops truncatus) population

Nonspecific chronic hepatitis and increased activities of serum aminotransferases have been reported in cetaceans (dolphins, porpoises, and whales). We identified bottlenose dolphins in our current population with episodic increases in serum aminotransferases, specifically alanine aminotransferase (ALT) and aspartate aminotransferase (AST), and we hypothesized that hematologic and serum biochemical changes in these animals may provide clues as to potential causes of liver disease in cetaceans. A retrospective case-control study involving 1,288 blood samples collected during 1998-2006 from 18 dolphins (six cases and 12 age- and sex-matched healthy controls) was conducted to compare eosinophil and platelet counts; and serum proteins, albumin, globulins, bilirubin, gamma glutamyltransferase (GGT), cholesterol, triglycerides, glucose, iron, and erythrocyte sedimentation rates. Bottlenose dolphins with increased ALT and AST activities were more likely to have higher serum globulins, bilirubin, GGT, iron, glucose, triglycerides, and cholesterol levels, greater erythrocyte sedimentation rates, and lower platelet counts compared to healthy controls. Our findings suggest that dolphins with chronic increases in aminotransferases may have a chronic hepatitis involving iron overload with similar etiologies and pathophysiology compared to terrestrial mammals. Areas for future research include predisposing metabolic risk factors; associations between iron overload and a diabetes-like condition; and a potential overlap syndrome involving autoimmune responses that may or may not be associated with viral infection.     

The Immunoglobulin G Heavy Chain (IGHG) genes of the Atlantic bottlenose dolphin, Tursiops truncatus

Dolphin Immunoglobulin G Heavy Chain (IGHG) sequences were obtained by PCR amplification of cDNA from peripheral blood leukocytes using degenerate primers. Analysis of full-length sequences indicated the presence of two expressed isotypes, IGHG1 and IGHG2 that differ mainly in the hinge region of the molecule. Genomic Southern blot analysis indicated that the IGHG1 and IGHG2 genes are most likely present in single copies. The inferred amino acid sequences show greatest similarity between the dolphin and other closely related artiodactyl species. The genetic structure of the IGHG genes were deduced through genomic PCR and revealed that the hinge regions of both IGHG1 and IGHG2 are encoded by a single exon. The transmembrane region of the dolphin IGHG chain shows similarity to the transmembrane region of other mammalian IGHG chains with a canonical CART motif. This is in contrast to the unusual Ser to Gly substitution previously found in the dolphin IGHM transmembrane region, and the functional significance of this variation for B cell antigen-receptor dimer activation remains unknown.     

Struvite calculus in the vagina of a bottlenose dolphin (Tursiops truncatus)

On 27 January 2000, a struvite calculus was observed in the vagina during necropsy of a 138-cm-long female bottlenose dolphin (Tursiops truncatus) collected from the Stono River, South Carolina (USA). Vaginal calculi have been reported in other species of cetaceans but not in bottlenose dolphins. Urinary tract infection might have been an underlying cause of the calculus. While urinary tract inflammation was not detected by light microscopic evaluation of sections of the urinary tract, it is conceivable that sufficient time had lapsed following voiding of the calculus through the urethra for urinary tract infection to have resolved. To further define the prevalence and significance of urolithiasis, prosectors of dead stranded marine mammals are encouraged to closely observe their urinary and genital tracts for calculi and to submit them for quantitative analysis.     

Effect of cryopreservation methods and precryopreservation storage on bottlenose dolphin (Tursiops truncatus) spermatozoa

Research was conducted to develop an effective method for cryopreserving bottlenose dolphin (Tursiops truncatus) semen processed immediately after collection or after 24-h liquid storage. In each of two experiments, four ejaculates were collected from three males. In experiment 1, three cryopreservation methods (CM1, CM2, and CM3), two straw sizes (0.25 and 0.5 ml), and three thawing rates (slow, medium, and fast) were evaluated. Evaluations were conducted at collection, prefreeze, and 0-, 3-, and 6-h postthaw. A sperm motility index (SMI; total motility [TM] x % progressive motility [PPM] x kinetic rating [KR, scale of 0-5]) was calculated and expressed as a percentage MI of the initial ejaculate. For all ejaculates, initial TM and PPM were greater than 85%, and KR was five. At 0-h postthaw, differences in SMI among cryopreservation methods and thaw rates were observed (P < 0.05), but no effect of straw size was observed. In experiment 2, ejaculates were divided into four aliquots for dilution (1:1) and storage at 4 degrees C with a skim milk- glucose or a N-tris(hydroxymethyl)methyl-2-aminoethane sulfonic acid (TES)-TRIS egg yolk solution and at 21 degrees C with a Hepes-Tyrode balanced salt solution (containing bovine albumin and HEPES) (TALP) medium or no dilution. After 24 h, samples were frozen and thawed (CM3, 0.5-ml straws, fast thawing rate) at 20 x 10(6) spermatozoa ml(-1) (low concentration) or at 100 x 10(6) spermatozoa ml(-1) (standard concentration). The SMI at 0-h postthaw was higher for samples stored at 4 degrees C than for samples stored at 21 degrees C (P < 0.001), and at 6-h postthaw, the SMI was higher for samples frozen at the standard concentration than for samples frozen at the low concentration (P < 0.05). For both experiments, acrosome integrity was similar across treatments. In summary, a semen cryopreservation protocol applied to fresh or liquid-stored semen maintained high levels of initial ejaculate sperm characteristics.     

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.     

长江江豚和瓶鼻海豚的血清转铁蛋白

血清转铁蛋白(Serum transferrin,Tf)是广泛存在于血清中的β球蛋白,其主要功能是将铁离子以安全的形式(Fe3 )在机体的吸收、储存和利用部位之间进行传递,为细胞生长提供铁(Fe3 )(Welch,1990);同时具有保护细胞、促进细胞分化和生长、抑菌和杀菌的作用(Paezetal.,2002;Liuet al     

Occurrence of a unique sialyl tetrasaccharide in colostrum of a bottlenose dolphin (Tursiops truncatus)

Crude oligosaccharides were recovered from bottlenose dolphin (Tursiops truncatus) colostrum after chloroform/methanol extraction of lipids and protein precipitation, and purified using gel filtration, anion exchange chromatography and high performance liquid chromatography (HPLC). Their chemical structures characterized by NMR spectroscopy were as follows: GalNAc(beta1-4)[Neu5Ac(alpha2-3)]Gal(beta1-4)Glc, Neu5Ac(alpha2-3)Gal(beta1-4)Glc, Neu5Ac(alpha2-6)Gal(beta1-4)Glc and Gal(alpha1-4)Gal(beta1-4)Glc. The monosialyltetrasaccharide and neutral trisaccharide have not previously been found as free forms in any natural sources including milk or colostrum, although these structures have been found in the carbohydrate units of glycosphingolipids GM2 and Gb3.     

Importance of spontaneous micronucleated erythrocytes in bottlenose dolphin (Tursiops truncatus) to marine toxicology studies

The objective of the work was to characterize the presence of spontaneous micronucleated erythrocytes (MNES) from peripheral blood of bottlenose dolphins (Tursiops truncatus) to evaluate the possibility to use this species as potential bioindicator of genotoxic compounds. Forty-eight blood samples from 12 bottlenose dolphins were obtain from three Mexican dolphinariums, and from 10 dolphins was possible to obtain more than one sample at different sampling times. Smears were processed and observed with an epifluorescence microscope. The average of MNES and polychromatic erythrocytes (PCE) from the 48 samples was 24.3 +/- 6.1 MNES/10,000 total erythrocytes (TE), and 9.1 +/- 5.5 PCE/1,000 TE. MNES and PCE number did not show differences between gender and age. No variations in the MNES values of the bottlenose dolphins that were sampled more than one occasion were found. Comparisons among dolphinariums revealed differences in MNES frequency, with the highest significant frequency observed in dolphins from dolphinarium "A" (26.0 +/- 5.9 MNES/10,000 TE) than dolphinarium "B" (19.5 +/- 3.1 MNES/10,000 TE) (p < 0.05) and dolphinarium "C" (18.6 +/- 3.5 MNES/10,000 TE) (p < 0.007). The presence of MNES and PCE in the bottlenose dolphin may provide a useful marine mammal model to detect DNA damage by means of micronuclei test in peripheral blood erythrocytes to evaluate genotoxicity and cytotoxicity expositions.     

Molecular cloning and histological localization of LH-like substances in a bottlenose dolphin (Tursiops truncatus) placenta

All mammals exhibit pituitary-specific expression of LH and FSH, whereas placental expression of gonadotropins has been reported only in primates and equids. Some cetaceans, such as dolphins, have a long gestational period and a sexual cycle of about 27 days almost comparable with that of humans. Histologically, dolphins have an epitheliochorial placentae that resembles placentas of Perissodactyla including horses. In the present study, we cloned cDNAs encoding gonadotropins and observed their immunohistochemical localization in the placenta of bottlenose dolphin. The cDNAs obtained encoded 120 amino acids for the alpha-subunit (including 96 amino acids of mature proteins), and 141 amino acids for the beta-subunit (including 121 amino acids of mature proteins). The sequence of the alpha-subunit was similar to that in the pig (Artiodactyla) pituitary glycoprotein hormone [96.7% homology at amino acids (aa) level], and the sequence of the beta-subunit was similar to that of luteinizing hormone (LH) in the pig [94.3% homology at aa level] and white rhinoceros (Perissodactyla) [93.3% homology at aa level]. Of interest, dolphin LHbeta lacks carboxyl-terminal-peptides (CTP). This fact suggests that CTP are not essential for placental expression of gonadotropin in dolphins. Immunohistochemical observations employing anti-ovine LHbeta antibody revealed positive staining in the villositycal tissue. Our observations suggest placental expression of gonadotropin homologues in cetaceans and possible evolutionary conservation of placentae-derived hormonal control of ovarian functions during pregnancy.     

Progress report: Results of a long-range captive breeding program for the bottlenose dolphin,Tursiops truncatus and Tursiops truncatus gilli

There have been 36 bottlenose dolphins born in breeding colonies at Sea World, California, and Sea World, Florida, from 1978–1985. Significant features of this successful reproductive program are construction of a compatible breeding colony, early hormonal detection of pregnancy, pre- and postnatal association of inexperienced mothers with experienced females and their calves, and minimized handling of the mothers and calves until the calves are over 1 year of age. Inclusion in the breeding colonies of males old enough to be effective breeders is stressed. Females in the colonies have successfully bred from 8–9 years of age to 23–24 years of age. Calving intervals in the colonies vary from 2 to over 3 years. Calves are born all year round, with some peaking in calving activity in the spring and fall months. Serum progesterone levels greater than 6,000 pg/ml, maintained over a 4–6-week period, are considered indicative of pregnancy. Progesterone levels vary from less than 10,000 to over 50,000 pg/ml during gestation, averaging 25,000 pg/ml. The need for further study of variation in the pattern of progesterone levels during pregnancy is emphasized. A gestation period for Tursiops of 11.5 to 12 months is consistent with our cumulative progesterone data.     

Thirty year retrospective evaluation of pneumonia in a bottlenose dolphin Tursiops truncatus population

Pneumonia is one of the most common causes of morbidity in bottlenose dolphins Tursiops truncatus. To better understand associations of pneumonia with demographics, microbiology, pathology, and histopathology, a retrospective study on 42 dolphins from the US Navy Marine Mammal Program dolphin population was conducted (1980 to 2010). A total of 21 (50%) of the dolphins evaluated had pneumonia confirmed by histopathology. Bacterial and fungal pneumonia was present in 42.9 and 28.6% of cases (9 and 6 cases), respectively, with Staphylococcus aureus as the most common confirmed pathogen (4 cases, 19%). Other pathogens identified as the cause of pneumonia were Cryptococcus neoformans, Erysipelothrix rhusiopathiae, Histoplasma capsulatum, parainfluenza virus, Proteus species, Pseudomonas aeruginosa, and Streptococcus zooepidemicus. Neither sex nor age was a predictor of pneumonia. While many of the infections involved disseminated disease, lungs were consistently the most severely affected organs. The present study demonstrates the high susceptibility of dolphins to respiratory infections. Areas that warrant further investigation include eosinophilic pneumonia, chronic infections, co-infections, and metabolic or iron-storage diseases. There is a continuing need to improve the early diagnosis and effective treatment of pneumonia in dolphins.     

First case of erysipelas in a free-ranging bottlenose dolphin (Tursiops truncatus) stranded in the Mediterranean Sea

An adult female bottlenose dolphin (Tursiops truncatus) stranded alive and subsequently died several minutes later on the Mediterranean coast of Spain on 14 July 2010. Clinical examination revealed foam through the blowhole and rales upon lung auscultation. On venipuncture, the blood was abnormally dense and dark. Hematological and biochemical abnormalities included dehydration, leukocytosis (48 600 leukocytes microl(-1)) characterized by neutrophilia (48 200 neutrophils microl(-1)), and elevated bilirubin (4.38 mg dl(-1)), alanine aminotransferase (382.3 U l(-1)), aspartate aminotransferase (1449.3 U l(-1)), lactate dehydrogenase (1631.3 U l(-1)), and creatine kinase (404.7 U l(-1)). The most relevant findings of the gross examination were rhomboid-shaped skin lesions, stable froth in the trachea, pulmonary congestion, abnormally thick and rough pleura with adhesions, edematous and congestive superficial cervical and tracheobronchial lymph nodes, red-tinged urine, and severe brain congestion. Histopathology of the kidney, lung, skin, and brain revealed multisystemic intravascular bacterial emboli. Samples of skin, brain, and lung were cultured on Columbia blood agar under both aerobic and anaerobic conditions, and pure and heavy bacterial cultures were obtained from skin and brain samples. The microorganism isolated was Gram-positive, catalase-negative, facultatively anaerobic, and rod-shaped. The isolates were identified as Erysipelothrix rhusiopathiae by the API Coryne biochemical system. Based on the gross and microscopic findings, a diagnosis of acute E. rhusiopathiae septicemia was made. To the best of our knowledge, this is the first report of E. rhusiopathiae septicemia in a free-ranging bottlenose dolphin.     

Temperature regulation of the testes of the bottlenose dolphin (Tursiops truncatus): evidence from colonic temperatures

Dolphins possess a countercurrent heat exchanger that functions to cool their intra-abdominal testes. spermatic arteries in the posterior abdomen are juxtaposed to veins returning cooled blood from the surfaces of the dorsal fin and flukes. A rectal probe housing a linear array of five copper-constantan thermocouples was designed to measure colonic temperatures simultaneously at positions anterior to, within, and posterior to the region of the colon flanked by the countercurrent heat exchanger. Colonic temperatures adjacent to the countercurrent heat exchanger were maximally 1.3°C cooler than temperatures measured outside this region. Temporary heating and cooling of the dorsal fin and flukes affected temperatures at the countercurrent heat exchanger, but had little or no effect on temperatures posterior to its position. These measurements support the hypothesis that cooled blood is introduced into the deep abdominal cavity and functions specifically to regulate the temperature of arterial blood flow to the dolphin testes.