Effect of bovine lactoferrin on the immune responses of captive bottlenosed dolphins (Tursiops truncatus) being transported over long distances

Bovine lactoferrin was administered orally, in feed, to six bottlenosed dolphins (Tursiops truncatus) before they were transported for approximately six hours; their stress responses were compared with those of five untreated dolphins. During the journey the dolphins had an increased plasma concentration of cortisol, and lymphopenia, eosinopenia and mild neutrophilia, indicating a stress response. The administration of lactoferrin did not affect the function of the dolphins' polymorphonuclear leucocytes, but affected their leucogram by maintaining the number of circulating eosinophils.     

Altrenogest and progesterone therapy during pregnancy in bottlenose dolphins (Tursiops truncatus) with progesterone insufficiency

Progesterone production is essential for growth and development of the conceptus during pregnancy. Abnormal development of the corpus luteum (CL) after conception can result in early embryonic loss or fetal abortion. Routine monitoring of bottlenose dolphin (Tursiops truncatus) pregnancy after artificial insemination or natural conception with ultrasonography and serum progesterone determination has allowed for the establishment of expected fetal growth rates and hormone concentrations. Using these monitoring techniques, we revealed four pregnant dolphins (12-24 yr old) with abnormally low progesterone production indicative of luteal insufficiency. Once diagnosed, animals were placed on altrenogest (0.044-0.088 mg/kg once daily) alone or with oral progesterone (50-200 mg twice daily). Doses of hormone were increased or decreased in each animal based on how fetal skull biparietal and thoracic growth rates compared with published normal values. Hormones were withdrawn starting from day 358 of gestation in animals 1 and 2, with labor occurring 6 and 7 days after withdrawal and at 376 and 373 days of gestation, respectively. Both deliveries were dystocic, with each calf requiring manual extraction and fetotomy for calf 1. The fetuses in animals 3 and 4 died at 348 and 390 days of gestation, respectively. Induction of labor was attempted in both animals, after fetal death, by using a combination of rapid progesterone withdrawal and steroid and prostaglandin F2alpha administration. The calf of animal 4 had to be removed with manual cervical dilation and fetotomy All adult females survived the procedures. These data provide the first in vivo evidence that the CL is the primary source of progesterone throughout pregnancy in the bottlenose dolphin. Until further characterization of hormones required during pregnancy and at parturition has been accomplished, the exogenous progestagen supplementation protocol described here cannot be recommended for treatment of progesterone insufficiency in bottlenose dolphins.     

Prediction of gestational age by transrectal ultrasonographic measurements in the mouflon (Ovis gmelini musimon).

Serial transrectal ultrasonographies were performed measuring different structures of mouflon (Ovis gmelini musimon) fetuses to establish their gestational age. Data were obtained from 14 mouflon ewes that became pregnant after estrous synchronization. From day 25 of gestation onward, the crown-rump length and the trunk diameter of the embryo were estimated. Later, when the fetal organs and structures could be distinguished, several measurements were done in the skull (occipito-snout length and biparietal and orbit diameters), the trunk (thoracic diameter and width of vertebrae), and the abdominal cavity (stomach and kidney longitudinal diameters); femur length was also determined. All fetal measurements were correlated with gestational age (P < 0.001), but the parameters showing the highest correlation with time (R2 up to 0.94) were the crown-rump and occipito-snout lengths and the thoracic and biparietal diameters. No significant differences were observed in pregnancies with one or two fetuses in the period studied.     

Brucella-induced abortions and infection in bottlenose dolphins (Tursiops truncatus).

Two bottlenose dolphins (Tursiops truncatus) aborted fetuses that died as a result of Brucella infection. Brucella placentitis occurred in both cases. Infected placenta and vaginal/uterine fluids may transmit Brucella species to other cetaceans. In a third case, an identical organism was cultured from lung necropsy tissue of an adult female T. truncatus. Microbiology, specific polymerase chain reaction, and pulsed-field gel electrophoresis results supported the designation of an additional genomic group(s), Brucella delphini, for isolates adapted to T. truncatus. Current serologic diagnostic tests reliable for known Brucella species are unreliable in detecting dolphin brucellosis. Our findings, together with previous reports, suggest that dolphin brucellosis is a naturally occurring disease that can adversely impact reproduction in cetaceans. The zoonotic significance of cetacean brucellosis is unknown, although the disease has not been reported in people who have frequent contact with dolphins. Further studies on the zoonotic aspects, distribution, prevalence, virulence, and impact of this disease in cetaceans and other marine mammal species are needed.     

Sublingual squamous cell carcinoma in an Atlantic bottlenose dolphin (Tursiops truncatus).

A 22-yr-old captive-born Atlantic bottlenose dolphin (Tursiops truncatus) presented with a nonhealing sublingual mucosal ulcer that was diagnosed histologically as a squamous cell carcinoma, the first such report in a dolphin. The lesion was excised completely and has not recurred.     

Congenital diffuse hyperplastic goiter associated with perinatal mortality in 11 captive-born bottlenose dolphins (Tursiops truncatus).

Diffuse hyperplastic goiter was diagnosed by histopathology in 11 perinatal bottlenose dolphins (Tursiops truncatus) that died at four separate zoos and aquaria. Thyroid morphology of these animals was compared with the histologically normal thyroids of two stranded wild bottlenose dolphin calves, a neonate and a 2-mo-old calf. Histologic changes included reduced follicular luminal diameter, markedly reduced or absent luminal colloid, hypertrophy of follicular epithelium, and follicular dysplasia. The etiology of the thyroid gland lesion was not identified. Cause of death was not determined for most of these animals, but they were presumed to have died from metabolic derangements associated with the thyroid lesion, drowning, or dystocia.     

Dolphin pox: a skin disease of cetaceans.

Poxvirus has been identified morphologically from skin lesions in captive and free-ranging bottlenosed dolphins, Tursiops truncatus and a stranded Atlantic white-sided dolphin, Lagenorhynchus acutus. The lesions, commonly referred to as ring or pinhole lesions, appear as solitary or coalesced circular grey blemishes. Advanced ring lesions may take the form of black punctiform stippled patterns known as "tattoo". Histologically, the stratum externum is thickened, and there is ballooning degeneration and eosinophilic intractyoplasmic inclusions in the stratum intermedium. These includions contain virus particles which exhibit typical poxvirus morphology. Stress, environmental conditions and general health appear to play a major role in the clinical manifestation of dolphin pox.     

Comparison of the accuracy of two ultrasonographic measurements in predicting the parturition date in the bitch

OBJECTIVES: To compare the accuracy of two ultrasonographic measurements, inner chorionic cavity diameter and biparietal diameter, for the prediction of parturition date in small and medium size bitches. The effects of litter size and fetal sex ratio on the accuracy of the prediction were also investigated. METHODS: Ultrasonographic examinations were performed on pregnant bitches and, depending on the stage of pregnancy, the inner chorionic cavity diameter and biparietal diameter were measured. The mean value of the data of each structure was calculated and the predicted day of parturition was obtained by the application of equations derived from growth curves we previously developed for different size bitches. RESULTS: The parturition term was predicted with similar accuracy during early gestation, by the measurement of inner chorionic cavity diameter, as well as during late gestation, by the measurement of biparietal diameter. Litter size affected only biparietal diameter accuracy, whereas fetal sex ratio did not influence the estimation of the day of parturition. CLINICAL SIGNIFICANCE: Inner chorionic cavity diameter and biparietal diameter were equally reliable parameters in obtaining a reasonably accurate prediction of parturition date at any gestational period at which a bitch is examined.     

Infection with Brucella ceti and high levels of polychlorinated biphenyls in bottlenose dolphins (Tursiops truncatus) stranded in south-west England

Eight bottlenose dolphins (Tursiops truncatus) that stranded in Cornwall, south-west England, between June 2004 and December 2007 were examined using standardised postmortem examination and bacteriological methods. Evidence of Brucella species infection was found in four of these dolphins on culture. In addition, of the eight dolphins, four were positive and two were weakly positive for antibodies to Brucella species on serological analyses of pericardial and other fluids using a competitive ELISA and two indirect ELISAs. High or very high levels of the sum of 25 individual chlorobiphenyl congeners (∑25CBs) were also determined in blubber samples from two of the dolphins (45.5 and 446.6 mg/kg lipid weight).     

Paternity determination of captive bottlenose dolphins (Tursiops truncatus) using microsatellite DNA analysis

We applied previously published PCR primer pairs to amplify alleles at three polymorphic microsatellite loci to determine the genetic relationship of 6 bottlenose dolphins (Tursiops truncatus) that were living together in a Japanese aquarium. The three microsatellite loci were sufficient to determine the haplotype relationships of the six dolphins, which represented three different generations. It was confirmed that this genotyping method is simple and economical for assessing, establishing and maintaining genetic diversity in captive populations and will become a very effective technique for ex situ conservation in aquariums and zoos.     

Bronchoscopic and serologic diagnosis of Aspergillus fumigatus pulmonary infection in a bottlenose dolphin (Tursiops truncatus).

A 4-yr-old male bottlenose dolphin (Tursiops truncatus) developed an Aspergillus fumigatus pneumonia. Fungal elements were identified by cytology and microbiology from endoscopic bronchoalveolar lavage and brushings of a raised yellow endobronchial lesion. The results of qualitative immunodiffusion serology, a technique that identifies specific circulating antibodies to Aspergillus fumigatus, were suggestive of an active infection. The dolphin was treated with itraconazole for over 2 yr, which resulted in remission of clinical signs. Pneumonia caused by Aspergillus sp. accounts for the large majority of pulmonary mycoses in marine mammals. Bronchoscopy facilitated an early definitive diagnosis, accurate treatment, and remission.     

Disseminated histoplasmosis in an Atlantic bottlenose dolphin (Tursiops truncatus).

An approximately 37-yr-old female Atlantic bottlenose dolphin (Tursiops truncatus) died after a 4-mo illness characterized by intermittent anorexia, lethargy, mild neutrophilic leukocytosis, and mild nonregenerative anemia. At necropsy, the lungs were diffusely consolidated, and histopathology of the lungs revealed severe pneumonia with macrophages containing clusters of numerous yeast cells. Inflammatory lesions and yeast also were found in pulmonary, mediastinal, prescapular, and duodenal lymph nodes, spleen, liver, kidneys, urinary bladder, pancreas, right adrenal gland, and the pyloric stomach. Histomorphology, fungal culture, and polymerase chain reaction analysis indicated that the fungus was Histoplasma capsulatum var. capsulatum. This is the first report of histoplasmosis in a cetacean.     

ANATOMIC AND COMPUTED TOMOGRAPHIC ATLAS OF THE HEAD OF THE NEWBORN BOTTLENOSE DOLPHIN (TURSIOPS TRUNCATUS)

The head of a newborn dolphin (Tursiops truncatus), that died shortly after birth was imaged using computed tomography (CT). Gross cross-sectional slices of the head were compared with the CT images to identify normal structures of the cranium, brain, and respiratory and digestive pathways. Labelled transverse CT images of the dolphin head are presented sequentially as a reference for normal anatomy.     

Fatal necrotizing fasciitis and myositis in a captive common bottlenose dolphin (Tursiops truncatus) associated with Streptococcus agalactiae.

A common bottlenose dolphin (Tursiops truncatus) was presented for necropsy after acute onset of gastrointestinal signs and cutaneous lesions that rapidly progressed to death. Gross and microscopic findings were characterized by locally extensive severe necrohemorrhagic fasciitis and cellulitis, and severe necrotizing myositis in the head and dorsocranial thorax, with numerous disseminated gram-positive cocci. Streptococcus agalactiae was isolated from the lesions and from visceral organs (liver and lung), and it was identified by standard microbiology techniques. This communication is the first report of necrotizing fasciitis in a marine mammal associated with S. agalactiae.     

Hematologic, biochemical, and endocrine effects of dexamethasone on bottlenose dolphins (Tursiops truncatus).

Two Atlantic bottlenose dolphins (Tursiops truncatus) were given 0.11 mg/kg dexamethasone p.o., and complete blood count and serum chemistry analyses, including insulin, thyroxine (T4) adrenocorticotrophic hormone (ACTH), and cortisol level determinations, were performed at 0 hr, 24 hr, 36 hr, 48 hr, 7 days, and 17 days. Significant changes included neutrophilia, eosinopenia, lymphopenia, elevated insulin, and depressed ACTH and cortisol levels within 24 hr of dexamethasone administration. These effects were rapid, and values returned to normal within 48 hr.     

Scanning Electron Microscopy of the Orbital Harderian Gland in the Male Atlantic Bottlenose Dolphin (Tursiops truncatus)

The ultrastructure of the Harderian gland of Atlantic bottlenose dolphin ( Tursiops truncatus ) was examined by scanning electron microscopy (SEM). We found the following surface features: the typical round appearance of the ascinar glandular unit with a finely granular surface, a thin cortex and immediately below two types of cells: type I cells (characterized by small lipid vacuoles) and type II cells (characterized by large lipid vacuoles). It has been suggested that different cells forms represent a single cell type in varying activity states. Additionally, a coalescent tubular complex, a small balloon-like structures and large globular structures were observed. These structures may be reservoirs of secretion products.     

Real-time ultrasonography for determining the gestational age of ewes.

Seven Swedish peltsheep ewes were examined weekly by transabdominal ultrasound scanning from one-and-a-half to two months of gestation until parturition. By means of a simple regression analysis, the heart rate, the biparietal diameter of the skull and the diameter of the body trunk were correlated with fetal age, and it was concluded that these measurements could be used to estimate the age of the fetus.     

Cloning and sequencing of a bottle-nosed dolphin (Tursiops truncatus) interleukin-4-encoding cDNA.

Using polymerase chain reaction (PCR), a bottle-nosed dolphin (Tursiops truncatus) interleukin-4 (IL4) cDNA was cloned and sequenced. IL4 specific primers were based on the 5' and 3' untranslated regions of the human and murine IL4 gene. The dolphin IL4 cDNA is 528 base pairs in length and contains an open reading frame of 402 nucleotides coding an IL4 precursor of 133 amino acids, with the putative signal peptide of 24 amino acids. Analysis of the mature amino acid sequence shows three potential N-linked glycosylation sites and three disulfide bonds. Comparison of the predicted amino acid sequence shows that dolphin IL4 shares 77, 74, 58 and 41% identity with the bovine, ovine, human and mouse IL4s, respectively.