Malignant seminoma with metastasis, Sertoli cell tumor, and pheochromocytoma in a spotted dolphin (Stenella frontalis) and malignant seminoma with metastasis in a bottlenose dolphin (Tursiops truncatus)

Seminoma with metastasis was diagnosed in a spotted dolphin (Stenella frontalis) and an Atlantic bottlenose dolphin (Tursiops truncatus). Sertoli cell tumor and pheochromocytoma were also diagnosed in the spotted dolphin. The spotted and bottlenose dolphins were adult males that stranded and died on the coasts of northwest Florida and southeast North Carolina, respectively. Neoplasia is infrequently reported in cetaceans. This is the first report of seminoma, Sertoli cell tumor, and pheochromocytoma in a dolphin, the first report of three distinct neoplasms in a dolphin, and one of the few reports of malignant neoplasia in dolphins.     

Bartonella species detection in captive, stranded and free-ranging cetaceans

We present prevalence of Bartonella spp. for multiple cohorts of wild and captive cetaceans. One hundred and six cetaceans including 86 bottlenose dolphins (71 free-ranging, 14 captive in a facility with a dolphin experiencing debility of unknown origin, 1 stranded), 11 striped dolphins, 4 harbor porpoises, 3 Risso's dolphins, 1 dwarf sperm whale and 1 pygmy sperm whale (all stranded) were sampled. Whole blood (n = 95 live animals) and tissues (n = 15 freshly dead animals) were screened by PCR (n = 106 animals), PCR of enrichment cultures (n = 50 animals), and subcultures (n = 50 animals). Bartonella spp. were detected from 17 cetaceans, including 12 by direct extraction PCR of blood or tissues, 6 by PCR of enrichment cultures, and 4 by subculture isolation. Bartonella spp. were more commonly detected from the captive (6/14, 43%) than from free-ranging (2/71, 2.8%) bottlenose dolphins, and were commonly detected from the stranded animals (9/21, 43%; 3/11 striped dolphins, 3/4 harbor porpoises, 2/3 Risso's dolphins, 1/1 pygmy sperm whale, 0/1 dwarf sperm whale, 0/1 bottlenose dolphin). Sequencing identified a Bartonella spp. most similar to B. henselae San Antonio 2 in eight cases (4 bottlenose dolphins, 2 striped dolphins, 2 harbor porpoises), B. henselae Houston 1 in three cases (2 Risso's dolphins, 1 harbor porpoise), and untyped in six cases (4 bottlenose dolphins, 1 striped dolphin, 1 pygmy sperm whale). Although disease causation has not been established, Bartonella species were detected more commonly from cetaceans that were overtly debilitated or were cohabiting in captivity with a debilitated animal than from free-ranging animals. The detection of Bartonella spp. from cetaceans may be of pathophysiological concern.     

Evaluation of two-dimensional and three-dimensional ultrasound in the assessment of thyroid volume of the Indo-Pacific bottlenose dolphin (Tursiops aduncus)

The assessment of thyroid volume plays an indispensable role in the diagnosis and management of different thyroid diseases. The present study evaluates the accuracy of dolphin thyroid volume measurement as determined by four two-dimensional (2D) ultrasound methods (A-D), with a standard of reference using three-dimensional (3D) ultrasound. The measurement accuracy for different recognized thyroid configuration is also evaluated. Inter- and intraoperator variability of the measurement methods was determined. Thyroid ultrasound examinations were conducted in 16 apparently healthy Indo-Pacific bottlenose dolphins (Tursiops aduncus) with 2D and 3D ultrasound under identical scanning conditions. All 2D ultrasound measurement methods yielded high accuracies (79.9-81.3%) when compared with the 3D ultrasound measurement, and had high measurement reproducibility (77.6-86.2%) and repeatability (78.1-99.7%). For 2D ultrasound measurements, Methods A and B were more accurate and reliable than Methods C and D, regardless of thyroid configuration. Ultrasound is useful in the measurement of thyroid volume in bottlenose dolphins. For the first time, a reliable ultrasound scanning protocol for measuring dolphin thyroid volume was developed, which provides a means to establish a normative reference for the diagnosis of thyroid pathologies and to monitor the thyroid volume during the course of treatment in living dolphins. Key words: 3D ultrasound, Indo-Pacific bottlenose dolphin, thyroid volume measurement, Tursiops aduncus.     

Histological Structure of the Adrenal Gland of the Bottlenose Dolphin (Tursiops truncatus) and the Striped Dolphin (Stenella coeruleoalba) from the Adriatic Sea

The structure of the adrenal gland was studied in 11 bottlenose dolphins ( Tursiops truncatus ), and five striped dolphins ( Stenella coeruleoalba ). These species are legally protected in Croatia. All examined animals died of natural causes and were found stranded along eastern Adriatic coast. In both species the adrenal gland consists of a cortex and a medulla; the cortex is divided into three zones. Whereas in the bottlenose dolphin, there is a zona arcuata which contains columnar cells arranged in the form of arches; in the striped dolphin this zone is replaced by zona glomerulosa containing rounded clusters of polygonal cells. In both species, the zona fasciculata consists of radially oriented cords of polygonal cells, whereas in zona reticularis cells are arranged in branching and anastomosing cords. The adrenal medulla in both species contains dark, epinephrine-secreting cells and light norepinephrine-secreting cells. Epinephrine-secreting cells are localized in the outer part of the medulla, whereas norepinephrine-secreting cells are found in the inner part, arranged in clusters and surrounded by septa of thin connective tissue. The gland is surrounded by a thick connective-tissue capsule, from where thick trabeculae extend towards the interior. In the bottlenose dolphin, group of cells resembling both medullar and cortical cells can be seen within the capsule; whereas only groups of cells resembling cortical cells are found within the capsule of the striped dolphin. In the bottlenose dolphin invagination of the adrenal cortex into the medulla is obvious as well as medullary protrusions extending through cortex to the connective tissue capsule.     

The intestinal microbiome of an Indo-Pacific humpback dolphin (Sousa chinensis) stranded near the Pearl River Estuary,China

The mammalian intestinal microbiome is critical for host health and disease resistance. However, the cetacean intestinal microbiota remains relatively unexplored. By using high-throughput 16S rRNA gene sequencing, we analyzed intestinal bacterial samples from an Indo-pacific humpback dolphin (Sousa chinensis) stranded near the Pearl River Estuary in China. The samples included 3 anatomical regions (foregut, midgut, and rectum) and 2 anatomical locations (content and mucus). Our analyses revealed that the dolphin intestinal bacteria contained 139 operational taxonomic units (OTUs), dominated at the phyla level by Firmicutes (47.05% in the content; 94.77% in the mucus), followed by Bacteroidetes (23.63% in the content; 1.58% in the mucus) and Gammaproteobacteria (14.82% in the content; 2.05% in the mucus). The intestinal bacteria had a small core community (15 OTUs, accounting for 99.74% of the reads), some of which could be potentially pathogenic to both human and dolphins. As an alternative to sampling the dolphin intestinal bacteria, fecal sampling could be used. Additionally, function potentials such as, xenobiotics biodegradation, beta-lactam resistance, and human disease-related pathways, were detected in the dolphin intestinal bacteria. These findings provide the first baseline knowledge of the intestinal microbiome of the Indo-Pacific humpback dolphin, which may offer new insights into cetacean conservation by using microbial surveillance.     

Proliferative dermatitis associated with a novel alphaherpesvirus in an Atlantic bottlenose dolphin (Tursiops truncatus).

Herpesviruses and herpes-like viruses have been reported in only a small number of species of cetaceans, and, to date, clinical manifestations have been either as a life-threatening, disseminated infection or as a non-life-threatening dermatitis. A stranded juvenile Atlantic bottlenose dolphin, Tursiops truncatus, was admitted to the Dolphin and Whale Hospital for rehabilitation. On initial physical examination, the rostral skin had multifocal regions of hyperplasia, and the skin of the dorsum contained a large number of small papules. Histologically, epithelial hyperplasia was evident, and clusters of epithelial cells contained 5-15-microm intranuclear inclusion bodies. Transmission electron microscopic investigation revealed numerous 170-190-nm enveloped virions in both the intracellular spaces and the cytoplasm of epithelial cells, with numerous nucleocapsids noted in epithelial cell nuclei. Consensus primer polymerase chain reaction identified the presence of a novel herpesvirus associated with the lesions. Phylogenetic analysis of the deduced amino acid sequences of the herpesvirus DNA polymerase gene fragment showed it to align with alphaherpesvirus sequences from humans and domestic animals. Although clearly distinct, it was most closely related to two previously described alphaherpesviruses of dolphins. This case represents the first documentation of herpesvirus dermatitis in the Atlantic bottlenose dolphin.     

Calicivirus-induced vesicular disease in cetaceans and probable interspecies transmission

A calicivirus isolated from cetaceans is a new serotype designated cetacean calicivirus Tursiops-1 (CCV-Tur-1). It appears to have spread from an initially infected Atlantic bottlenose dolphin to a California sea lion, and was then carried by the sea lion to a second facility several miles away, where a second dolphin became infected and developed vesicular skin lesions that eroded, leaving shallow ulcers. Cetaceans and pinnipeds belong to separate orders, so this finding of interspecies transmission demonstrates the potentially broad host spectrum for yet another calicivirus.     

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.     

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.     

Prevalence of cutaneous pathological conditions and factors associated with the presence of skin wounds in working equids in tropical regions of Veracruz,Mexico

A wide spectrum of welfare issues is encountered on working equids. The objectives of this study were: (i) to determine the prevalence of cutaneous lesions in working equids in tropical regions of the North-Central of Veracruz State, Mexico, (ii) to identify the most prevalent cutaneous pathological conditions in this population of working equids, and (iii) to identify risk factors associated to the presence of work-related skin wounds. A number of 467 working equids presented for treatment at Donkey Sanctuary Mobile Clinic Program - National Autonomous University of Mexico from seven different villages in the North-Central region of Veracruz State were evaluated. Animals with signs of cutaneous pathology underwent a physical examination and samples were obtained of exudates, hair, and tissue for cytology examination, fungal cultures, and dermo-histopathology examination. The overall prevalence of cutaneous pathological conditions in working equids was 20.6 %. The prevalence per species was 22.6 % for horses, 18.2 % for donkeys, and 14.3 % for mules. The most common cause of skin lesions was trauma (abrasions, lacerations, and abscesses) followed in frequency by parasitic dermatitis and sarcoids. No strong associations among species were observed (P?>?0.20). Multivariate analysis showed that there was greater association between BCS and age with the presence of skin wounds (P?<?0.05). Equids who's BCS was rated as “moderate” as well as adult animals showed significantly more skin wounds compared with the young and the elderly (P?<?0.05). These findings can help to develop a better management practices to improve the welfare of equids in tropical regions of Veracruz, Mexico.     

Parasites from Indo-Pacific hump-backed dolphins (Sousa chinensis) and finless porpoises (Neophocaena phocaenoides) stranded in Hong Kong

Between 1993 and 1998, 28 Indo-Pacific hump-backed dolphins (Sousa chinensis) and 32 finless porpoises (Neophocaena phocoenoides) stranded in Hong Kong territorial waters were examined postmortem for parasites. The nematode Halocercus pingi was discovered in the lungs of one hump-backed dolphin and in 10 finless porpoises, typically within abscesses or granulomata, and they were frequently accompanied by a catarrhal exudate and lesions characteristic of pneumonia. Seven of the 10 finless porpoises were calves with substantial lungworm infections, and three were neonates with visible fetal folds and umbilical remnants, suggesting that H pingi is transferred to the neonate before birth or during lactation. Electron micrographs of H pingi should allow the nematode to be identified by other researchers. An ectoparasitic stalked barnacle (Xenobalanus globicipitis) was recovered from a finless porpoise, the first time that this species of barnacle has been recorded in Hong Kong's territorial waters.     

Temporal variation of the underwater soundscape in Jiaotou Bay,an Indo-Pacific humpback dolphin (Sousa chinensis) habitat off Hainan Island,China

The underwater soundscape is an important ecological element affecting numerous aquatic animals, in particular dolphins, which must identify salient cues from ambient ocean noise. In this study, temporal variations in the soundscape of Jiaotou Bay were monitored from February 2016 to January 2017, where a population of Indo-Pacific humpback dolphins (Sousa chinensis) has recently been a regular sighting. An autonomous acoustic recorder was deployed in shallow waters, and 1/3-octave band sound pressure levels (SPLs) were calculated with central frequencies ranging from 25 Hz to 40 kHz, then were grouped into 3 subdivided bands via cluster analysis. SPLs at each major band showed significant differences on a diel, fishing-related period, seasonal, and tidal phase scale. Anthropogenic noise generated by passing ships and underwater explosions were recorded in the study area. The fish and dolphin acoustic activities both exhibited diel and seasonal variations, but no tidal cycle patterns. A negative significant relationship between anthropogenic sound detection rates and dolphin detection rates were observed, and fish detection rates showed no effect on dolphin detection rates, indicating anthropogenic activity avoidance and no forced foraging in dolphins in the study area. The results provide fundamental insight into the acoustic dynamics of an important Indo-Pacific humpback dolphin habitat within a coastal area affected by a rapid increase in human activity, and demonstrate the need to protect animal habitat from anthropogenic noises.     

Behavior and salivary cortisol of captive dolphins (Tursiops truncatus) kept in open and closed facilities

Few studies have been carried out on the welfare of captive bottlenose dolphins (Tursiops truncatus). Of these, most include information on animals kept in closed facilities or pools. The aim of this study was to assess the welfare of captive bottlenose dolphins in open and closed facilities by measuring states of individual behavior and salivary cortisol concentrations. A total of 23 bottlenose dolphins were studied in 4 different dolphinaria. Dolphinaria A and B have closed facilities, whereas dolphinaria C and D have open facilities. A total of 152 hours of behavioral observations were analyzed using a combination of behavior and scan sampling to obtain information on individual time budgets. Salivary cortisol concentrations were measured using radioimmunoassay in 96 and 180 saliva samples of dolphins kept in closed and open facilities, respectively. In general, the results found that dolphins kept in open facilities spent less time floating (P < 0.05) and swimming in circular patterns than linear ones (P < 0.05) compared with dolphins in closed facilities. Dolphins kept in open facilities also had lower salivary cortisol concentrations than dolphins kept in closed facilities (P < 0.05). For this reason, we suggest that further research should include other welfare indicators such as reproductive function and health measurements to know more about the relationships between the design of pools and dolphin welfare.     

Development and validation of monoclonal and polyclonal antibodies for the detection of immunoglobulin G of bottlenose dolphins (Tursiops truncatus).

Antibodies directed against species-specific immunoglobulin G (IgG) have a broad range of applications in serologic and immunologic research and in the development of clinical assays. Validated anti-IgG antibodies for marine mammal species are in short supply. The objective of this study was to produce and validate antibodies with specificity for IgG of the common bottlenose dolphin (Tursiops truncatus). Bottlenose dolphin IgG was purified using protein G. Two mouse monoclonal antibodies and a rabbit polyclonal antibody were developed from mice and rabbits immunized with bottlenose dolphin IgG. The specificity of the monoclonal antibodies and the polyclonal antibody for bottlenose dolphin IgG was first verified by Western blot analysis and enzyme-linked immunosorbent assay (ELISA). For further validation, both monoclonal antibodies and the polyclonal antibody were incorporated in an indirect ELISA for the detection of the immune response of bottlenose dolphins to a vaccine antigen. Three bottlenose dolphins were immunized with a commercial Erysipelothrix rhusiopathiae vaccine, and serial blood samples were collected from all dolphins for measurement of levels of circulating antibodies. Seroconversion was observed in all 3 dolphins by use of both monoclonal antibodies and the polyclonal antibody. Circulating antibodies were detectable as early as 6 days after immunization in 1 dolphin. Peak antibody levels were detected 14 days after the immunization. The ability to detect seroconversion in all 3 immunized bottlenose dolphins firmly establishes the specificity of the monoclonal antibodies and the polyclonal antibody for IgG of the common bottlenose dolphin.     

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.     

Size and Shape Variability in the Skull of the Bottlenose Dolphin,Tursiops truncatus (Montagu, 1821)

The bottlenose dolphin (Tursiops truncatus) is a widely spread cetacean species. We present a quantitative analysis of age dimorphism variation in the skull of T. truncatus assessed by geometric morphometrics (GM) methods. Differences in size and shape of skulls were investigated using eight landmarks plotted on 2‐D images of ventral views of 14 museum specimens. The results of GM revealed differences in size, but not in shape. The left side appears more variable, which is probably an effect of telescoping.     

Sexual maturity,seasonal estrus,and gestation in female Indo-Pacific bottlenose dolphins Tursiops aduncus inferred from serum reproductive hormones

Reproductive hormones in serum concentrations of progesterone, estradiol, and testosterone in female Indo-Pacific bottlenose dolphins (Tursiops aduncus, n = 12) housed in Ocean Park Hong Kong were investigated in the present study. Results showed that, onset of puberty of captive Indo-Pacific bottlenose dolphins was at 5 years while sexual maturity was at 6. Average serum progesterone concentrations in non-pregnant sexually mature individuals was 0.33 (0.25–0.97) ng/mL (interquartile), significantly higher than in immature ones 0.26 (0.25–0.38) ng/mL. This study found significant difference in serum estradiol concentrations between individuals at the onset of puberty (9.5 ± 1.7 pg/mL, ±SD) and not (below detection limit 9 pg/mL). A slightly seasonal breeding pattern, with progesterone values tend to be higher from February to October (0.38 [0.25–1.07] ng/mL) was inferred. During pregnancy, serum progesterone concentrations range from 10.54 ± 8.74 ng/mL (indexed month post-conception [IMPC] 0) to 25.49 ± 12.06 ng/mL (IMPC 2), and display a bimodal pattern with 2 peaks in early- (25.49 ± 12.06 ng/mL, IMPC 2) and late-pregnancy (21.71 ± 10.25 ng/mL, IMPC 12), respectively. Serum estradiol concentrations can seldom be detected in early-pregnancy and increase constantly in mid- (9.45 ± 1.83 pg/mL) and late-pregnancy (11.88 ± 3.81 pg/mL), with a spike (15.45 ± 6.78 pg/mL) 1 month prior to delivery. Serum testosterone concentrations elevate significantly in IMPC 7 (0.36 ± 0.10 ng/mL) compared to other months (0.16 ± 0.10 ng/mL) of the year. The present study provides normal concentration profiles for some reproductive hormones in female Indo-Pacific bottlenose dolphins and can contribute to the breeding monitoring of this species. Also, our study would shed further light on the reproductive physiology of small cetaceans.     

Disseminated panniculitis in a bottlenose dolphin (Tursiops truncatus) due to Mycobacterium chelonae infection

A 14-year-old female bottlenose dolphin was diagnosed with mycobacterial panniculitis based on punch biopsy specimens. The necropsy revealed numerous pyogranulomas in the blubber, as well as marked acute multifocal necrosuppurative pneumonia and lymphadenitis. In addition, the animal had marked scoliosis, which had first been noted clinically in the dolphin at about 1 mo of age. The animal had been treated with low-dose dexamethasone for approximately the last 19 mo to reduce perceived discomfort from spondyloarthritis and with the progestational agent altrogenest for approximately 8 yr to prevent pregnancy. Acid-fast positive bacilli were detected in the dermis but not in lung or lymph nodes. Mycobacterium chelonae was isolated from pooled skin, lung, and peripheral lymph-node tissue. Mycobacterial infection may be considered as a differential diagnosis in bottlenose dolphins with generalized cutaneous inflammation, particularly if chronic steroid and progesterone treatments were administered, both of which may have an immunomodulatory effect.     

Brucellosis associated with stillbirth in a bottlenose dolphin in Australia

A captive adult female bottlenose dolphin presented with stillbirth. The placenta appeared oedematous. No other gross lesions were evident in the placenta or the stillborn calf. Histopathology revealed mild multifocal placentitis and foetal encephalitis. Brucella sp. was isolated from lung, liver, spleen and kidney. Sequence and phylogenetic analysis demonstrated this organism to be most similar to Brucella ceti sequence type (ST) 27. Brucella sp. DNA was detected in formalin-fixed paraffin-embedded placenta and brain by real-time PCR using primers targeting the IS711 gene. Immunohistochemical staining revealed Brucella sp. antigen in placental inflammation. This is the first report of isolation of Brucella sp. from a marine mammal in the Southern Hemisphere and the first report of marine Brucella-associated disease in Australia.     

Toxoplasmosis in a Guiana dolphin (Sotalia guianensis) from Paraná, Brazil

This study describes toxoplasmosis in a by caught Guiana dolphin (Sotalia guinensis) from Paranaguá Bay, Paraná, Brazil. Interstitial pneumonia, multisystemic arteritis, multifocal adrenalitis and hepatitis were the primary lesions observed. These tissues had moderate to severe necrosis and mononuclear cells infiltration usually surrounded by tachyzoites and tissue cysts. Moderate lymphoid depletion was evident in the spleen. Toxoplasma gondii was positive by immunohistochemical and ultrastructural evaluation. Furthermore, the animal was negative for Morbillivirus by immunohistochemistry and had low levels of persistent organochlorines. There is evidence of environmental changes in the Paranaguá Bay that could justify the occurrence of toxoplasmosis in Guiana dolphin. The sewage run-off from main urban areas and the presence of domestic and wild felids in areas surrounding the bay could be a source of T. gondii oocysts from land to sea. Based on its habitat, the authors recommend this dolphin species as sentinels for the health of bays and estuaries where they occur.