Temporal Trends and Suspect Screening of Halogenated Flame Retardants and Their Metabolites in Blubbers of Cetaceans Stranded in Hong Kong Waters during 2013-2020.

Halogenated flame retardants (HFRs) are a large class of chemical additives intended to meet flammability safety requirements, and at present, they are ubiquitous in the environment. Herein, we conducted the target analysis and suspect screening of legacy and novel HFRs and their metabolites in the blubber of finless porpoises (Neophocaena phocaenoides; n = 70) and Indo-Pacific humpback dolphins (Sousa chinensis; n = 35) stranded in Hong Kong, a coastal city in the South China Sea, between 2013 and 2020. The average concentrations of total target HFRs (ΣHFRs) were 6.48 × 103 ± 1.01 × 104 and 1.40 × 104 ± 1.51 × 104 ng/g lipid weight in porpoises and dolphins, respectively. Significant decreasing temporal trends were observed in the concentrations of tetra-/penta-/hexa-bromodiphenyl ethers (tetra-/penta-/hexa-BDEs) in adult porpoises stranded from 2013-2015 to 2016-2020 (p < 0.05), probably because of their phasing out in China. No significant difference was found for the concentrations of decabromodiphenyl ether and hexabromocyclododecane, possibly due to their exemption from the ban in China until 2025 and 2021, respectively. Eight brominated compounds were additionally identified via suspect screening. A positive correlation was found between the concentrations of tetra-BDE and methyl-methoxy-tetra-BDE (Me-MeO-tetra-BDE) (p < 0.05), indicating that the metabolism of tetra-BDE may be a potential source of Me-MeO-tetra-BDE in marine mammals.

An Indo-Pacific humpback dolphin (Sousa chinensis) severely injured by vessel collision: live rescue at sea, clinical care, and postmortem examination using a virtopsy-integrated approach.

BACKGROUND: Vessel collision induces blunt and sharp force traumas to aquatic animals and is a leading anthropogenic impact affecting cetaceans worldwide. Vessel collision is an important threat affecting vulnerable coastal cetaceans such as the Indo-Pacific humpback dolphins (Sousa chinensis) which reside in coastal waters of Hong Kong amongst heavy marine traffic. CASE PRESENTATION: A severely injured subadult S. chinensis was sighted in the waters off southwestern Hong Kong with four gaping incision wounds on its dorsum. It was in poor body condition and seemed unable to use the fluke effectively. The deepest wound located at the caudal peduncle near the base of the fluke and exposed the underlying fractured caudal vertebrae. The dolphin was monitored in the field over three weeks and eventually captured for medical intervention as veterinary assessment indicated progressive and life-threatening deterioration. During rehabilitation, the dolphin demonstrated initial signs of improvement over the first 36 hours as supported by diagnostic tests but then deteriorated rapidly. It was humanely euthanised after three days of rehabilitation. Postmortem investigation was carried out using virtopsy (postmortem computed tomography and magnetic resonance imaging) and conventional necropsy, with special attention to the traumatic musculoskeletal injuries caused by vessel collision and also revealed acute gastrointestinal compromise and respiratory disease that further hampered the rehabilitation. CONCLUSION: In cetaceans, the prognosis for recovery from injuries caused by vessel collision depends on the extent, location, and gravity of the injuries (i.e., superficial, deep, penetrating, blunt vs. sharp, fresh vs. septic), as well as the health status of the individual and its ability to respond to the insult. Injuries extending deep into the vertebral column may lead to delayed death and associated welfare issues. The prognosis of this case was likely poor given the severity and location of the injuries, but the attempted rehabilitation and postmortem investigation provided valuable insights for clinical management if similar cases are encountered in the future. Being able to non-invasively assess and document traumatic injuries and other pathologies, diagnostic imaging is particularly useful in the clinical assessment and postmortem investigation (virtopsy) of cases with vessel-induced injuries.

First Unmanned Aerial Vehicle Observation of Epimeletic Behavior in Indo-Pacific Humpback Dolphins.

Epimeletic behavior has been reported in various species of cetaceans and sometimes in wild populations during vessel-based surveys. Epimeletic behavior in cetaceans involves complex social interactions which have been described using observational and acoustic studies. The recent advances in unmanned aerial vehicle (UAV) technology allowed its application in wildlife research and frequently in cetaceans in conjunction with vessel-based surveys. This article is the first report of intraspecific epimeletic behavior of Indo-Pacific humpback dolphins (Sousa chinensis) in Hong Kong waters using a combination of UAV- and vessel-based photography. Using both techniques, we were able to observe and qualitative analyze the individual body condition, group behavior, and swimming pattern during the epimeletic event. This study highlighted that UAVs can be used to observe the complex social behaviors and interactions of cetaceans from the aerial angle while keeping a minimal level of disturbance to the animals. Aerial footage can also be quantitatively analyzed to provide further insights on the group behaviors of cetaceans. The application allows efficient assessment of health, behavior, and ecology of wild animals, offering valuable opportunities for researchers working on free-ranging populations.

Characterisation of Crassicauda fuelleborni nematode infection in Indo-Pacific finless porpoises (Neophocaena phocaenoides) using postmortem computed tomography.

Nematodes of the genus Crassicauda are parasites that infect various body tissues of cetaceans, including the mammary glands which can influence the reproductive output and hence threaten the survival of endangered cetacean populations. In this study, postmortem computed tomography (PMCT) was used to characterise lesions related to Crassicauda fuelleborni infections in stranded Indo-Pacific finless porpoises (Neophocaena phocaenoides) from Hong Kong waters. Using PMCT and subsequently verified by conventional necropsy, Crassicauda-related lesions were found in 52% of finless porpoises examined (n = 13/25), including both males and females. These parasitic lesions were mostly located in the ventral abdominal muscles in both sexes and situated in proximity to the mammary glands in females. C. fuelleborni infections were also found in the male reproductive organs, which to our knowledge have not been reported in this cetacean species previously. PMCT characteristics of the lesions were also correlated with the gross appearance observed at necropsy and the chronicity of the parasitic infections. In conclusion, this study established the use of virtopsy, particularly PMCT, to characterise C. fuelleborni infections in stranded finless porpoises for the first time, which is non-invasive and can be used prior to conventional necropsy to aid disease diagnosis and targeted sampling. This technique can be extended to other species of cetaceans and parasites, as well as being used in the retrospective analysis of past PMCT scans to deepen our understanding of the prevalence, health impacts, and ecological implications of parasitic infection in cetaceans.

Visual Assessment of Contusion-Like Lesions Caused by Live Sharksucker (Echeneis naucrates) Attachment in an Indo-Pacific Humpback Dolphin (Sousa chinensis).

We observed contusion-like lesions caused by live sharksucker (Echeneis naucrates) attachment in an Indo-Pacific humpback dolphin (Sousa chinensis). Our report assists in differentiating epidermal lesions for visual health assessment in this vulnerable species and other cetaceans, and this adds new host and epibiont records for E. naucrates and S. chinensis, respectively.

Image Rendering Techniques in Postmortem Computed Tomography: Evaluation of Biological Health and Profile in Stranded Cetaceans.

With 6 years of experience in implementing virtopsy routinely into the Hong Kong cetacean stranding response program, standardized virtopsy procedures, postmortem computed tomography (PMCT) acquisition, postprocessing, and evaluation were successfully established. In this pioneer cetacean virtopsy stranding response program, PMCT was performed on 193 stranded cetaceans, providing postmortem findings to aid necropsy and shed light on the biological health and profile of the animals. This study aimed to assess 8 image rendering techniques in PMCT, including multiplanar reconstruction, curved planar reformation, maximum intensity projection, minimum intensity projection, direct volume rendering, segmentation, transfer function, and perspective volume rendering. Illustrated with practical examples, these techniques were able to identify most of the PM findings in stranded cetaceans and served as a tool to investigate their biological health and profile. This study could guide radiologists, clinicians and veterinarians through the often difficult and complicated realm of PMCT image rendering and reviewing.

Photogrammetric Three-dimensional Modeling and Printing of Cetacean Skeleton using an Omura's Whale Stranded in Hong Kong Waters as an Example.

The preparation of cetacean, in particular baleen whale, skeletons presents a great challenge due to their high lipid content and uncommon size. Documentation of the skeletal morphology is important to produce accurate and reliable models for both research and educational purposes. In this paper, we used a 10.8-meter long Omura's whale stranded in Hong Kong waters in 2014 as an example for the illustration. This rare and enormous specimen was defleshed, macerated, and sun-dried to yield the skeleton for research and public display. Morphology of each bone was then documented by photogrammetry. The complex contour of the skeleton made automated photoshoot inadequate and 3 manual methods were used on bones of different sizes and shapes. The captured photos were processed to generate three-dimensional (3D) models of 166 individual bones. The skeleton was printed half-size with polylactic acid for display purposes, which was easier to maintain than the actual cetacean bones with high residual fat content. The printed bones reflected most anatomical features of the specimen, including the bowing out rostral region and the caudal condylar facet that articulated with Ce1, yet the foramina on the parieto-squamosal suture, which are diagnostic character of Balaenoptera omurai, and an indented groove on the frontal bone at the posterior end of the lateral edge were not clearly presented. Extra photoshoots or 3D surface scanning should be performed on areas with meticulous details to improve precision of the models. The electronic files of the 3D skeleton were published online to reach a global audience and facilitate scientific collaboration among researchers worldwide.

Diagnosis of atlanto-occipital dissociation: Standardised measurements of normal craniocervical relationship in finless porpoises (genus Neophocaena) using postmortem computed tomography.

Due to the different craniocervical structures in humans and cetaceans, a standardised method assessing the normal craniocervical relationship in cetaceans is lacking, causing difficulties in defining the presence of atlanto-occipital dissociation (AOD) in cetaceans. The present study aimed to 1) describe a novel standardised method of determining the normal craniocervical relationships, and 2) define the 95% accuracy range of the normal craniocervical relationship in finless porpoises (genus Neophocaena), that allowed AOD diagnosis. Fifty-five out 83 stranded or by-caught finless porpoise carcasses were analyzed in term of their craniocervical relationship in dorsal-ventral and medial-lateral dimension, using postmortem computed tomography measurements. The normal craniocervical relationship in both dorsal-ventral (mean BD/OV: 0.87 ± 0.24 [2 SD]) and medial-lateral dimension (mean VR/VL: 0.98 ± 0.17 [2 SD]) was first defined. The 95% accuracy ranges of the normal craniocervical relationship in dorsal-ventral (0.63-1.11) and medial-lateral dimension (0.82-1.15) were proposed. The baseline ranges could facilitate AOD assessment, and provide an objective means of record for AOD related injury and death of cetaceans caused by anthropogenic trauma. The technique developed may be applied to live cetaceans with abnormal craniocervical relationship to aid diagnosis and guide corrective therapy.

Database documentation of marine mammal stranding and mortality: current status review and future prospects.

Databases are systematic tools to archive and manage information related to marine mammal stranding and mortality events. Stranding response networks, governmental authorities and non-governmental organizations have established regional or national stranding networks and have developed unique standard stranding response and necropsy protocols to document and track stranded marine mammal demographics, signalment and health data. The objectives of this study were to (1) describe and review the current status of marine mammal stranding and mortality databases worldwide, including the year established, types of database and their goals; and (2) summarize the geographic range included in the database, the number of cases recorded, accessibility, filter and display methods. Peer-reviewed literature was searched, focussing on published databases of live and dead marine mammal strandings and mortality and information released from stranding response organizations (i.e. online updates, journal articles and annual stranding reports). Databases that were not published in the primary literature or recognized by government agencies were excluded. Based on these criteria, 10 marine mammal stranding and mortality databases were identified, and strandings and necropsy data found in these databases were evaluated. We discuss the results, limitations and future prospects of database development. Future prospects include the development and application of virtopsy, a new necropsy investigation tool. A centralized web-accessed database of all available postmortem multimedia from stranded marine mammals may eventually support marine conservation and policy decisions, which will allow the use of marine animals as sentinels of ecosystem health, working towards a 'One Ocean-One Health' ideal.

Accuracy and reliability of cetacean cranial measurements using computed tomography three dimensional volume rendered images.

Computed tomography (CT) has become more readily available for post-mortem examination, offering an alternative to cetacean cranial measurements obtained manually. Measurement error may result in possible variation in cranial morphometric analysis. This study aimed to evaluate the accuracy and reliability of cetacean cranial measurements obtained by CT three-dimensional volume rendered images (3DVRI). CT scans of 9 stranded cetaceans were performed. The acquired images were reconstructed using bone reconstruction algorithms. The reconstructed crania obtained by 3DVRI were visualized after excluding other body structures. Accuracy of cranial measurements obtained by CT 3DVRI was evaluated by comparing with that obtained by manual approach as standard of reference. Reproducibility and repeatability of cranial measurements obtained by CT 3DVRI were evaluated using intraclass correlation coefficient (ICC). The results demonstrated that cranial measurements obtained by CT 3DVRI yielded high accuracy (88.05%- 99.64%). High reproducibility (ICC ranged from 0.897 to 1.000) and repeatability (ICC ranged from 0.919 to 1.000 for operator 1 and ICC range from 0.768 to 1.000 for operator 2) were observed in cranial measurements obtained by CT 3DVRI. Therefore, cranial measurements obtained by CT 3DVRI could be considered as virtual alternative to conventional manual approach. This may help the development of a normative reference for current cranial maturity and discriminant analysis studies in cetaceans.

Effect of Photoperiod Extension on the Testicular Sonographic Appearance and Sexual Behavior of Captive Yangtze Finless porpoise (Neophocaena asiaeorientalis asiaeorientalis).

Xueying Yu, Yujiang Hao, Brian CW Kot, and Ding Wang (2016) Photoperiod plays an important role in the seasonal regulation of reproduction in mammals. In the present study, we examined the effect of an extended photoperiod (light/dark: 14 h/10 h) during the usual sexually inactive phase (Jan-Mar) on testicular sonographic appearance and sexual behaviors of two captive Yangtze finless porpoise (Neophocaena asiaeorientalis asiaeorientalis, YFP). During the extended photoperiod treatment, the parenchyma pixel intensity, cross- sectional area of testicular sonographic images, as well as the frequency of sexual behaviors of the two male porpoises increased significantly compared to the control treatment. Parenchyma pixel intensity, the cross- sectional area of testicular sonographic images and the frequency of sexual behaviors of the two captive males all increased in response to extended photoperiod. These results may suggest that photoperiod is a potent factor regulating the reproductive seasonality in YFPs.

Sonographic appearance of thyroid glands in patients treated with intensity-modulated radiotherapy or conventional radiotherapy for nasopharyngeal carcinoma.

BACKGROUND: This study aimed to investigate the sonographic appearances of the thyroid glands in nasopharyngeal carcinoma (NPC) patients whose cervical lymph nodes were treated with conventional radiotherapy (RT) or intensity-modulated radiotherapy (IMRT). The post-RT sonographic appearances of the thyroid glands in NPC patients were also correlated with the thyroid function. METHODS: One hundred and three NPC patients who had completed RT of cervical lymph nodes using the anterior cervical field, 30 NPC patients who had completed RT of cervical lymph nodes using IMRT, and 61 healthy subjects were included in the study. Thyroid glands were sonographically assessed for their size, echogenicity, vascularity, and internal architecture. Thyroid function tests were also performed on each subject. RESULTS: In comparison with the patients with abnormal thyroid function, the thyroid glands of the patients with normal thyroid function tended to be homogeneous and to have greater volume and echogenicity index (p < 0.05). Compared with those of the healthy subjects, the thyroid glands of patients previously treated with IMRT and those treated with the anterior cervical field showed significantly lower thyroid volume, lower incidence and number of nodules, and higher vascularity index (p < 0.05). CONCLUSIONS: The patient's history of previous RT should be taken into consideration in the sonographic examination of the thyroid gland post-RT. © 2014 Wiley Periodicals, Inc. J Clin Ultrasound 43:210-223, 2015.

Innervation patterns of sea otter (Enhydra lutris) mystacial follicle-sinus complexes.

Sea otters (Enhydra lutris) are the most recent group of mammals to return to the sea, and may exemplify divergent somatosensory tactile systems among mammals. Therefore, we quantified the mystacial vibrissal array of sea otters and histologically processed follicle-sinus complexes (F - SCs) to test the hypotheses that the number of myelinated axons per F - SC is greater than that found for terrestrial mammalian vibrissae and that their organization and microstructure converge with those of pinniped vibrissae. A mean of 120.5 vibrissae were arranged rostrally on a broad, blunt muzzle in 7-8 rows and 9-13 columns. The F-SCs of sea otters are tripartite in their organization and similar in microstructure to pinnipeds rather than terrestrial species. Each F-SC was innervated by a mean 1339 ± 408.3 axons. Innervation to the entire mystacial vibrissal array was estimated at 161,313 axons. Our data support the hypothesis that the disproportionate expansion of the coronal gyrus in somatosensory cortex of sea otters is related to the high innervation investment of the mystacial vibrissal array, and that quantifying innervation investment is a good proxy for tactile sensitivity. We predict that the tactile performance of sea otter mystacial vibrissae is comparable to that of harbor seals, sea lions and walruses.

Feeding kinematics, suction, and hydraulic jetting performance of harbor seals (Phoca vitulina).

The feeding kinematics, suction and hydraulic jetting capabilities of captive harbor seals (Phoca vitulina) were characterized during controlled feeding trials. Feeding trials were conducted using a feeding apparatus that allowed a choice between biting and suction, but also presented food that could be ingested only by suction. Subambient pressure exerted during suction feeding behaviors was directly measured using pressure transducers. The mean feeding cycle duration for suction-feeding events was significantly shorter (0.15±0.09 s; P<0.01) than biting feeding events (0.18±0.08 s). Subjects feeding in-water used both a suction and a biting feeding mode. Suction was the favored feeding mode (84% of all feeding events) compared to biting, but biting comprised 16% of feeding events. In addition, seals occasionally alternated suction with hydraulic jetting, or used hydraulic jetting independently, to remove fish from the apparatus. Suction and biting feeding modes were kinematically distinct regardless of feeding location (in-water vs. on-land). Suction was characterized by a significantly smaller gape (1.3±0.23 cm; P<0.001) and gape angle (12.9±2.02°), pursing of the rostral lips to form a circular aperture, and pursing of the lateral lips to occlude lateral gape. Biting was characterized by a large gape (3.63±0.21 cm) and gape angle (28.8±1.80°; P<0.001) and lip curling to expose teeth. The maximum subambient pressure recorded was 48.8 kPa. In addition, harbor seals were able to jet water at food items using suprambient pressure, also known as hydraulic jetting. The maximum hydraulic jetting force recorded was 53.9 kPa. Suction and hydraulic jetting where employed 90.5% and 9.5%, respectively, during underwater feeding events. Harbor seals displayed a wide repertoire of behaviorally flexible feeding strategies to ingest fish from the feeding apparatus. Such flexibility of feeding strategies and biomechanics likely forms the basis of their opportunistic, generalized feeding ecology and concomitant breadth of diet.

Stereology of the thyroid gland in Indo-Pacific bottlenose dolphin (Tursiops aduncus) in comparison with human (Homo sapiens): quantitative and functional implications.

The mammalian thyroid gland maintains basal metabolism in tissues for optimal function. Determining thyroid volume is important in assessing growth and involution. Volume estimation is also important in stereological studies. Direct measurements of colloid volume and nuclear-to-cytoplasmic ratio of the follicular cells may provide important information about thyroid gland function such as hormone storage and secretion, which helps understand the changes at morphological and functional levels. The present study determined the colloid volume using simple stereological principle and the nuclear-to-cytoplasmic ratio of 4 Indo-Pacific bottlenose dolphins and 2 human thyroid glands. In both dolphin and human thyroid glands, the size of the follicles tended to be quite variable. The distribution of large and small follicles within the thyroid gland was also found to be random in both the dolphin and human thyroid gland; however, the size of follicles appeared to decrease as a function of increasing age in the dolphin thyroid gland. The mean colloid volume of the dolphin thyroid gland and human thyroid gland was 1.22×10(5) µm(3) and 7.02×10(5) µm(3) respectively. The dolphin and human subjects had a significant difference in the mean colloid volume. The mean N/C ratio of the dolphin thyroid follicular epithelia and human follicular epithelia was 0.50 and 0.64 respectively. The dolphin and human subjects had a significant difference in the mean N/C ratio. This information contributes to understanding dolphin thyroid physiology and its structural adaptations to meet the physical demands of the aquatic environment, and aids with ultrasonography and corrective therapy in live subjects.

Three-dimensional elastography for cervical lymph node volume measurements: a study to investigate feasibility, accuracy and reliability.

This study investigated the feasibility of using three-dimensional (3-D) elastography in measuring cervical lymph node volume and compared the accuracy and reliability of 3-D elastography and 3-D grayscale ultrasound in measurement of ill-defined cervical nodes. Eighteen porcine lymph nodes from the neck were embedded in tissue-mimicking phantoms and scanned with the two ultrasound techniques. Ultrasound measurements were compared with the volume determined by water-displacement method to evaluate measurement accuracy. Inter-observer reproducibility and intra-observer repeatability of measurements were evaluated. Four patients with enlarged neck nodes were included to evaluate intra-observer repeatability of ultrasound measurements. Results demonstrated that lymph nodes that appeared ill-defined on grayscale ultrasound showed well-defined boundaries on elastography. 3-D elastography has higher measurement accuracy (84.2%), reproducibility (intraclass correlation coefficient, ICC = 0.909) and repeatability (ICC = 0.964-0.988) than does 3-D grayscale ultrasound (62.2%, 0.777 and 0.863-0.906 respectively). As a conclusion, 3-D elastography is accurate and reliable in volume measurement of ill-defined lymph nodes and has potential for accurate assessment of lymph node volume.

Ultrasonographic assessment of the thyroid gland and adjacent anatomic structures in Indo-Pacific bottlenose dolphins (Tursiops aduncus).

OBJECTIVE: To evaluate the use of ultrasonography for thyroid gland assessment in healthy Indo-Pacific bottlenose dolphins (Tursiops aduncus), describe the ultrasonographic appearance of the thyroid gland and adjacent anatomic structures, and identify potential associations between variations in thyroid gland morphology and demographic features in this species. ANIMALS: 18 captive Indo-Pacific bottlenose dolphins. PROCEDURES: 1,404 ultrasonographic examinations of the thyroid gland and adjacent anatomic structures (eg, cervical lymph nodes, musculature, and vasculature) were performed during the > 3-year study period. Shape, echogenicity, and homogeneity of thyroid glands were assessed, and glands were categorized into morphological configurations on the basis of results of 2-D and 3-D ultrasonographic evaluation. Associations between demographic factors and thyroid gland morphology were assessed. RESULTS: Thyroid lobes appeared elliptical or fusiform in the transverse scan plane and round to oval in longitudinal scan planes; morphologically, glands comprised 2 lobes joined by an isthmus or a roughly diamond-shaped structure located on the ventral surface of the trachea. Major blood vessels and cervical lymph nodes were identified. Thyroid parenchyma was typically uniform and homogeneous, with echogenic reticulations and well-defined borders. Thyroid glands were hypoechoic or isoechoic relative to the sternocephalicus muscle; echogenicity was greater in adolescents than in adults. Thyroid gland volume differed between sexes, between sexually mature and immature dolphins, and among age groups and was positively correlated with body length and weight. CONCLUSIONS AND CLINICAL RELEVANCE: Ultrasonography provided a reliable and repeatable method for evaluation of thyroid glands and adjacent anatomic structures in live dolphins.

Elastic modulus of muscle and tendon with shear wave ultrasound elastography: variations with different technical settings.

Standardization on Shear wave ultrasound elastography (SWUE) technical settings will not only ensure that the results are accurate, but also detect any differences over time that may be attributed to true physiological changes. The present study evaluated the variations of elastic modulus of muscle and tendon using SWUE when different technical aspects were altered. The results of this study indicated that variations of elastic modulus of muscle and tendon were found when different transducer's pressure and region of interest (ROI)'s size were applied. No significant differences in elastic modulus of the rectus femoris muscle and patellar tendon were found with different acquisition times of the SWUE sonogram. The SWUE on the muscle and tendon should be performed with the lightest transducer's pressure, a shorter acquisition time for the SWUE sonogram, while measuring the mean elastic modulus regardless the ROI's size.

Sonographic evaluation of thyroid morphology during the normal estrous cycle in the Indo-Pacific bottlenose dolphin (Tursiops aduncus).

Physiologic changes occurring in the thyroid in response to the estrous cycle have been noted in companion animals. However, in bottlenose dolphins, the influence of different reproductive states on thyroid morphology remains unclear. Sonography was used to evaluate the variations of thyroid morphology for nine consecutive estrous cycles of four sexually mature, female Indo-Pacific bottlenose dolphins (Tursiops aduncus). The estrous cycle was divided into two phases for evaluation: the follicular and luteal phases. To compare changes associated with sex, thyroid volumes were measured in the different phases during the estrous cycle of female dolphins and were compared to the ultrasound examinations of the four male dolphins measured over a 2-mo period. In males, there was no significant difference in thyroid volume during the study period (P > 0.05). The thyroid volume measured in combining all estrous cycle phases of the female dolphins was significantly larger than that measured in the male dolphins (P < 0.05). A difference in thyroid volume during the estrous cycle was observed, with the thyroid volume during the follicular phase significantly smaller (P < 0.05) than that of the luteal phase, and is possibly related to the influence of female sex steroids. Thyroid volume variability during estrus should, therefore, be taken into account when examining the thyroid gland of female dolphins.

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.