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.

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.

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.

Evaluation of ballistics euthanasia applied to stranded cetaceans using ethological and post-mortem computed tomography assessment.

Debilitated stranded cetaceans with low survival likelihood, may require euthanasia to avoid further suffering. Euthanasia can involve chemical or physical methods, including ballistics. Ballistics should cause instantaneous, permanent insensibility through brainstem disruption. Despite wide application, there is limited understanding of ballistics-related welfare outcomes. We opportunistically examined behaviour of three maternally-dependent cetaceans following shooting and the related cranial disruption post-mortem using computed tomography (PMCT). Our aim was to understand whether a 'humane death', i.e., euthanasia, was achieved. Each animal was shot using different projectile types: soft non-bonded, solid, and soft bonded. In two animals, insensibility was not immediately assessed following shooting, although both were reported as 'instantaneously insensible'. From our analysis, all animals displayed musculoskeletal responses to shooting, including peduncle stiffening and slack lower jaw, followed by musculature relaxation 24-, 10.3- and 20.8-seconds post-ballistics, respectively. The animal shot with a soft non-bonded projectile also displayed agonal convulsions and tail-lifting for 16-seconds post-shot; these were not observed for solid or soft bonded projectiles. PMCT findings indicated projectile disruption to the brainstem and/or spinal cord likely to cause near-instantaneous insensibility. However, extra-cranial wounding was also evident for the soft non-bonded projectile, highlighting potential for additional welfare compromise. Our results demonstrate that ballistics can achieve a relatively rapid death in young, stranded cetaceans, but careful equipment selection is required. To ensure a humane death, verification of insensibility must be undertaken immediately following shooting. Further studies should be undertaken to improve knowledge of appropriate procedures and equipment for euthanasia, ensuring humane deaths for compromised cetaceans.

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.

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.

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.

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.

Evaluation of the accuracy and reliability of two 3-dimensional sonography methods in volume measurement of small structures: an in vitro phantom study.

PURPOSE: To evaluate the accuracy and reliability of two 3-dimensional (3D) ultrasound imaging and measurement techniques in volume measurements using custom-made phantoms. METHODS: A total of 20 phantoms with irregular-shaped test objects of known volume (2-9.7 ml) were constructed. The phantoms were scanned using an automated mechanical 3D sonography technique and a free-hand 3D sonography technique. The volumes of the test objects were measured with "parallel planes" and "rotating planes" techniques, respectively. The measured volumes were compared with the actual volumes of the test objects. To evaluate inter- and intraoperator measurement variability, the phantoms were scanned twice by 2 different operators. RESULTS: Both the automated mechanical and the free-hand 3D sonography techniques were accurate and reliable. Automated mechanical 3D sonography with the parallel planes technique (accuracy, 81.5-83.4%; reproducibility, 91.1%; repeatability, 98.8-99.1%) was slightly more accurate and reliable than the free-hand rotating planes technique (accuracy, 74.7-84.2%; reproducibility, 88.4%; repeatability, 97.3-98%), but the differences were not statistically significant. CONCLUSION: Both the automated and freehand volume measurement techniques evaluated in this study are accurate and reliable.

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.

Three-dimensional ultrasound volumetric measurements: is the largest number of image planes necessary for outlining the region-of-interest?

Volumetric measurement is useful in the clinical practice as it can accurately assess the size of organs and lesions, which aids diagnosis and monitoring of treatment response. This in vitro study was undertaken to evaluate and compare the accuracy of three-dimensional ultrasound (3D-US) volumetric measurements using different numbers of image planes for outlining the region-of-interest. A total of 20 tissue phantoms (10 with regular and 10 with irregular tissue specimens) with known volume (2.2 - 14.7 ml) were constructed. The tissue phantoms were scanned with a commercially-available ultrasound machine in conjunction with a 3D add-on system and the volume of the tissue specimen of the phantoms was measured by manual outlining of the boundaries using different numbers of image plane (16, eight, four and two image planes). Results showed that there was a high accuracy in 3D-US volumetric measurements, when 16 and eight image planes were used (best-fit slope between measured and true volume: 0.9224-0.9761 and 0.9099-0.956, respectively). Results also showed that there was no significant difference in the volume measured with 16 and eight image planes for both phantoms with regular- and irregular-shaped tissue specimens (p > 0.05). The measurement differences between 16 and eight image planes ranged from -0.07 to 0.14 ml for phantoms with regular-shaped tissue and from -0.24 to 0.1 ml for phantoms with irregular-shaped tissue. 3D-US has a high accuracy in volumetric measurements of regular- and irregular-shaped structures. Although highest accuracy in volume measurements was found when 16 image planes were used, similar level of accuracy can be achieved with eight image planes but the measurement time can be reduced by 50%. Therefore, in 3D-US volumetric measurements, the largest number of image planes may not be necessary for outlining the region-of-interest.

A comparison of portable ultrasound and fully-equipped clinical ultrasound unit in the thyroid size measurement of the Indo-Pacific bottlenose dolphin.

Measurement of thyroid size and volume is a useful clinical parameter in both human and veterinary medicine, particularly for diagnosing thyroid diseases and guiding corrective therapy. Procuring a fully-equipped clinical ultrasound unit (FCUS) may be difficult in most veterinary settings. The present study evaluated the inter-equipment variability in dolphin thyroid ultrasound measurements between a portable ultrasound unit (PUS) and a FCUS; for both units, repeatability was also assessed. Thyroid ultrasound examinations were performed on 15 apparently healthy bottlenose dolphins with both PUS and FCUS under identical scanning conditions. There was a high level of agreement between the two ultrasound units in dolphin thyroid measurements (ICC = 0.859-0.976). A high intra-operator repeatability in thyroid measurements was found (PUS: ICC = 0.854-0.984, FCUS: ICC = 0.709-0.954). As a conclusion, no substantial inter-equipment variability was found between PUS and FCUS in dolphin thyroid size measurements under identical scanning conditions, supporting further application of PUS for quantitative analyses of dolphin thyroid gland in both research and clinical practices at aquarium settings.

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.