Anatomical and volumetric description of the guiana dolphin (Sotalia guianensis) brain from an ultra-high-field magnetic resonance imaging.

The Guiana dolphin (Sotalia guianensis) is a common species along Central and South American coastal waters. Although much effort has been made to understand its behavioral ecology and evolution, very little is known about its brain. The use of ultra-high field MRI in anatomical descriptions of cetacean brains is a very promising approach that is still uncommon. In this study, we present for the first time a full anatomical description of the Guiana dolphin's brain based on high-resolution ultra-high-field magnetic resonance imaging, providing an exceptional level of brain anatomical details, and enriching our understanding of the species. Brain structures were labeled and volumetric measurements were delineated for many distinguishable structures, including the gray matter and white matter of the cerebral cortex, amygdala, hippocampus, superior and inferior colliculi, thalamus, corpus callosum, ventricles, brainstem and cerebellum. Additionally, we provide the surface anatomy of the Guiana dolphin brain, including the labeling of main sulci and gyri as well as the calculation of its gyrification index. These neuroanatomical data, absent from the literature to date, will help disentangle the history behind cetacean brain evolution and consequently, mammalian evolution, representing a significant new source for future comparative studies.

Apneustic anesthesia ventilation improves pulmonary function in anesthetized bottlenose dolphins (Tursiops truncatus).

Introduction: Use of mechanical ventilation during general anesthesia is a necessary practice in the anesthetization of small cetaceans as spontaneous ventilation fails to provide adequate gas exchange. Currently available methods of ventilation do not account for the intermittent breathing strategy of representative species within this infraorder of fully aquatic mammals and may have a significant effect on cardiac and respiratory physiology. Methods: To understand the impact of mechanical ventilation on cardiopulmonary function in one small species of cetacean, the bottlenose dolphin (Tursiops truncatus), we compared controlled mechanical ventilation (CMV) to a novel ventilation method known as apneustic anesthesia ventilation (AAV). AAV simulates the normal inspiratory breath-hold pattern of dolphins. Ten anesthetic procedures (dental procedure, n = 9; bronchoscopy, n = 2) were performed on nine dolphins (age range: 10-42 years; mean = 32 years; median = 37 years; female = 3, 40%; male = 6, 60%). In a cross-over study design, dolphins were instrumented and randomly assigned to AAV or CMV as the initial mode of ventilation, then switched to the alternate mode. Baseline cardiopulmonary data were collected and again after 30 min on each mode of ventilation. Cardiac index, stroke volume index, systemic vascular resistance, alveolar dead space, alveolar-arterial oxygen tension gradient, arterial oxygen content, oxygen delivery index, and dynamic respiratory system compliance index were calculated at each of the four time points. Results: During AAV, dolphins had higher arterial oxygen tension, higher mean airway pressure, reduced alveolar dead space ventilation and lower alveolar-arterial oxygen difference. Cardiovascular performance was not statistically different between the two modes. Discussion: Our study suggests AAV, which more closely resembles the conscious intermittent respiratory pattern phenotype of dolphins, improves ventilation and pulmonary function in the anesthetized dolphin. Future studies should evaluate the cardiopulmonary effects of neutral buoyancy and cardiopulmonary sparing drug protocols to reduce the need for hemodynamic support of current protocols.

The First Report of Pennella (Crustacea: Copepoda) Infesting Stenella coeruleoalba Stranded in Malta: Morphological and Genetic Analyses.

Here, we document the stranding of a striped dolphin Stenella coeruleoalba (Meyen, 1833) (Mammalia: Delphinidae), which was found dead in Maltese waters in July 2020. The stranded dolphin exhibited a severe infestation of the mesoparasitic copepod, Pennella balaenoptera Koren and Danielssen, 1877 (Copepoda: Pennelidae). Parasites of this genus represent the largest known mesoparasites to infest cetaceans. Under normal circumstances, cetaceans may have a few P. balaenoptera individuals attached to them, but cetaceans with compromised health are more prone to heavy infestations. The identification of the parasite was accomplished through morphological and genetic analyses. This incident highlights the significance of monitoring mesoparasitic infestations, offering valuable insights into the health of cetacean populations and emphasizing the potential implications for conservation efforts in the region.

Population distribution and drivers of habitat use for the Burrunan dolphins, Port Phillip Bay, Australia.

Bottlenose dolphin (Tursiops) populations, also described as the Burrunan dolphins, consist of a resident population of approximately 150 individuals in Port Phillip Bay (PPB), Victoria. Previous reports indicate distribution across a small southern region of PPB; however, little is known about their full distribution patterns across the entire PPB region. Here, we investigate the spatiotemporal distribution of the Burrunan dolphins across four zones representative of PPB benthic habitats and bathymetry to gain a better understanding of the potential drivers of the population's habitat use. Port Phillip Bay, Victoria, Australia. One hundred and twenty-nine boat-based surveys were undertaken between March 2015 and August 2021, encompassing 181 sightings. Generalised linear models (GLMs) were used to investigate annual, seasonal and zonal variation. We found no variation in sighting frequencies between years. Austral summer and winter had a significantly higher sighting frequency than autumn. We found that Burrunan dolphins utilise the entire bay, further extending the species range, and show a significantly higher number of sightings in the southern zone than in any other zones. Overlaying dolphin sightings with known oceanographic characteristics within PPB, we found bathymetry and benthic habitats were potential drivers for the Burrunan dolphins distribution and habitat use within the bay, with the dolphins significantly favouring the 5-10 and 10-15 m contour depths. These results show a more widespread distribution across the bay than previously documented. We recommend expansion of the current marine protected areas in the north and south of the bay. This study has increased our understanding of the vital habitat for the Burrunan dolphin populations. By providing evidence-based conservation recommendations, we hope to improve and contribute to future research, conservation management plans and effective marine protected areas across PPB for the resident Burrunan dolphin population.

Dietary exposure of potentially toxic elements to freshwater mammals in the Ganga river basin, India.

The threatened Gangetic dolphin (Platanista gangetica) and smooth-coated otter (Lutrogale perspicillata) of the Ganga River Basin (GRB), are experiencing a decline in their populations and distribution range owing to multiple anthropogenic pressures, including pollution by Potentially Toxic Elements (PTEs). Apex predators primarily encounter contaminants through dietary exposure. Yet, notable gaps persist in our understanding of the risks associated with the ingestion of PTE-contaminated prey for Gangetic Dolphins and Smooth-coated Otters. In this study, we examined the occurrence and spatial variation of PTEs in the prey fish of both these riverine mammals across three major rivers of GRB, while also evaluating the associated risk of ingesting contaminated prey. Our assessment revealed no statistical variation in bioaccumulation profiles of PTEs across the three rivers, attributable to comparable land use patterns and PTE consumption within the catchment. Zn and Cu were the most dominant PTEs in the prey species. The major potential sources of pollution identified in the catchment include agricultural settlements, vehicular emissions, and the presence of metal-based additives in plastics. Zn, As and Hg accumulation vary with the trophic level whereas some PTEs show concentration (Hg) and dilution (As, Cr, Pb and Zn) with fish growth. The Risk Quotient (RQ), based on the dietary intake of contaminated prey calculated using Toxicity Reference Value was consistently below 1 indicating no significant risk to these riverine mammals. Conversely, with the exception of Co and Ni, the Reference Dose-based RQs for all other PTEs indicated a substantial risk for Gangetic dolphins and smooth-coated otters through dietary exposure. This study serves as a pivotal first step in assessing the risk of PTEs for two threatened riverine mammals in a densely populated river basin, highlighting the importance of their prioritization in regular monitoring to reinforce the ongoing conservation efforts.

Novel processing and staining methodology of bottlenose dolphin (Tursiops truncatus) teeth for age determination.

Age determination of bottlenose dolphins (Tursiops truncatus) is a critical tool in understanding both individual and population health. There are many methods of aging bottlenose dolphins including analysis of teeth, pectoral flipper radiographs, and epigenetics. The most common and oldest method for aging toothed cetaceans is the counting of growth layer groups (GLGs) in the teeth. Current techniques have technical and repeatability challenges. Therefore, a processing technique that results in better resolution of GLGs is needed. This study compares different decalcifications and different histochemical staining techniques. Decalcification was done using 10% EDTA, Kristensen's decalcification, and Rapid Decalcification Solution (RDO). Following decalcification and routine processing, GLGs were assessed using Hematoxylin and Eosin (H&E), hematoxylin, Giemsa, Wright-Giemsa, Toluidine Blue (T-Blue), Masson's Trichrome, and Congo Red staining techniques. Decalcification with Kristensen's and staining with Masson's Trichrome and Congo Red were determined to best highlight GLGs. This processing and staining was then applied to a sample population of 102 bottlenose dolphins that were evaluated independently and blindly by two observers. Of the 102 dolphin samples, 13 (12.7%) were unable to age due to no clear distinction or distortion between GLGs.

Observational study on the non-linear response of dolphins to the presence of vessels.

With the large increase in human marine activity, our seas have become populated with vessels that can be overheard from distances of even 20 km. Prior investigations showed that such a dense presence of vessels impacts the behaviour of marine animals, and in particular dolphins. While previous explorations were based on a linear observation for changes in the features of dolphin whistles, in this work we examine non-linear responses of bottlenose dolphins (Tursiops Truncatus) to the presence of vessels. We explored the response of dolphins to vessels by continuously recording acoustic data using two long-term acoustic recorders deployed near a shipping lane and a dolphin habitat in Eilat, Israel. Using deep learning methods we detected a large number of 50,000 whistles, which were clustered to associate whistle traces and to characterize their features to discriminate vocalizations of dolphins: both structure and quantities. Using a non-linear classifier, the whistles were categorized into two classes representing the presence or absence of a nearby vessel. Although our database does not show linear observable change in the features of the whistles, we obtained true positive and true negative rates exceeding 90% accuracy on separate, left-out test sets. We argue that this success in classification serves as a statistical proof for a non-linear response of dolphins to the presence of vessels.

Accuracy of Dolphin Imaging software in predicting soft tissue changes of Class II Division 1 incisor relationship treated by all first premolar extraction compared with post-treatment lateral cephalogram.

BACKGROUND: To quantify the accuracy of Dolphin Imaging software in predicting soft tissue changes of Class II Division 1 incisor relationship treated by all first premolar extraction compared with actual changes. METHODS: Pretreatment and post-treatment lateral cephalograms of 30 young adult subjects (25 females and 5 males; age range of 16 to 21 years) were imported and traced in Dolphin Imaging software (version 11.95). Tracings of actual treatment results and the Dolphin predicted treatment outcome were superimposed to calculate the prediction errors. The mean difference for the Holdaway soft tissue analysis and at each landmark was measured in both the X and Y axes. Paired t test was used to compare the statistical differences in Holdaway soft tissue analysis parameters. RESULTS: There were significant differences between the predicted and actual values in three parameters of the Holdaway soft tissue analysis (P < 0.05). The overall accuracy of estimation was better in vertical direction than horizontal. Prediction of upper and lower lip landmarks was overestimated horizontally and underestimated vertically, whereas that belonging to chin was underestimated horizontally and overestimated vertically. Soft tissue nasion and soft tissue A-point were most accurately predicted, whereas the least accuracy was found in soft tissue chin region. CONCLUSIONS: Dolphin Imaging Software (version 11.95) can be accurate enough for use in patient communication and education but should be used with caution. It is most accurate in predicting soft tissue changes in the nasion and A-point regions, but least accurate in chin region.

First description of Rhodococcus equi infection in common bottlenose dolphin (Tursiops truncatus).

Rhodococcus equi is a terrestrial bacterium and a common pathogen in foals (Equus caballus), in which causes pneumonia. This report describes for the first time the infection caused by R. equi in a common bottlenose dolphin (Tursiops truncatus) stranded in the Calabrian coast, Italy. The post mortem examination of the animal revealed lesions in lung and colon. The animal was also positive to dolphin morbillivirus. The histological study showed lesions attributable to R. equi infection, such as pyogranulomatous bacterial pneumonia and chronic granulomatous colitis. Whole genome sequencing of the isolated strain confirmed its identification as R. equi.

Identification and characterization of plastic debris in the gastrointestinal tract of Guiana dolphins (Sotalia guianensis) from Espírito Santo coast, Brazil.

The Guiana dolphin (Sotalia guianensis) is categorized as vulnerable in the Brazilian list of endangered animals, and its populations suffer from several anthropological threats. In this study, we analyzed the presence of macro, meso, and microplastics (MPs) in Guiana dolphins (n = 12) in Brazil Southeastern coast by analysing their gastrointestinal tract. The MP extractions were carried out with H2O2 (35 %) to remove organic matter. Four specimens ingested meso and macroplastics, including an item of polypropylene of 19.22 cm that was produced about 943 km from the place in which the animal was found stranded. All the specimens analyzed had fragment-type microplastics in their intestines. Blue was the prevailing color, followed by black, green, and red. We highlight the contamination by microplastics in the species, still little investigated, especially the need to understand the contamination by microplastics along trophic levels.

Constrained spherical deconvolution on diffusion-weighted images of dolphin brains.

Invasive neuronal tract-tracing is not permitted in very large or endangered animals. This is especially the case in marine mammals like dolphins. Diffusion-weighted imaging of fiber tracts could be an alternative if feasible even in brains that have been fixed in formalin for a long time. This currently is a problem, especially for detecting crossing fibers. We applied a state-of-the-art algorithm of Diffusion-weighted imaging called Constrained Spherical Deconvolution on diffusion data of three fixed brains of bottlenose dolphins using clinical human MRI parameters and were able to identify complex fiber patterns within a voxel. Our findings indicate that in order to maintain the structural integrity of the tissue, short-term post-mortem fixation is necessary. Furthermore, pre-processing steps are essential to remove the classical Diffusion-weighted imaging artifacts from images: however, the algorithm is still able to resolve fiber tracking in regions with various signal intensities. The described imaging technique reveals complex fiber patterns in cetacean brains that have been preserved in formalin for extended periods of time and thus opens a new window into our understanding of cetacean neuroanatomy.

Effects of different initial speeds on subsequent glide and underwater undulatory swimming.

This study aimed to investigate the effect of different initial speeds on the performance during underwater undulatory swimming (UUS). The study included 13 female swimmers. Each participant was asked to perform a 15-m maximum UUS, starting with four different push-off speeds. The experiment was recorded using three underwater cameras; subsequently, a two-dimensional motion analysis was conducted. Statistical parametric mapping (SPM) was employed to identify the position where the UUS velocity stabilised. The findings revealed a significant difference in the average swimming velocities during the first cycle of UUS, which was attributed to the variation in initial speed (p < 0.05) while there is no significant difference in the middle and final cycles. The results of SPM analysis suggested that differences in UUS velocity became negligible after approximately 6-m position from the pool wall, regardless of variations in push-off velocity. Furthermore, it was confirmed that swimmers can reach their maximum achievable UUS velocity at approximately 5-m position, even if they fail to execute an effective push-off from the wall. These findings offer valuable insights for future UUS studies, specifically in choosing suitable cycles for analysis.

The Ecology of Non-Candida Yeasts and Dimorphic Fungi in Cetaceans: From Pathogenicity to Environmental and Global Health Implications.

Cetaceans, which are integral to marine ecosystems, face escalating anthropogenic threats, including climate change and pollution, positioning them as critical sentinel species for ocean and human health. This review explores the neglected realm of non-Candida yeasts in cetaceans, addressing the gaps in the understanding of their prevalence, pathogenicity, and environmental impacts. By examining identified species such as Cryptococcus spp., Paracoccidioides spp., and several dimorphic fungi, this review emphasizes global prevalence, epidemiology and ecology, pathogenicity, and potential zoonotic implications. It also discusses the fine line between yeast commensalism and pathogenicity by considering environmental influences such as pollution, climate shifts, and immune suppression. Environmental impact discussions delve into how rising ocean temperatures and pollution can modify yeast mycobiota, potentially affecting marine host health and broader ecosystem dynamics. The cetacean's unique physiology and ecological niches are considered, highlighting potential impacts on behaviors, reproductive success, and survival rates. Identifying crucial knowledge gaps, the review calls for intensified research efforts, employing advanced molecular techniques to unravel the cetacean mycobiome. Systematic studies on yeast diversity, antifungal susceptibility, and their influence on environmental and ecosystem health are proposed, and the balance between commensal and pathogenic species emphasizes the significance of the One Health approach. In conclusion, as marine mammals face unprecedented challenges, unveiling non-Candida yeasts in cetaceans emerges as a critical endeavor with far-reaching implications for the conservation of marine ecosystems and for both animal and human public health.

Distribution of calcium-binding proteins immunoreactivity in the bottlenose dolphin entorhinal cortex.

Introduction: The entorhinal cortex has been shown to be involved in high-level cognitive functions in terrestrial mammals. It can be divided into two main areas: the lateral entorhinal area (LEA) and the medial entorhinal area (MEA). Understanding of its structural organization in cetaceans is particularly important given the extensive evidence for their cognitive abilities. The present study describes the cytoarchitectural and immunohistochemical properties of the entorhinal cortex of the bottlenose dolphin (Tursiops truncatus, Montagu, 1821), perhaps the most studied cetacean species and a paradigm for dolphins and other small cetaceans. Methods: Four bottlenose dolphins' entorhinal cortices were processed. To obtain a precise overview of the organization of the entorhinal cortex we used thionin staining to study its laminar and regional organization, and immunoperoxidase technique to investigate the immunohistochemical distribution of three most commonly used calcium-binding proteins (CBPs), calbindin D-28k (CB), calretinin (CR) and parvalbumin (PV). Entorhinal cortex layers thickness were measured, morphological and morphometric analysis for each layer were conducted and statistically compared. Results: Six layers in both the LEA and MEA were identified. The main difference between the LEA and the MEA is observed in layers II and III: the neurons in layer II of the LEA were denser and larger than the neurons in layer II of MEA. In addition, a relatively cell-free zone between layers II and III in LEA, but not in MEA, was observed. The immunohistochemical distribution of the three CBPs, CB, CR and PV were distinct in each layer. The immunostaining pattern of CR, on one side, and CB/PV, on the other side, appeared to be distributed in a complementary manner. PV and CB immunostaining was particularly evident in layers II and III, whereas CR immunoreactive neurons were distributed throughout all layers, especially in layers V and VI. Immunoreactivity was expressed by neurons belonging to different morphological classes: All CBPs were expressed in non-pyramidal neurons, but CB and CR were also found in pyramidal neurons. Discussion: The morphological characteristics of pyramidal and non-pyramidal neurons in the dolphin entorhinal cortex are similar to those described in the entorhinal cortex of other species, including primates and rodents. Interestingly, in primates, rodents, and dolphins, most of the CBP-containing neurons are found in the superficial layers, but the large CR-ir neurons are also abundant in the deep layers. Layers II and III of the entorhinal cortex contain neurons that give rise to the perforant pathway, which conveys most of the cortical information to the hippocampal formation. From the hippocampal formation, reciprocal projections are directed back to the deep layer of the entorhinal cortex, which distributes the information to the neocortex and subcortical area. Our data reveal that in the dolphin entorhinal cortex, the three major CBPs label morphologically heterogeneous groups of neurons that may be involved in the information flow between entorhinal input and output pathways.

Broad-scale impacts of coastal mega-infrastructure project on obligatory inshore delphinids: A cautionary tale from Hong Kong.

Inshore marine habitats experience considerable anthropogenic pressure, as this is where many adverse effects of human activities concentrate. In the rapidly-changing seascape of the Anthropocene, Hong Kong waters at the heart of world's fastest developing coastal region can serve as a preview-window into coastal seas of the future, with ever-growing anthropogenic footprint. Here, we quantify how large-scale coastal infrastructure projects can affect obligatory inshore cetaceans, bringing about population-level consequences that may compromise their long-term demographic viability. As a case in point, we look at the construction of world's longest sea crossing system and broad-scale demographic, social and spatial responses it has caused in a shallow-water delphinid, the Indo-Pacific humpback dolphin (Sousa chinensis). Soon after the infrastructure project began, dolphins markedly altered their home range near construction sites such that these waters no longer functioned as dolphin core areas despite the apparent presence of prey, indicating that anthropogenic impacts outweighed foraging benefits. The contraction of key habitats has in turn led individuals to interact over spatially more constricted area, reshaping their group dynamics and social network. Although there was no apparent decline in dolphin numbers that could be detected with mark-recapture estimates, adult survival rates decreased drastically from 0.960 to 0.904, the lowest estimate for these animals anywhere across the region to date, notably below the previously estimated demographic threshold of their long-term persistence (0.955). It is apparent that during an advanced stage of this coastal infrastructure project, dolphins were under a major anthropogenic pressure that, if sustained, could be detrimental to their long-term persistence as a viable demographic unit. As effective conservation of species and habitats depends on informed management decisions, this study offers a valuable lesson in environmental risk assessment, underscoring the implications of human-induced rapid environmental change on obligatory inshore delphinids-sentinels of coastal habitats that are increasingly degraded in fast-changing coastal seas.

Vascularization of the gastrointestinal tract of the bottlenose dolphin (Tursiops truncatus, Montagu 1821).

Odontocetes primarily rely on fish, cephalopods, and crustaceans as their main source of nutrition. In the digestive system, their polygastric complex exhibits similarities to that of their closest terrestrial relatives such as cows, sheep, and giraffes, while the entero-colic tract shares similarities with terrestrial carnivores. The morphology, caliber, and structure of the odontocete intestine are relatively constant, and, since there is no caecum, a distinction between the small and large intestine and their respective subdivisions is difficult. To address this issue, we used the intestinal vascularization pattern, specifically the course and branching of the celiac artery (CA) and the cranial and caudal mesenteric arteries (CrMA and CdMA). A series of pictures and dissections of 10 bottlenose dolphins (Tursiops truncatus) were analyzed. Additionally, we performed a cast by injecting colored polyurethane foam in both arteries and veins to measure the caliber of the arteries and clarify their monopodial or dichotomous branching. Our results showed the presence of multiple duodenal arteries (DAs) detaching from the CA. The CrMA gave origin to multiple jejunal arteries, an ileocolic artery (ICA), and, in six cases, a CdMA. In four specimens, the CdMA directly originated from the abdominal aorta. The ICA gave rise to the mesenteric ileal branches (MIB) and mesenteric anti-ileal branches and the right colic arteries (RCA) and the middle colic arteries. From the CdMA originated the left colic and cranial rectal arteries (LCA and CrRA). The measurements revealed a mixed monopodial and dichotomous branching scheme. The analysis of the arteries and their branching gave us an instrument, based on comparative anatomy, to distinguish between the different intestinal compartments. We used the midpoint of anastomoses between MIB and RCA to indicate the border between the small and the large intestine, and the midpoint of anastomoses between LCA and CrRA, to tell the colon from the rectum. This pattern suggested an elongation of the duodenum and a shortening of the colic tract that is still present in this species. These findings might be related to the crucial need to possess a long duodenal tract to digest prey ingested whole without chewing. A short aboral part is also functional to avoid gas-producing colic fermentation. The rare origin of the CdMA on the CrMA might instead be a consequence of the cranial thrust of the abdominopelvic organs related to the loss of the pelvic girdle that occurred during the evolution of cetaceans.

Brucella infection in rough-toothed dolphin (Steno bredanensis) with severe orchitis stranded on the Pacific coast of Japan.

In the western North Pacific, prominent granulomatous testes have been detected in many Brucella-infected common minke whales (Balaenoptera acutorostrata), but there have been no reports in toothed cetaceans. We found severe orchitis with granulomatous lesions in a rough-toothed dolphin (Steno bredanensis) stranded on the Pacific coast of Japan in 2011. Histopathological examination revealed leukocyte infiltration of the lesions. DNA from the lesion was analyzed by PCR and it showed molecular biological similarities with those of Brucella-infected common minke whales and Brucella ceti of sequence-type 27 (ST27). These results suggest that the type of Brucella ceti that infected the dolphin was ST27, which may have caused severe orchitis. This study adds to our understanding of Brucella infections in marine mammals.

Optimization and validation of a micro-QuEChERS method for phthalates detection in small samples of cetacean blubber.

In this study, an innovative method was developed to detect and quantify phthalates in fresh cetacean blubber. An adaptation of the ammonium formate QuEChERS method was used and adapted as a micro-extraction for small quantities of samples. Significantly, this technique utilized minimal quantities of reagents and salts, with the additional implementation of rigorous Quality Assurance/Quality Control protocols to further reduce background contamination. To ensure the reliability of this method, comprehensive validation procedures were conducted, with a specific focus on two widely studied cetacean species: the common bottlenose dolphin (Tursiops truncatus) and the short-finned pilot whale (Globicephala macrorhynchus). Determination coefficients (R2) for matrix-matched calibration were >0.93 with limits of quantifications (LOQ) of the method in the range of 5-10 ng/g. Mean recovery values were between 40 and 100 %. This novel methodology holds particular relevance for environmental research studies, offering the capability to detect emerging contaminants with minimal sample requirements. This aspect is particularly valuable in investigations that involve free-ranging animals and rely on biopsy sampling. It allows for the assessment of contaminant levels in healthy individuals within wild populations, enhancing our understanding of ecological impacts and potential conservation measures.•A micro-extraction adaptation of the ammonium formate QuEChERS method was developed and applied to a small quantity of fresh cetacean blubber to detect phthalates.•Small quantities of reagents and salts were used, and additional Quality Assurance/ Quality Control procedures were taken to further minimize background contamination.•Method validation was carried out for two cosmopolitan and extensively studied cetacean species: the common bottlenose dolphin (Tursiops truncatus) and the short-finned pilot whale (Globicephala macrorhynchus).

Survey of exposure to stranded dolphins in Japan to investigate an outbreak of suspected infection with highly pathogenic avian influenza (H5N1) clade 2.3.4.4(b) in humans.

Background: A highly pathogenic avian influenza (HPAI) A (H5N1) virus has been detected in domestic and wild animals worldwide. The incidence of HPAI infections in sea mammals has been increasing, as is the number of stranded marine mammals linked to H5N1 viral clade 2.3.4.4(b). In this study, we investigated a stranding event involving dolphins and human-dolphin contact, and investigated the potential risk of animal-to-human H5N1 transmission with a survey of exposure on the Tsurigasaki coast, Japan. Methods: We performed a non-random, convenient-sample-based, survey on Tsurigasaki beach where around 30 melon-headed whales were stranded on April 3, 2023. Face-to-face (n = 25) and telephone (n = 1) interviews among surfers took place on April 7 and 8. A nasal swab for quick antigen testing was taken from those who wished to be tested (n = 13), to detect infections with influenza A virus. Results: Although there was no confirmatory diagnosis of H5N1 in either humans or dolphins (while n = 3 dolphins were autopsied), we found that a large number of surfers had touched the dolphins with their bare hands while attempting to rescue them, and that some surfers were directly exposed to dolphin blood and body fluids in the ocean. Conclusions: The adequate communication of risk is required to minimize the threat of viral transmission at this particular human-animal interface. Administrative and legal responses to cross-species transmission, including guidelines via one health frameworks, a rapid evaluation process of ethical approval, and the systematic involvement of experts in infectious disease, must be urgently formulated.

Pre-anal gland abscess in a male Atlantic bottlenose dolphin (Tursiops truncatus).

OBJECTIVE: Anal glands have been identified in a variety of terrestrial and aquatic mammalian species, but there are few accounts describing their presence in cetaceans. To our knowledge, this report describes the first documented case of a pre-anal gland abscess in an Atlantic bottlenose dolphin (Tursiops truncatus). ANIMAL: A 9-year-old male bottlenose dolphin (T truncatus) part of the US Navy Marine Mammal Program in San Diego Bay, California. CLINICAL PRESENTATION, PROGRESSION, AND PROCEDURES: The patient presented for a 3-day history of lethargy, failure to perform voluntary behaviors, and an elevated respiratory rate. Complete blood count and serum biochemistry results showed an inflammatory hemogram. Physical examination revealed a 4-cm circular swelling at the right pre-anal gland pore. The swelling was warm and erythematous, with multifocal pinpoint ulcerations. An abscess of the pre-anal gland was diagnosed using cytology, culture, and ultrasound. TREATMENT AND OUTCOME: Treatment included systemic oral antibiotic and antifungal therapy, along with daily lavage and warm compress of the gland. Treatment was successful, and the abscess resolved. CLINICAL RELEVANCE: This case provides insight into a previously unreported disease process in bottlenose dolphins and encourages veterinarians to evaluate the pre-anal gland during routine physical examinations and complete further work-up if swelling or clinical signs associated with this region are present.