Cook Inlet beluga whale Delphinapterus leucas with valvular endocarditis, encephalitis, rhabdomyolysis, heavy parasitism and fungal dermatitis.

This is a case report of a Cook Inlet beluga whale Delphinapterus leucas found dead stranded on September 28, 2020 in Turnagain Arm, Alaska. This subadult male had valvular endocarditis, encephalitis, rhabdomyolysis, myoglobinuric nephropathy, severe parasitism and fungal dermatitis. Erysipelothrix rhusiopathiae was detected in the heart lesion, eye and external swabs. The level of infection and parasitism in this individual is markedly higher than what has been found in other Cook Inlet belugas, suggesting immunosuppression.

Serological Detection of Marine Origin Brucella Exposure in Two Alaska Beluga Stocks.

Among emerging threats to the Arctic is the introduction, spread, or resurgence of disease. Marine brucellosis is an emerging disease concern among free-ranging cetaceans and is less well-studied than terrestrial forms. To investigate marine-origin Brucella sp. exposure in two beluga stocks in Alaska, USA, this study used serological status as well as real-time polymerase chain reaction (rtPCR) and bacterial culture. In total, 55 live-captured-released belugas were tested for Brucella exposure in Bristol Bay (2008-2016) and 112 (8 live-captured; 104 subsistence-harvested) whales were tested in the eastern Chukchi Sea (2007-2017). In total, 73% percent of Bristol Bay live captures, 50% of Chukchi Sea live captures, and 66% of Chukchi Sea harvested belugas were positive on serology. Only 10 of 69 seropositive belugas were rtPCR positive in at least one tissue. Only one seropositive animal was PCR positive in both the spleen and mesenteric lymph node. All animals tested were culture negative. The high prevalence of seropositivity detected suggests widespread exposure in both stocks, however, the low level of rtPCR and culture positive results suggests clinical brucellosis was not prevalent in the belugas surveyed. Continued detection of Brucella exposure supports the need for long-term monitoring of these and other beluga populations.

A Virtual Necropsy: Applications of 3D Scanning for Marine Mammal Pathology and Education.

Stranded large whales represent an opportunity to learn about the anatomy and health of these cryptic free-ranging animals. However, where time and access is frequently limited, law enforcement and management priorities often take precedence over research, outreach, and educational uses. On 14 March 2021, a dead female adult humpback whale was reported stranded on an uninhabited island 15 miles west of Sitka, Alaska. The whale was three-dimensionally scanned using light detection and ranging (LiDAR) and photogrammetry before, during, and at multiple time points after a necropsy, including full decomposition 17 days later (NOAA Fisheries permit 18786-01). These scans were organized and displayed on the site Sketchfab with annotations and made publically available as a "4D virtual necropsy" (the fourth dimension is time). After one month, our user survey indicated widespread interest in the platform by both the local community and worldwide by stranding professionals, researchers, and educators. We are unaware of another 3D scan involving a large whale with soft tissue for teaching, research, or public display, despite the ease of 3D scanning with current technologies and the wide-ranging applications.

Adenovirus Hemorrhagic Disease in Moose (Alces americanus gigas) in Alaska, USA.

In 1993, an epizootic of adenovirus hemorrhagic disease (AHD) caused the death of at least 1,000 mule deer (Odocoileus hemionus) in California, US. Since then, numerous cervid species throughout the US have had deaths confirmed to be caused by AHD. In 2015, the death of two captive moose (Alces americanus gigas) calves marked the first recognized AHD-caused deaths in Alaska, a state in which moose are important economically as well as for food security and cultural identity. Both cases were characterized by systemic vasculitis with endothelial cell intranuclear inclusion bodies, pulmonary edema, petechial hemorrhages, and enterotyphlocolitis.

Pathology findings and correlation with body condition index in stranded killer whales (Orcinus orca) in the northeastern Pacific and Hawaii from 2004 to 2013.

Understanding health and mortality in killer whales (Orcinus orca) is crucial for management and conservation actions. We reviewed pathology reports from 53 animals that stranded in the eastern Pacific Ocean and Hawaii between 2004 and 2013 and used data from 35 animals that stranded from 2001 to 2017 to assess association with morphometrics, blubber thickness, body condition and cause of death. Of the 53 cases, cause of death was determined for 22 (42%) and nine additional animals demonstrated findings of significant importance for population health. Causes of calf mortalities included infectious disease, nutritional, and congenital malformations. Mortalities in sub-adults were due to trauma, malnutrition, and infectious disease and in adults due to bacterial infections, emaciation and blunt force trauma. Death related to human interaction was found in every age class. Important incidental findings included concurrent sarcocystosis and toxoplasmosis, uterine leiomyoma, vertebral periosteal proliferations, cookiecutter shark (Isistius sp.) bite wounds, excessive tooth wear and an ingested fish hook. Blubber thickness increased significantly with body length (all p < 0.001). In contrast, there was no relationship between body length and an index of body condition (BCI). BCI was higher in animals that died from trauma. This study establishes a baseline for understanding health, nutritional status and causes of mortality in stranded killer whales. Given the evidence of direct human interactions on all age classes, in order to be most successful recovery efforts should address the threat of human interactions, especially for small endangered groups of killer whales that occur in close proximity to large human populations, interact with recreational and commercial fishers and transit established shipping lanes.

Genomic Characterization of Picornaviruses Isolated From Ribbon (Histriophoca fasciata) and Harbor (Phoca vitulina) Seals.

The seal picornavirus 1, species Aquamavirus A, is currently the only recognized member of the genus Aquamavirus within the family Picornaviridae. The bear picornavirus 1 was recently proposed as the second species in the genus under the name aquamavirus B. Herein, we determined the complete genomes of two novel pinniped picornaviruses, the harbor seal picornavirus (HsPV) and the ribbon seal picornavirus (RsPV). The HsPV and the RsPV were isolated in Vero.DogSLAMtag cells from samples collected from stranded harbor (Phoca vitulina) and ribbon (Histriophoca fasciata) seals. RsPV-infected Vero.DogSLAMtag cells displaying extensive cytopathic effects were processed for transmission electron microscopy and revealed non-enveloped viral particles aggregated into paracrystalline arrays in the cytoplasm. A next-generation sequencing approach was used to recover the complete genomes of the HsPV and the RsPV (6,709 and 6,683 bp, respectively). Phylogenetic and genetic analyses supported the HsPV and the RsPV as members of the Aquamavirus genus. Based on these results, RsPV represents a novel strain of Aquamavirus A, while the HsPV is a novel strain of the proposed species aquamavirus B. These discoveries provide information on the evolutionary relationships and ultrastructure of aquamaviruses and expands the known host range of those viruses. Our results underscore the importance of the application of classical virology and pathology techniques coupled with high-throughput sequencing technologies for the discovery and characterization of pathogens in wild marine mammals.

Skin microbiome of beluga whales: spatial, temporal, and health-related dynamics.

BACKGROUND: Host-specific microbiomes play an important role in individual health and ecology; in marine mammals, epidermal microbiomes may be a protective barrier between the host and its aqueous environment. Understanding these epidermal-associated microbial communities, and their ecological- or health-driven variability, is the first step toward developing health indices for rapid assessment of individual or population health. In Cook Inlet, Alaska, an endangered population of beluga whales (Delphinapterus leucas) numbers fewer than 300 animals and continues to decline, despite more than a decade of conservation effort. Characterizing the epidermal microbiome of this species could provide insight into the ecology and health of this endangered population and allow the development of minimally invasive health indicators based on tissue samples. RESULTS: We sequenced the hypervariable IV region of bacterial and archaeal SSU rRNA genes from epidermal tissue samples collected from endangered Cook Inlet beluga whales (n = 33) and the nearest neighboring population in Bristol Bay (n = 39) between 2012 and 2018. We examined the sequences using amplicon sequence variant (ASV)-based analyses, and no ASVs were associated with all individuals, indicating a greater degree of epidermal microbiome variability among beluga whales than in previously studied cetacean species and suggesting the absence of a species-specific core microbiome. Epidermal microbiome composition differed significantly between populations and across sampling years. Comparing the microbiomes of Bristol Bay individuals of known health status revealed 11 ASVs associated with potential pathogens that differed in abundance between healthy individuals and those with skin lesions or dermatitis. Molting and non-molting individuals also differed significantly in microbial diversity and the abundance of potential pathogen-associated ASVs, indicating the importance of molting in maintaining skin health. CONCLUSIONS: We provide novel insights into the dynamics of Alaskan beluga whale epidermal microbial communities. A core epidermal microbiome was not identified across all animals. We characterize microbial dynamics related to population, sampling year and health state including level of skin molting. The results of this study provide a basis for future work to understand the role of the skin microbiome in beluga whale health and to develop health indices for management of the endangered Cook Inlet beluga whales, and cetaceans more broadly.

Comparing progesterone in blubber and serum to assess pregnancy in wild beluga whales (Delphinapterus leucas).

The Cook Inlet population of beluga whales (Delphinapterus leucas) is listed as endangered and continues to decline for largely unknown reasons; however, there is some evidence that poor reproductive success is a contributing factor. Pregnancy is difficult to detect through observation, and, there is reluctance to capture endangered beluga whales for reproductive tract imaging via ultrasound or to obtain suitable samples for pregnancy assessments. An endocrine analysis of blubber biopsies collected by remote darting could represent a minimally invasive way to identify pregnant females and compare pregnancy rates among years or populations. Studies have validated the use of blubber biopsies to identify pregnant females in other cetacean species, but not beluga whales; therefore, validation of blubber progesterone levels to proven tests that reliably detect pregnancy was needed for this species. As part of a larger study, we sampled blood and blubber from live-captured beluga whales (21 females, 26 males) in Bristol Bay, Alaska. Progesterone levels were determined in serum samples obtained from all animals and in blubber samples from a subset (14 females, 13 males) to determine pregnancy status, estimate the stage of pregnancy, and evaluate the suitability of using blubber alone for these assessments. In general, there was distinct separation of high levels of progesterone in serum and blubber for presumed pregnant females and low levels for males and presumed non-pregnant females. Blubber progesterone levels in two females (14% of females tested) were intermediate (i.e. ambiguous); their corresponding serum levels were consistent with being pregnant in one case and not being pregnant in the other. Except for these two intermediate values, pregnancy status of beluga whales could be determined from blubber alone, thereby providing a valuable tool to better understand reproduction dynamics from populations that cannot be captured for examination.

Characterization of a novel rhabdovirus isolated from a stranded harbour porpoise (Phocoena phocoena).

An adult male harbour porpoise (Phocoena phocoena) stranded off the coast of Alaska displaying poor body condition, scattered mild ulcerative dermatitis, and necrotizing balanoposthitis. Necropsy findings included severe verminous panniculitis, pneumonia, hepatitis, and enteritis. Histopathological examination of skin lesions revealed a pustular epidermitis and dermatitis, with ballooning degeneration of keratinocytes and occasional amphophilic intracytoplasmic inclusion bodies. A swab sample collected from the ulcerative penile lesions was processed for virus isolation resulting in cytopathic effects observed in primary beluga whale kidney (BWK) cells. Transmission electron microscopy revealed bullet-shaped virions budding from the cell surface of infected BWK cells consistent with a rhabdovirus. A cDNA library was prepared using RNA extracted from infected cell culture supernatant and sequenced on an Illumina MiSeq sequencer. The near-complete genome of a novel rhabdovirus was recovered. Genetic and phylogenetic analyses based on the complete L gene supported the harbour porpoise rhabdovirus (HPRV) as a new species. HPRV clustered phylogenetically with dolphin rhabdovirus (DRV) and this cetacean rhabdovirus clade was found to be the sister group to members of the genus Perhabdovirus that infect fish. A specific nested RT-PCR assay detected HPRV RNA in the epaxial musculature of the harbour porpoise. Our results are consistent with a previous hypothesis that cetacean rhabdoviruses may have arisen following a host jump from fish and suggest that DRV and HPRV represent separate species belonging in a new genus within the family Rhabdoviridae. Further research is needed to determine the health impact of HPRV in harbour porpoise populations, its prevalence, and route of transmission.

Concentrations of organohalogens (PCBs, DDTs, PBDEs) in hunted and stranded Northern sea otters (Enhydra lutris kenyoni) in Alaska from 1992 to 2010: Links to pathology and feeding ecology.

Many organohalogen compounds (OHCs) are persistent organic pollutants (POPs) found in appreciable concentrations in marine predators. While production of some POPs has declined or ceased in recent decades, their capacity for global transport and bioaccumulation results in observations of unchanging or increasing concentrations in marine systems. Sea otters (Enhydra lutris) have been advocated as an environmental sentinel for contaminants due to their longevity, site fidelity and prey species that often overlap with human consumption. Using archived (1992-2010) samples of livers from Northern sea otters (n = 50) from Alaska we examine concentrations of chlordanes (CHLs), polychlorinated biphenyls (PCBs), dichlorodiphenyltrichloroethane (DDTs), and polybrominated diphenyl ethers (PBDEs) and associated metabolites. We found some evidence for declining ΣPCBs over the two decades, however for most animals concentrations were low compared to toxicological thresholds. Six animals had relatively high concentrations of ΣPCBs (mean = 262,000 ng/g lipid weight), ΣDDTs (mean = 8,800 ng/g lw), and ΣPBDEs (mean = 4,600 ng/g lw), with four of these six animals experiencing hepatic parasitism or hepatitis. In order to assess whether differences in POP concentrations are associated with feeding ecology, we examined stable isotopes of C and N in archived muscle and whisker samples. In general, there were no significant relationships between ΣPOP concentrations and stable isotope ratios. There were small differences in stable isotope profiles in animals with high POP concentrations, although it was unclear if these differences were due to feeding ecology or disease processes. This study highlights the importance of considering feeding ecology and necropsy (health and disease status) data while conducting contaminant surveys, and confirms some previous reports of trends in OHCs in Alaska marine mammals.

Oil fouling in three subsistence-harvested ringed (Phoca hispida) and spotted seals (Phoca largha) from the Bering Strait region, Alaska: Polycyclic aromatic hydrocarbon bile and tissue levels and pathological findings.

Oil spills of unknown origin were detected in three oil-fouled, ice-associated seals from the Alaska Bering Strait region collected by Alaska Native subsistence hunters during fall 2012. Bile analyses of two oiled seals indicated exposure to fluorescent polycyclic aromatic hydrocarbon (PAH) metabolites but levels of some metabolites were similar to or lower than biliary levels in harvested unoiled ice seals. Oiled seals had elevated tissue PAH concentrations compared to tissue levels of PAHs determined in unoiled ice seals. However, regardless of oiling status, tissue PAH levels were relatively low (<50 ng/g, wet weight) likely due to rapid PAH metabolism and elimination demonstrated previously by vertebrates. Hepatic, pulmonary, and cardiac lesions were observed in oiled seals in conjunction with measurable PAHs in their tissue and bile. This is the first study to report tissue and bile PAH concentrations and pathologic findings of oiled ice seals from the U.S. Arctic.

Genome Sequence of an Alphaherpesvirus from a Beluga Whale (Delphinapterus leucas).

Beluga whale alphaherpesvirus 1 was isolated from a blowhole swab taken from a juvenile beluga whale. The genome is 144,144 bp in size and contains 86 putative genes. The virus groups phylogenetically with members of the genus Varicellovirus in subfamily Alphaherpesvirinae and is the first alphaherpesvirus sequenced from a marine mammal.

De novo transcriptome assembly and RNA-Seq expression analysis in blood from beluga whales of Bristol Bay, AK.

Assessing the health of marine mammal sentinel species is crucial to understanding the impacts of environmental perturbations on marine ecosystems and human health. In Arctic regions, beluga whales, Delphinapterus leucas, are upper level predators that may serve as a sentinel species, potentially forecasting impacts on human health. While gene expression profiling from blood transcriptomes has widely been used to assess health status and environmental exposures in human and veterinary medicine, its use in wildlife has been limited due to the lack of available genomes and baseline data. To this end we constructed the first beluga whale blood transcriptome de novo from samples collected during annual health assessments of the healthy Bristol Bay, AK stock during 2012-2014 to establish baseline information on the content and variation of the beluga whale blood transcriptome. The Trinity transcriptome assembly from beluga was comprised of 91,325 transcripts that represented a wide array of cellular functions and processes and was extremely similar in content to the blood transcriptome of another cetacean, the bottlenose dolphin. Expression of hemoglobin transcripts was much lower in beluga (25.6% of TPM, transcripts per million) than has been observed in many other mammals. A T12A amino acid substitution in the HBB sequence of beluga whales, but not bottlenose dolphins, was identified and may play a role in low temperature adaptation. The beluga blood transcriptome was extremely stable between sex and year, with no apparent clustering of samples by principle components analysis and <4% of genes differentially expressed (EBseq, FDR<0.05). While the impacts of season, sexual maturity, disease, and geography on the beluga blood transcriptome must be established, the presence of transcripts involved in stress, detoxification, and immune functions indicate that blood gene expression analyses may provide information on health status and exposure. This study provides a wealth of transcriptomic data on beluga whales and provides a sizeable pool of preliminary data for comparison with other studies in beluga whale.

Coxiella burnetii exposure in northern sea otters Enhydra lutris kenyoni.

Valvular endocarditis has been well described in northern sea otters Enhydra lutris kenyoni of Alaska and in many cases no cause has been identified. It is also one of the most common conditions observed in people with chronic Coxiella burnetii infection. Given the high levels of C. burnetii exposure in marine mammals distributed throughout the same geographic range as the northern sea otter, and the presence of valvular lesions seen in otters, the objective of this study was to determine the level of C. burnetii exposure in otters and investigate any association between exposure, infection and valvular disease in this species. Archived serum from 75 live captured, apparently healthy otters (25 from each of 3 stocks) and 30 dead otters were tested for C. burnetii antibodies by indirect florescent antibody assay (IFA). Archived bone marrow and heart valves were tested for C. burnetii DNA by real-time PCR (qPCR). Overall, the seroprevalence in live otters was 17%, with significantly more exposed animals in the south central (40%) stock relative to the southwest (8%) and southeast (4%). The seroprevalence of animals sampled post mortem was 27%, although none of the bone marrow or heart valve samples were positive by qPCR. Results of this study failed to demonstrate a significant association between C. burnetii infection and valvular endocarditis in sea otters; however, the differing seroprevalence suggests that exposure opportunities vary geographically.

Prevalence and spatio-temporal variation of an alopecia syndrome in polar bears (Ursus maritimus) of the southern Beaufort Sea.

Alopecia (hair loss) has been observed in several marine mammal species and has potential energetic consequences for sustaining a normal core body temperature, especially for Arctic marine mammals routinely exposed to harsh environmental conditions. Polar bears (Ursus maritimus) rely on a thick layer of adipose tissue and a dense pelage to ameliorate convective heat loss while moving between sea ice and open water. From 1998 to 2012, we observed an alopecia syndrome in polar bears from the southern Beaufort Sea of Alaska that presented as bilaterally asymmetrical loss of guard hairs and thinning of the undercoat around the head, neck, and shoulders, which, in severe cases, was accompanied by exudation and crusted skin lesions. Alopecia was observed in 49 (3.45%) of the bears sampled during 1,421 captures, and the apparent prevalence varied by years with peaks occurring in 1999 (16%) and 2012 (28%). The probability that a bear had alopecia was greatest for subadults and for bears captured in the Prudhoe Bay region, and alopecic individuals had a lower body condition score than unaffected individuals. The cause of the syndrome remains unknown and future work should focus on identifying the causative agent and potential effects on population vital rates.

Sarcocystis canis associated hepatitis in a Steller sea lion (Eumetopias jubatus) from Alaska.

Sarcocystis canis infection was associated with hepatitis in a Steller sea lion (Eumetopias jubatus). Intrahepatocellular protozoal schizonts were among areas of necrosis and inflammation. The parasite was genetically identical to S. canis and is the first report in a Steller sea lion, indicating another intermediate host species for S. canis.

Locally acquired disseminated histoplasmosis in a northern sea otter (Enhydra lutris kenyoni) in Alaska, USA.

Histoplasmosis of local origin has not been reported in humans or wildlife in Alaska, and the disease has never been reported in a free-ranging marine mammal. In 2005 a northern sea otter (Enhydra lutris kenyoni) was found on Kodiak Island, Alaska, at 57° latitude north, far outside the known distribution of Histoplasma capsulatum. The animal died of disseminated histoplasmosis. Microorganisms consistent with Histoplasma sp. were observed on histopathology, and H. capsulatum was identified by PCR and sequencing. We suggest migratory seabirds or aerosol transmission through prevailing winds may have resulted in transmission to the sea otter.

Sarcomas in three free-ranging northern sea otters (Enhydra lutris kenyoni) in Alaska.

Three sarcomas were diagnosed in wild northern sea otters (Enhydra lutris kenyoni) during the mid- to late 1990s. Histologically, the tumors were a chondrosarcoma and two low-grade fibrosarcomas with myofibroblastic cell differentiation. The three sea otters were surviving in the wild and were killed by hunters.