Hepatitis E Virus Infections in Free-Ranging and Captive Cetaceans, Spain, 2011-2022.

Epidemiologic surveillance of hepatitis E virus in over 300 free-ranging and captive cetaceans in waters off Spain revealed extensive exposure to this pathogen. We suggest the persistent and widespread presence of hepatitis E in the marine environment off the coast of Spain may be driven by terrestrial sources of contamination.

Microscopic Findings in the Cardiac Muscle of Stranded Extreme Deep-Diving Cuvier's Beaked Whales (Ziphius cavirostris).

Considerable information has been gained over the last few decades on several disease processes afflicting free-ranging cetaceans from a pathologist's point of view. Nonetheless, there is still a dearth of studies on the hearts of these species. For this reason, we aimed to improve our understanding of cardiac histological lesions occurring in free-ranging stranded cetaceans and, more specifically, in deep-diving Cuvier's beaked whales. The primary cardiac lesions that have been described include vascular changes, such as congestion, edema, hemorrhage, leukocytosis, and intravascular coagulation; acute degenerative changes, which consist of contraction band necrosis, wavy fibers, cytoplasmic hypereosinophilia, and perinuclear vacuolization; infiltration of inflammatory cells; and finally, the presence and/or deposition of different substances, such as interstitial myoglobin globules, lipofuscin pigment, polysaccharide complexes, and intra- and/or extravascular gas emboli and vessel dilation. This study advances our current knowledge about the histopathological findings in the cardiac muscle of cetaceans, and more specifically, of Cuvier's beaked whales.

Retrospective immunohistochemical investigation on dolphin morbillivirus infection by comparing the performance of heterologous monoclonal and polyclonal antibodies - Short communication.

Dolphin morbillivirus (DMV) is a pathogen of great concern in free-ranging cetaceans. Confirmation and staging of morbillivirus infections rely on histology and immunohistochemistry (IHC), following molecular detection. As at the present time no specific antibodies (Abs) against DMV are available, two heterologous Abs have been used worldwide for the examinations of morbillivirus infections of cetaceans. One is a monoclonal Ab (MoAb) prepared against the N protein of canine distemper virus (CDV), whereas the other is a polyclonal Ab raised in rabbits against rinderpest virus (RPV). Both Abs are known to show cross-reactivity with DMV. In this study we compared the labelling quality and the neuroanatomical distribution of staining with these two Abs by means of IHC analysis. To this end, serial sections of the target organs from ten free-ranging stranded cetaceans, previously diagnosed as being infected with DMV by PCR and/or serology, were subjected to IHC. The brain, lungs and lymph nodes of one animal were found to be positive with both Abs. From two other animals, the brain and the spleen, respectively, tested positive only with the polyclonal Ab. In the positive brain tissues, multifocal immunostaining was observed, with similar staining location and extent, with the two antibodies tested. Our results suggest that the polyclonal anti-RPV Ab might have a stronger binding activity to DMV than the anti-CDV MoAb. Nevertheless, the elaboration and use of specific anti-DMV Abs might be essential to guarantee conclusive results in diagnostic and pathogenetic investigations.

The Pathology of Cetacean Morbillivirus Infection and Comorbidities in Guiana Dolphins During an Unusual Mortality Event (Brazil, 2017-2018).

Cetacean morbillivirus (CeMV; Paramyxoviridae) is the most significant pathogen of cetaceans worldwide. The novel "multi-host" Guiana dolphin (Sotalia guianensis; GD)-CeMV strain is reported in South American waters and infects Guiana dolphins and southern right whales (Eubalaena australis). This study aimed to describe the pathologic findings, GD-CeMV viral antigen distribution and detection by RT-PCR (reverse transcriptase polymerase chain reaction), and infectious comorbidities in 29 Guiana dolphins that succumbed during an unusual mass-mortality event in Rio de Janeiro state, Brazil, between November 2017 and March 2018. The main gross findings were lack of ingesta, pulmonary edema, ascites, icterus, hepatic lipidosis, multicentric lymphadenomegaly, as well as pneumonia, polyserositis, and multiorgan vasculitis caused by Halocercus brasiliensis. Microscopically, the primary lesions were bronchointerstitial pneumonia and multicentric lymphoid depletion. The severity and extent of the lesions paralleled the distribution and intensity of morbilliviral antigen. For the first time in cetaceans, morbilliviral antigen was detected in salivary gland, optic nerve, heart, diaphragm, parietal and visceral epithelium of glomeruli, vulva, and thyroid gland. Viral antigen within circulating leukocytes suggested this as a mechanism of dissemination within the host. Comorbidities included disseminated toxoplasmosis, mycosis, ciliated protozoosis, and bacterial disease including brucellosis. These results provide strong evidence for GD-CeMV as the main cause of this unusual mass-mortality event.

Neurobrucellosis in a common bottlenose dolphin (Tursiops truncatus) stranded in the Canary Islands.

BACKGROUND: Brucella spp. isolation is increasingly reported in cetaceans, although associated pathologies, including lesions of the musculoskeletal and nervous systems, are less frequently described. Concerning the nervous system, Brucella sp. infection causing meningitis, meningoencephalitis or meningoencephalomyelitis have been extensively reported in striped dolphins (Stenella coeruleoalba), and less frequently in other cetacean species. CASE PRESENTATION: A juvenile female common bottlenose dolphin (Tursiops truncatus) was found stranded alive in Lanzarote (Canary Islands, Spain) in 2005, but died shortly after. On physical examination, the dolphin showed a moderate body condition and was classified as code 2 (fresh dead) at the time of necropsy. The main gross findings were severe multiorgan parasitism, thickened and congested leptomeninges, and (sero)fibrino-suppurative and proliferative arthritis of the shoulder joint. Histopathological examination revealed the distinct features of a sub-acute systemic disease associated with Cetacean Morbillivirus (CeMV) infection. However, brain lesions diverged from those reported in systemic CeMV infection. This led to suspect that there was a coinfecting pathogen, based on the characteristics of the inflammatory response and the lesion distribution pattern in the central nervous system. Brucella sp. was detected in the brain tissue by PCR and Brucella antigen was demonstrated by immunohistochemistry in the brain and shoulder joint lesions. CONCLUSIONS: The zoonotic potential of marine mammal strains of Brucella has been demonstrated both in natural and laboratory conditions. In this study, PCR detected Brucella sp. in the brain of a common bottlenose dolphin stranded in the Canary Islands; the dolphin was also co-infected with CeMV. This is the first detection of Brucella sp. infection in a stranded cetacean in this archipelago. Therefore, we stress the importance of taking adequate measures during the handling of these species to prevent the transmissions of the infection to humans.

Genetic heterogeneity of dolphin morbilliviruses detected in the Spanish Mediterranean in inter-epizootic period.

BACKGROUND: In the last 20 years, Cetacean Morbillivirus (CeMV) has been responsible for many die-offs in marine mammals worldwide, as clearly exemplified by the three dolphin morbillivirus (DMV) epizootics of 1990-1992, 2006-2008 and 2011 that affected Mediterranean striped dolphins (Stenella coeruleoalba). Systemic infection caused by DMV in the Mediterranean has been reported only during these outbreaks. RESULTS: We report the infection of five striped dolphins (Stenella coeruleoalba) stranded on the Spanish Mediterranean coast of Valencia after the last DMV outbreak that ended in 2011. Animal 1 stranded in late 2011 and Animal 2 in 2012. Systemic infection affecting all tissues was found based on histopathology and positive immunohistochemical and polymerase chain reaction positive results. Animal 3 stranded in 2014; molecular and immunohistochemical detection was positive only in the central nervous system. Animals 4 and 5 stranded in 2015, and DMV antigen was found in several tissues. Partial sequences of the DMV phosphoprotein (P), nucleoprotein (N), and hemagglutinin (H) genes were identical for Animals 2, 3, 4, and 5, and were remarkably different from those in Animal 1. The P sequence from Animal 1 was identical to that of the DMV strain that caused the epizootic of 2011 in the Spanish Mediterranean. The corresponding sequence from Animals 2-5 was identical to that from a striped dolphin stranded in 2011 on the Canary Islands and to six dolphins stranded in northeastern Atlantic of the Iberian Peninsula. CONCLUSIONS: These results suggest the existence of an endemic infection cycle among striped dolphins in the Mediterranean that may lead to occasional systemic disease presentations outside epizootic periods. This cycle involves multiple pathogenic viral strains, one of which may have originated in the Atlantic Ocean.

Prostatic Lesions in Odontocete Cetaceans.

The prostate is the only accessory male genital gland described in cetaceans. Although few studies describe the gross and histologic anatomy of the prostate in cetaceans, there is no information on pathological findings involving this organ. The prostate glands of 45 cetaceans, including 8 different odontocete species ( n = 44) and 1 mysticete, were evaluated. The main pathologic diagnoses were verminous prostatitis, septic prostatitis, viral prostatitis, benign prostatic hyperplasia, and prostatitis of unknown etiology. Verminous prostatitis ( n = 12) was caused by nematodes of the genus Crassicauda, and different presentations were observed. Septic prostatitis, identified in 2 cases, both involved nematode infestation and Clostridium spp coinfection. One case of viral prostatitis was identified and was associated with morbillivirus infection. In prostatitis of unknown cause ( n = 7), varying degrees of prostatic lesions, mostly chronic inflammation, were identified. Impacts at individual levels (eg, localized disease, loss of reproductive capacity) and population levels (eg, decreased reproductive success) are plausible. Our results indicate a high occurrence of prostatic lesions in free-ranging odontocetes. For this reason, the prostate should be routinely inspected and sampled during necropsy of odontocete cetaceans.

Pyogranulomatous obliterative laryngotracheitis by Rhizopus arrhizus (syn. R. oryzae) in a free-ranging Atlantic spotted dolphin Stenella frontalis.

We report the gross and microscopic findings and molecular identification of fungal hyphate infection in a juvenile female Atlantic spotted dolphin Stenella frontalis found dead off Arguineguin, Gran Canaria (Canary Islands, Spain). On necropsy examination, the animal had a large cranial intrathoracic mass and multiple variably-sized nodules throughout the larynx and trachea that obliterated the lumen. Microscopically, the masses were composed of abundant pyogranulomatous inflammation with numerous fungal hyphae. These were pauciseptate (coenocytic) and had non-parallel walls, non-dichotomous irregular to right angle branching, and bulbous dilations. PCR analysis from these inflammatory foci yielded Rhizopus arrhizus (syn. R. oryzae). This fungal pathogen is often ascribed to opportunistic infections in immunosuppressed humans and animals. In the present case, a potential cause for immunosuppression was not identified; PCR analysis for cetacean morbillivirus was negative. Herein, we report the first confirmed case of R. arrhizus infection in a free-living Atlantic cetacean. These findings add to the body of knowledge on fungal disease in cetaceans in general and, in particular, in odontocetes, where respiratory involvement is common.

Morbillivirus infection in Risso's dolphin Grampus griseus: a phylogenetic and pathological study of cases from the Canary Islands.

The earliest evidence of cetacean morbillivirus (CeMV) infection dates from 1982, when the dolphin morbillivirus strain (DMV) was identified in bottlenose dolphins Tursiops truncatus stranded in the mid-Atlantic region. Since then, CeMV has been detected globally in at least 26 species of mysticetes and odontocetes, causing widespread mortality and a wide range of pathological effects. In the Canary Islands, DMV and pilot whale morbillivirus have been detected in cetacean species, including short-finned pilot whales Globicephala macrorhynchus and bottlenose dolphins. Risso's dolphins Grampus griseus have been reported year-round in waters of the Canary Islands and are considered a resident species. No information is currently available on CeMV prevalence in this species in this ocean region. We searched for evidence of CeMV infection in 12 Risso's dolphins stranded in the Canary Islands from 2003 to 2015 by means of histopathology, PCR and immunohistochemistry. PCR revealed 2 CeMV-positive animals (16.6%). Phylogenetic analysis showed that the strains from the 2 positive specimens were phylogenetically quite distant, proving that more than 1 strain infects the Risso's dolphin population in this region. We also determined that the strain detected in one of the specimens mainly circulated in the northeastern Atlantic Ocean from 2007 to 2013.

Pulmonary carcinoma with metastasis in a long-finned pilot whale (Globicephala melas).

BACKGROUND: Lung cancer is the most commonly diagnosed neoplasm in humans, however this does not apply to other animal species. Living in an aquatic environment the respiratory system of cetaceans had to undergo unique adaptations in order to them to survive and cope with totally different respiratory pathogens and potentially carcinogens from those affecting humans. CASE PRESENTATION: This article discusses not only macroscopical, histopathological and immunohistochemical features of a pulmonary carcinoma with disseminated metastases in a long-finned pilot whale (Globicephala melas), as well as the immunohistochemical analysis performed on various tissues of cetaceans belonging to the genus Globicephala. On the necropsy examination of the carcass, multiple pulmonary nodules and generalised thoracic lymphadenomegaly were noted. Histologically, a malignant epithelial neoplasia was identified in the lung, thoracic lymph nodes, and adrenal gland. Immunohistochemical analysis revealed a pulmonary carcinoma. Vasculogenic mimicry and epithelial-to-mesenchymal transition phenotype, as suggested by cytomorphological and immunohistochemical characteristics, were observed. CONCLUSIONS: A diagnosis of metastatic pulmonary carcinoma was determined, which to the author's knowledge, appears to be not previously recorded in long-finned pilot whale species. This is also the first report of vasculogenic mimicry and epithelial-to-mesenchymal transition event in a spontaneous cancer from a cetacean species.

Fluorescent microbead-based immunoassay for anti-Erysipelothrix rhusiopathiae antibody detection in cetaceans.

A fluorescent microbead-based immunoassay (FMIA) for detection of anti-Erysipelothrix rhusiopathiae antibodies in pigs was adapted for use in cetaceans. The FMIA was validated and adjusted using serum samples from 10 vaccinated captive bottlenose dolphins Tursiops truncatus collected between 1 and 13 mo after immunization. The technique was then used to analyze specimens from 15 free-ranging cetaceans stranded alive on the Valencian Mediterranean coast between 2006 and 2014: 11 striped dolphins Stenella coeruleoalba, 3 Risso's dolphins Grampus griseus and 1 bottlenose dolphin Tursiops truncatus. One of these wild animals was confirmed to have died from E. rhusiopathiae septicemia, but no anti-E. rhusiopathiae antibodies were detected in its serum, pericardial fluid or milk samples. Another free-ranging individual, which lacked any signs or lesions that might be indicative of E. rhusiopathiae infection, showed high fluorescence intensity similar to that measured in captive dolphins at 6-13 mo after vaccination. These results suggest that this animal underwent an E. rhusiopathiae infection several months before stranding. The findings in the present study suggest that FMIA can be useful for detecting anti-E. rhusiopathiae antibodies in cetaceans, and its application to free-ranging animals is particularly interesting because of the great value of these specimens. Furthermore, the FMIA can be multiplexed to allow the determination of up to 100 analytes per sample in a single well, thereby reducing the cost, time and sample volume needed.

Fatal systemic morbillivirus infection in bottlenose dolphin, canary islands, Spain.

A systemic morbillivirus infection was diagnosed postmortem in a juvenile bottlenose dolphin stranded in the eastern North Atlantic Ocean in 2005. Sequence analysis of a conserved fragment of the morbillivirus phosphoprotein gene indicated that the virus is closely related to dolphin morbillivirus recently reported in striped dolphins in the Mediterranean Sea.

Retrospective study of etiologic agents associated with nonsuppurative meningoencephalitis in stranded cetaceans in the canary islands.

Nineteen natural cases of etiologically undetermined encephalitides in free-ranging cetaceans were studied retrospectively. Histological examination of the brains revealed variable degrees of nonsuppurative encephalitis or meningoencephalitis, characterized predominantly by perivascular lymphohistiocytic infiltrates. A PCR assay was used on brain and other available tissues to detect the presence of morbillivirus, herpesvirus, West Nile virus, Toxoplasma gondii, and Brucella spp. In addition, immunohistochemical (IHC) staining was performed on selected tissues to determine the presence of morbilliviral antigens. Six animals (5 striped dolphins and 1 common dolphin) showed IHC and/or molecular evidence of morbilliviral antigens and/or genomes, mainly in brain tissue. Conventional nested PCR detected herpesviral DNA in brain tissue samples from two striped dolphins. There was no evidence of West Nile virus, T. gondii, or Brucella spp. in any of the brain tissue samples examined. The information presented here increases the number of confirmed morbillivirus-positive cases within the Canarian archipelago from two previously reported cases to eight. Furthermore, a new nested-PCR method for the detection of morbillivirus is described here. Regarding herpesvirus, the phylogenetic analysis performed in the current study provides valuable information about a possible pathogenic branch of cetacean alphaherpesviruses that might be responsible for some fatal cases worldwide.

Case Report: Ambergris coprolite and septicemia in a male sperm whale stranded in La Palma (Canary Islands).

On the 21st of May 2023, a dead adult male sperm whale (Physeter macrocephalus) of 13 m in length and estimated weight of around 18,000 kg was reportedly stranded at Playa Los Nogales, La Palma, Canary Islands, Spain. A necropsy was performed 48hpm. A 50 cm diameter and 9.5 kg coprolite was found obstructing the caudal colon-rectal lumen. Necro-hemorrhagic lesions were found in heart muscles and three different bacteria of intestinal origin were isolated and identified (Edwarsiella tarda, Hathewaya limosa and Clostridium perfringens). It is reported a lethal septicemia of intestinal origin associated with ambergris coprolite as cause of death in this sperm whale.

Unusual striped dolphin mass mortality episode related to cetacean morbillivirus in the Spanish Mediterranean sea.

BACKGROUND: In the last 20 years, Cetacean Morbillivirus (CeMV) has been responsible for many die-offs in marine mammals worldwide, as clearly exemplified by the two dolphin morbillivirus (DMV) epizootics of 1990-1992 and 2006-2008, which affected Mediterranean striped dolphins (Stenella coeruleoalba). Between March and April 2011, the number of strandings on the Valencian Community coast (E Spain) increased. CASE PRESENTATION: Necropsy and sample collection were performed in all stranded animals, with good state of conservation. Subsequently, histopathology, immunohistochemistry, conventional reverse transcription polymerase chain reaction (RT-PCR) and Universal Probe Library (UPL) RT-PCR assays were performed to identify Morbillivirus. Gross and microscopic findings compatible with CeMV were found in the majority of analyzed animals. Immunopositivity in the brain and UPL RT-PCR positivity in seven of the nine analyzed animals in at least two tissues confirmed CeMV systemic infection. Phylogenetic analysis, based on sequencing part of the phosphoprotein gene, showed that this isolate is a closely related dolphin morbillivirus (DMV) to that responsible for the 2006-2008 epizootics. CONCLUSION: The combination of gross and histopathologic findings compatible with DMV with immunopositivity and molecular detection of DMV suggests that this DMV strain could cause this die-off event.

Pathology and causes of death of stranded cetaceans in the Canary Islands (1999-2005).

Between 1999 and 2005, 233 stranded cetaceans (comprising 19 species) were reported in the waters of the Canary Islands. Of these, 138/233 (59.2%) were subjected to a complete or partial standardized necropsy, including 4 Balaenopteridae, 9 Physeteridae, 8 Kogiidae, 27 Ziphiidae and 90 Delphinidae. Of these, 46/138 (33.3%) cetaceans were diagnosed with anthropogenic pathological categories (i.e. the cause of death was anthropogenic). These included fishing interaction (bycatch) (19 individuals), 'atypical' mass stranding events linked to naval exercises (13), ship collisions (8) and other anthropogenic-related pathology (6). 'Natural' (i.e. non-anthropogenic) causes of death accounted for another 82/138 (59.4%) cases, including infectious and non-infectious diseases (63), neonatal pathology (8), intra- and interspecific interactions (6) and mass strandings (5). The cause(s) of death could not be determined in 10/138 (7.3%) necropsied animals. The most common causes of death were ship collisions in 6/9 (66.6%) Physeteridae, 'atypical' mass stranding linked to naval exercises in 13/27 (48.1%) Ziphiidae, and 'natural' infectious and non-infectious diseases in 55/90 (61.1%) Delphinidae. Interaction with fishing activities was established as cause of death in 15/90 (16.7%) Delphinidae. These data show that a range of anthropogenic and natural single and mass mortality events occur in multiple cetacean species stranded in the Canary Islands.

Infection due to Buteo buteo herpesvirus in a common buzzard (Buteo buteo insularum).

This study aimed to document the pathological findings observed in a common buzzard (Buteo buteo insularum) from Gran Canaria (Canary Islands, Atlantic Ocean), naturally infected with Buteo buteo herpesvirus (HV). Local authorities found the common buzzard alive, but it died after 10 days of specialized veterinary care. Postmortem investigation, including complete gross and histologic examination, immunohistochemistry, microbiology, and PCR, was performed. The animal presented necrotizing heterophilic and histiocytic bilateral conjunctivitis, stomatitis, pharyngitis, rhinitis, and sinusitis with secondary bacterial and fungal infections. Frequent eosinophilic intranuclear inclusion bodies were observed in the oral mucosa and esophagus epithelium. HV proteins and DNA were detected in tissues from this animal. The sequences obtained from the PCR product were identical to the reported sequences of Buteo buteo HV.

Cerebral nasitremiasis in a Blainville's beaked whale (Mesoplodon densirostris) stranded in the Canary Islands.

Ten species within the genus Nasitrema (subfamily Nasitrematinae, family Brachycladiidae) have been reported infecting a wide variety of odontocetes worldwide, although there is still a lack of information about their presence in beaked whales (BWs). Nasitrema spp. are commonly described inhabiting the pterygoid sinus, the tympanic cavities, and the middle and inner ear; although aberrant migrations through the brain have been also reported. This trematode may cause different type of lesions, ranging from mild to severe saculitis, neuritis, otitis, and/or meningoencephalitis that may impede cetaceans to survive in the wild, resulting in incoordination, loss of equilibrium, and echolocation dysfunction ending in a stranding event. The presence of Nasitrema sp. was found in an adult female Blainville beaked whale stranded death in Fuerteventura, Canary Islands, on November 2016. The most relevant gross finding was a severe chronic-active multifocal pyogranulomatous and necrotizing encephalitis. Histologically, multiple areas of necrosis, pyogranulomatous and eosinophilic inflammation, haemorrhages and occasional cholesterol crystals were found associated with parasitic structures compatible with an adult trematode and its eggs. Molecular analysis, based on a quantitative polymerase chain reaction (qPCR) of the brain tissue sample detected 99% homology with a partial sequence of the NADH dehydrogenase subunit 3 (ND3) gene of Nasitrema delphini. In addition, liver, kidney, prescapular lymph node and brain samples were positive to herpesvirus (conventional nested PCR). Evidence of the presence of this parasite was not found in any of the 54 beaked whales (n = 54) stranded on the Canary Islands between 1999 and 2017, specifically 35 Cuvier's BWs and 19 specimens belonging to the Mesoplodon genus. To our knowledge, the current study represents the first description of a nasitremiasis in a member of the Ziphiidae family.

Viral skin diseases in odontocete cetaceans: gross, histopathological, and molecular characterization of selected pathogens.

Fifty-five skin lesions from 31 stranded cetaceans along the Canary coasts (2011-2021) were submitted to macroscopic, histological, and molecular analyses to confirm infection by cetacean poxvirus, herpesvirus and cetacean morbillivirus. They were macroscopically categorized into eight categories with respective subcategories according to their color, shape, size, and consistency. Cetacean poxvirus was detected in 54.54% of the skin lesions through real-time and conventional PCRs based on the DNA polymerase gene. Additionally, herpesvirus and morbillivirus were currently detected from 43.63 and 1.82% of the cutaneous lesions, respectively. Coinfection of poxvirus and herpesvirus was detected in nine of them (16.36%), which makes the present study the first to report coinfection by both pathogens in skin lesions in cetaceans. A plausible approach to histopathological characterization of poxvirus-and herpesvirus-positive skin lesions was established. Hyperkeratosis, acanthosis, ballooning degeneration, and intracytoplasmic inclusion bodies in vacuolized keratinocytes through the stratum spinosum were common findings in poxvirus skin lesions. Alphaherpesvirus was associated with a prominent acanthotic epidermis, moderate necrosis, multifocal dyskeratosis, and irregular keratinocytes with both cellular and nuclei pleomorphism. The common histopathological findings of both pathogens were observed in coinfection lesions. However, those associated with herpesvirus were considerably more remarkable. Relationships between molecular and microscopic findings were observed for the lesions that showed tattoo-like and tortuous patterns. Further multidisciplinary diagnostic studies of infected skin lesions are needed to understand the epidemiology of these emerging infectious diseases.

Cetacean Intracytoplasmic Eosinophilic Globules: A Cytomorphological, Histological, Histochemical, Immunohistochemical, and Proteomic Characterization.

The nature, etiopathogenesis, and clinicopathologic relevance of the prevalent intracytoplasmic eosinophilic globules (IEGs) within hepatocytes of cetaceans are unknown. This study aims to evaluate the presence and characterize the IEGs in the hepatocytes of cetaceans using histochemical and immunohistochemical electron microscopy, Western blot, lectin histochemistry, and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry techniques. A total of 95/115 (83%) animals (16 species) exhibited histologically evident intracytoplasmic round to oval, single to multiple, hyaline eosinophilic globules within the hepatocytes. These globules were largely PAS-positive, diastase resistant, and were immunopositive for fibrinogen (FB, 97%), albumin (Alb, 85%), and α1-antitrypsine (A1AT, 53%). The IEG positivity for FB and A1AT were correlated with live-stranding, hepatic congestion and a good nutritional status. The cetaceans lacking IEGs were consistently dead stranded and had poor body conditions. The IEGs in 36 bycaught cetaceans were, all except one, FB-positive and A1AT-negative. The IEGs exhibited morphologic and compositional variations at the ultrastructural level, suggesting various stages of development and/or etiopathogenesis(es). The glycocalyx analysis suggested an FB- and A1AT-glycosylation pattern variability between cetaceans and other animals. The proteomic analyses confirmed an association between the IEGs and acute phase proteins, suggesting a relationship between acute stress (i.e., bycatch), disease, and cellular protective mechanisms, allowing pathologists to correlate this morphological change using the acute hepatocytic cell response under certain stress conditions.