Morbillivirus Antigen Research Articles

Neuropathological Characterization of Dolphin Morbillivirus Infection in Cetaceans Stranded in Italy.

Simple SummaryThere is abundant literature reporting demyelination in dogs and pinnipeds affected by morbillivirus infection, but myelinopathy is poorly investigated in stranded cetaceans affected with the virus. Also, the neuropathogenesis of cetacean morbillivirus infection has not been fully clarified, leaving questions on cell tropism unanswered. A novel dolphin morbillivirus lineage of Atlantic origin circulating in Italian waters replaced the previous Mediterranean strain in late 2015; however, differences in virulence and pathogenesis between the two strains have not yet been documented. The aims of the present study were to: describe histopathological changes and immunohistochemical findings in the central nervous system of 31 cetaceans which tested positive on molecular investigations for the two dolphin morbillivirus strains; characterize by double indirect immunofluorescence staining the areas of myelin damage. The most frequently observed morbillivirus-associated lesions were astro-microgliosis, neuronal necrosis, spongiosis, malacia, and non-suppurative meningoencephalitis. Demyelination was detected by means of a specific myelin biomarker. Inside and around the demyelinated areas there were morbillivirus antigen-bearing cells of mainly neuronal and microglial origin, associated with marked astro and microglia reactivity. Molecular and immunohistochemical analysis suggested a higher neurotropic affinity of the novel circulating strain.Cetacean morbillivirus (CeMV) is responsible for epidemic and endemic fatalities in free-ranging cetaceans. Neuro-inflammation sustained by CeMV is a leading cause of death in stranded cetaceans. A novel dolphin morbillivirus (DMV) strain of Atlantic origin circulating in Italian waters since early 2016 has caused acute/subacute lesions associated with positive immunolabelling of the virus. To date, myelin damage has not been fully documented and investigated in cetaceans. This study describes neuropathological findings in the brain tissue of 31 cetaceans found stranded along the Italian coastline and positive for DMV infection on molecular testing. Cell changes in the areas of myelinopathy were revealed by double indirect immunofluorescence. The most frequent DMV-associated lesions were astro-microgliosis, neuronal necrosis, spongiosis, malacia, and non-suppurative meningoencephalitis. Myelin reduction and areas of demyelination were revealed by means of a specific myelin biomarker. Morbilliviral antigen immunolabelling was mainly observed in neurons and microglial cells, in association with a marked activation of microglia and astrocytes. These findings extend our knowledge of DMV-associated brain lesions and shed light on their pathogenesis.

Novel Presentation of DMV-Associated Encephalitis in a Long-Finned Pilot Whale (Globicephala melas)

Cetacean morbillivirus (CeMV) is an important global cause of morbidity and mortality in cetacean populations, with four pathological presentations including non-suppurative encephalitis. We describe an unusual case of dolphin morbillivirus (DMV)-associated non-suppurative encephalitis in a long-finned pilot whale (Globicephala melas), in which the lesions were orientated on the periventricular white matter and comprised prominent multifocal syncytia formation in the absence of systemic lesions. DMV RNA was detected in brain tissue by qRT-PCR and immunohistochemistry for morbillivirus antigen yielded intense labelling of syncytia in periventricular sites, with sparse involvement of the deeper neuroparenchyma. The pattern of lesions raises the possibility of viral dissemination through the cerebrospinal fluid, as described for canine distemper virus, suggesting that similar pathogenic mechanisms may be implicated in lesion development. Further investigation is required to establish the pathogenesis of CeMV encephalitis and the behaviour of the virus within the central nervous system of cetaceans.

Novel Dermatitis and Relative Viral Nucleic Acid Tissue Loads in a Fin Whale (Balaenoptera physalus) with Systemic Cetacean Morbillivirus Infection

Cetacean morbilliviruses (CeMVs) are significant causes of mortality in many cetacean species in epizootics and smaller outbreaks. Despite the prominence of skin lesions in seals and terrestrial animals, including humans, affected by other morbilliviruses, they have not been reported in CeMV-infected cetaceans. Here we report CeMV-associated skin lesions in a fin whale (Balaenoptera physalus) with subacute, systemic CeMV infection that live-stranded in Scotland, UK. Grossly, the skin was sloughing in large sheets, presumed due to autolysis, but histological examination showed syncytia, below the dermoepidermal junction, that were strongly immunopositive for morbillivirus antigen, as were syncytia in other organs. By polymerase chain reaction (PCR), the relative load of CeMV-specific RNA was largest in the liver and urinary bladder, even in formalin-fixed, paraffin-wax embedded samples. Levels were low in skin and only detectable in frozen samples. Genetic comparison of the CeMV revealed close alignment with isolates from fin whales from the North Atlantic Ocean and Mediterranean Sea, but that it was distinct from the porpoise CeMV clade. These findings show skin samples can be used to diagnose CeMV infection in cetaceans, highlighting the potential of ante-mortem sampling for monitoring disease in current populations and assessment of changes in host and pathogen genetics.

Cetacean Morbillivirus Infection in a Killer Whale (Orcinus orca) from Brazil

We provide pathological, immunohistochemical and molecular evidence of cetacean morbillivirus (CeMV) infection in a live-stranded adult female killer whale (Orcinus orca), which stranded alive in Espírito Santo State, Brazil, in 2014. Although attempts were made to release the animal, it stranded again and died. The main pathological findings were severe pulmonary oedema, pleural petechiation, multifocal, lymphoplasmacytic meningoencephalitis and leptomeningomyelitis with perivascular cuffing and gliosis, chronic lymphocytic bronchointerstitial pneumonia and multicentric lymph node and splenic lymphoid depletion. Other pathological findings were associated with the 'live-stranding stress response'. Immunohistochemical analysis revealed multifocal morbilliviral antigen in neurons and astrocytes, and in pneumocytes, histiocytes and leukocytes in the lung. CeMV was detected by a novel reverse transcriptase polymerase chain reaction method in the brain and kidney. Phylogenetic analysis of part of the morbillivirus phosphoprotein gene indicates that the virus is similar to the Guiana dolphin (Sotalia guianensis) morbillivirus strain, known to affect cetaceans along the coast of Brazil. To the authors' knowledge, this is the first report of morbillivirus disease in killer whales.

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.

Pathological Findings in White-Beaked Dolphins (Lagenorhynchus albirostris) and Atlantic White-Sided Dolphins (Lagenorhynchus acutus) From the South-Eastern North Sea.

In the North Sea, white-beaked dolphins (Lagenorhynchus albirostris) occur regularly and are the second most common cetacean in the area, while their close relative, the Atlantic white-sided dolphin (Lagenorhynchus acutus), prefers the deeper waters of the northern North Sea and adjacent Atlantic Ocean. Though strandings of both species have occurred regularly in the past three decades, they have decreased in the southern North Sea during the last years. Studies describing necropsy findings in stranded Lagenorhynchus spp. are, to date, still scarce, while information gained through post-mortem examinations may reveal valuable information about underlying causes of this decline, including age structure and the reproduction status. Therefore, we retrospectively assessed and compared the necropsy results from fresh Lagenorhynchus spp. stranded along the southeastern North Sea between 1990 and 2019. A full necropsy was performed on 24 white-beaked dolphins and three Atlantic white-sided dolphins from the German and Dutch coast. Samples of selected organs were taken for histopathological, bacteriological, mycological, parasitological and virological examinations. The most common post-mortem findings were emaciation, gastritis and pneumonia. Gastritis and ulceration of the stomach was often associated with an anisakid nematode infection. Pneumonia was most likely caused by bacterial infections. Encephalitis was observed in three animals and morbillivirus antigen was detected immunohistochemically in one case. Although the animal also showed pneumonic lesions, virus antigen was only found in the brain. Parasitic infections mainly affected the gastro-intestinal tract. Lungworm infections were only detected in two cases and no associations with pathological alterations were observed. Stenurus spp. were identified in two of three cases of parasitic infections of the ears. Twelve of the 26 white-beaked dolphins stranded in Germany were found between 1993 and 1994, but there was no evidence of epizootic disease events or mass strandings during the monitored period.

Pathologic findings and causes of death in southern right whales Eubalaena australis, Brazil.

Southern right whales Eubalaena australis (SRWs) migrate to southern Brazil for breeding and calving from June through November. Overall, there is scarce knowledge on health status and pathologic conditions in SRWs. We report the pathologic and molecular investigation results of 8 SRWs that were necropsied between 2010 and 2017 within a breeding and calving ground in Santa Catarina state, Brazil. The animals were of various ages (7 newborns/calves, 1 adult) and sex (3 females, 5 males). Five whales stranded dead; 3 stranded alive and died shortly after (n = 2) or were euthanized (n = 1). The causes of stranding and/or death were neonatal respiratory distress syndrome with meconium aspiration (n = 3) with concomitant congenital hepatopathy in one of them; trauma of unknown origin (n = 3), infectious renal and lung disease with presumed sepsis (n = 1), and euthanasia (n = 1). Three animals were PCR-positive for cetacean morbillivirus; one of them also had morbilliviral antigen in kidney via immunohistochemical analysis. These results, integrating novel findings and a published report, contribute to the pathology knowledge of this species.

Morbillivirus and Pilot Whale Deaths, Canary Islands, Spain, 2015

Morbillivirus and Pilot Whale Deaths, Canary Islands, Spain, 2015

Cetacean morbillivirus in Northern and Southern Hemispheres.

In the last 25 years, at least 10 dramatic morbilliviral epidemics have occurred among free-ranging pinniped and cetacean species and populations worldwide. The origin(s) of the new Morbillivirus genus members causing these mass mortality events, along with the reason(s) behind their “sudden” appearance among wild aquatic mammals, are still unknown (Di Guardo et al., 2005, 2011; Di Guardo, 2012). Interestingly enough, two recent papers have reported the existence of Cetacean morbillivirus (CeMV) strains infecting wild cetaceans (Tursiops aduncus and Sotalia guianensis, respectively) in the Southern Hemisphere (Groch et al., 2014; Stephens et al., 2014) and exhibiting marked genetic differences from those infecting cetaceans of the Northern Hemisphere (Van Bressem et al., 2009; Di Guardo et al., 2011; Di Guardo and Mazzariol, 2013). Albeit quantified only for the nucleoprotein (N) and phosphoprotein (P) genes, these differences are so consistent that one could question the inclusion of the concerned isolates into the CeMV clade. Furthermore, the agent recovered from the two T. aduncus individuals stranded along the Western Australia coastline was deemed to be, with exclusive reference to the N and P genes, the CeMV more closely related to Measles Virus (MeV) as well as to Peste des Petits Ruminants Virus (PPRV) and Rinderpest Virus (RPV), the archetype morbillivirus (Stephens et al., 2014). Intriguingly, no peculiar inflammatory lesions nor immunohistochemical (IHC) evidence of morbilliviral antigen were observed in the brain of these two animals (Stephens et al., 2014), differently from CeMV-infected cetaceans (Di Guardo and Mazzariol, 2013; Di Guardo et al., 2013) but also similarly to experimentally RPV-infected cattle (Wohlsein et al., 1993). In this respect, while CeMV sequences could be demonstrated in the brain and lung from approximately 40% of the striped dolphins (Stenella coeruleoalba) found stranded during the first mid of 2013 along the Tyrrhenian coast of Italy, IHC evidence of morbilliviral antigen was obtained from the brain of only one of these animals (Casalone et al., 2014). Nevertheless, the isolate(s) characterized from these dolphins, apart from being identical to each other, were also very similar to those responsible for the three previous epidemic outbreaks occurred throughout the last 25 years in the Western Mediterranean (Di Guardo and Mazzariol, 2013; Casalone et al., 2014). In conclusion, while more data are certainly needed on the genetic “make-up” of the newly discovered CeMV strains infecting cetaceans in the Southern Hemisphere (Groch et al., 2014; Stephens et al., 2014), it should be additionally emphasized that, despite the existence of single, highly valuable contributions in this field (Shimizu et al., 2013), a very limited body of knowledge is available on the host-related components as well as on the agent-related factors driving the complex Morbillivirus-cetacean interaction dynamics (Di Guardo, 2012).

Is Dolphin Morbillivirus Virulent for White-Beaked Dolphins (Lagenorhynchus albirostris)?

The virulence of morbilliviruses for toothed whales (odontocetes) appears to differ according to host species. In 4 species of odontocetes, morbilliviruses are highly virulent, causing large-scale epizootics with high mortality. In 8 other species of odontocetes, including white-beaked dolphins (Lagenorhynchus albirostris), morbilliviruses have been found as an incidental infection. In these species, the virulence of morbilliviruses is not clear. Therefore, the admission of 2 white-beaked dolphins with morbillivirus infection into a rehabilitation center provided a unique opportunity to investigate the virulence of morbillivirus in this species. By phylogenetic analysis, the morbilliviruses in both animals were identified as a dolphin morbillivirus (DMV) most closely related to that detected in a white-beaked dolphin in Germany in 2007. Both animals were examined clinically and pathologically. Case No. 1 had a chronic neural DMV infection, characterized by polioencephalitis in the cerebrum and morbillivirus antigen expression limited to neurons and glial cells. Surprisingly, no nervous signs were observed in this animal during the 6 months before death. Case No. 2 had a subacute systemic DMV infection, characterized by interstitial pneumonia, leucopenia, lymphoid depletion, and DMV antigen expression in mononuclear cells and syncytia in the lung and in mononuclear cells in multiple lymphoid organs. Cause of death was not attributed to DMV infection in either animal. DMV was not detected in 2 contemporaneously stranded white-beaked dolphins. Stranding rate did not increase in the region. These results suggest that DMV is not highly virulent for white-beaked dolphins.

Morbillivirus infection in cetaceans stranded along the Italian coastline: Pathological, immunohistochemical and biomolecular findings

Morbilliviruses are recognized as biological agents highly impacting the health and conservation status of free-ranging cetaceans worldwide, as clearly exemplified by the two Dolphin Morbillivirus (DMV) epidemics of 1990–1992 and 2006–2008 among Mediterranean striped dolphins (Stenella coeruleoalba). After these two epidemics, morbilliviral infection (MI) cases with peculiar neurobiological features were reported in striped dolphins stranded along the Spanish coastline. Affected cetaceans showed a subacute-to-chronic, non-suppurative encephalitis, with brain lesions strongly resembling those found in human “subacute sclerosing panencephalitis” and “old dog encephalitis”. Brain was the only tissue in which morbilliviral antigen and/or genome could be detected.Beside a case of morbilliviral encephalitis in a striped dolphin’s calf stranded in 2009, we observed 5 additional MI cases in 2 striped dolphins, 1 bottlenose dolphin (Tursiops truncatus) and 2 fin whales (Balaenoptera physalus), all stranded in 2011 along the Italian coastline. Noteworthy, 3 of these animals (2 striped dolphins and 1 bottlenose dolphin) showed immunohistochemical (IHC) and/or biomolecular (PCR) evidence of morbilliviral antigen and/or genome exclusively in their brain, with 1 striped dolphin and 1 bottlenose dolphin also exhibiting a non-suppurative encephalitis. Furthermore, simultaneous IHC and PCR evidence of a Toxoplasma gondii coinfection was obtained in 1 fin whale.The above results are consistent with those reported in striped dolphins after the two MI epidemics of 1990–92 and 2006–2008, with evidence of morbilliviral antigen and/or genome being found exclusively in the brain tissue from affected animals.

Morbillivirus-host interaction: lessons from aquatic mammals

GENERAL COMMENTARY article Front. Microbiol., 24 December 2012Sec. Virology Volume 3 - 2012 | https://doi.org/10.3389/fmicb.2012.00431

Fatal cetacean morbillivirus infection in an Australian offshore bottlenose dolphin (Tursiops truncatus)

A juvenile offshore bottlenose dolphin (Tursiops truncatus) was found stranded with neurological signs and unable to swim or float unassisted. It subsequently died, succumbing to a combination of severe pneumonia and encephalitis. Morbillivirus serum neutralisation test serology was positive (titre 1:16) for cetacean morbillivirus and negative for both phocine distemper virus and canine distemper virus. There was concurrent thymic and lymph node lymphoid depletion and necrosis, together with intranuclear and intracytoplasmic acidophilic viral inclusion bodies and multinucleate syncytia within multiple organs. Paramyxovirus capsids were identified in lung sections via electron microscopy and morbillivirus antigen was demonstrated within sections of lung, thymus and brain by immunohistochemistry. Reverse transcription-polymerase chain reaction for morbillivirus nucleoprotein (N) and phosphoprotein (P) genes were positive and phylogenetic gene product sequence analysis revealed 98% and 94% sequence identity to dolphin morbillivirus, respectively. To the authors' knowledge, this is the first report of a cetacean mortality due to morbillivirus infection occurring in the southern hemisphere. Morbillivirus infection should be included in the differential diagnosis of stranded live or dead cetaceans in Australian waters, particularly if animals display neurological signs.

Morbilliviral encephalitis in a striped dolphin Stenella coeruleoalba calf from Italy

A free-living, striped dolphin Stenella coeruleoalba calf was stranded on the Latium coast of Italy in November 2009. Significant neuropathological findings included non-suppurative meningoencephalitis, microgliosis, neuronal degeneration, astrocytosis, and occasional multinucleate syncytia. Immunohistochemistry (IHC) and reverse transcription polymerase chain reaction (RT-PCR) for Morbillivirus were positive exclusively from the brain, with morbilliviral antigen and nucleic acid being detected in neurons and, to a lesser extent, in astrocytes. A low neutralizing antibody titer (1:10) against Morbillivirus (Canine distemper virus, CDV) was also found in blood serum, with no simultaneous presence of serum antibodies to Brucella spp. or Toxoplasma gondii. Furthermore, no pathogenic bacteria were isolated from any tissue or biological sample. This is the second report of morbilliviral encephalitis in a striped dolphin stranded along the Italian coastline in a 16 yr period (1993 to 2009). The neurohistopathological, IHC, and biomolecular features of this case are of additional interest, as antigenic and genomic positivity were exclusively confined to the brain of this dolphin, which may have acquired morbilliviral infection either postnatally or transplacentally.

Systemic Herpesvirus and Morbillivirus Co-Infection in a Striped Dolphin (Stenella coeruleoalba)

During 2007 a dolphin morbillivirus epizootic affected the western Mediterranean and several striped dolphins (Stenella coeruleoalba) stranded on the Catalonian coasts. One of those animals had severe lymphoid depletion, necrosis and syncytial formation in lymph nodes and spleen, with large basophilic nuclear inclusions compatible with herpesvirus detected by immunohistochemical and ultrastructural examination. Non-suppurative encephalitis with associated morbillivirus antigen and morbillivirus antigen within alveolar macrophages were also observed. A pan-herpesvirus nested polymerase chain reaction amplified a sequence virtually identical to two cetacean herpesvirus sequences previously identified in systemic infections in an Atlantic Cuvier's beaked whale (Ziphius cavirostris) and in a Mediterranean striped dolphin. The herpesviral infection was probably secondary to the immunosuppression caused by the morbillivirus. To our knowledge, this is the first report of a cetacean co-infected by dolphin morbillivirus and herpesvirus with evidence of lesions attributable to both viruses.

Chronic Morbilliviral Encephalitis in a Young Striped Dolphin from Italy

AbstractA young male striped dolphin (Stenella coeruleoalba) found stranded in November 2009 was affected by a chronic non-suppurative encephalitis, with prominent mononuclear cell perivascular cuffs, neuronal degeneration, microgliosis, neuronophagia and occasional presence of multinucleate syncytia. Immunohistochemical and biomolecular investigations for Morbillivirus were positive exclusively from the brain, but not from the cerebellum nor from any other tissue, with morbilliviral antigen being detected in neurons and astrocytes. A low neutralizing antibody titer (1:10) against Morbillivirus (CDV) was also found, with no simultaneous presence of anti-Brucella antibodies.The main reason why the present case appears to be of interest is provided by its peculiar neuro-histopathologic, immunohistochemical and biomolecular features, with Morbillivirus antigenic and genomic positivity being exclusively confined to the brain of this dolphin, which may have developed a transplacentally-acquired infection.

Cerebral Toxoplasmosis in Striped Dolphins (Stenella coeruleoalba) Stranded Along the Ligurian Sea Coast of Italy

This article reports the results of necropsy, parasitologic, microbiologic, histopathologic, immunohistochemical, indirect immunofluorescence, biomolecular, and serologic investigations on 8 striped dolphins (Stenella coeruleoalba) found stranded from August to December 2007 on the Ligurian Sea coast of Italy. Severe, nonsuppurative meningoencephalitis was found in 4 animals, as characterized by prominent perivascular mononuclear cell cuffing and macrophage accumulations in neuropil. These lesions were associated with mild lymphocytic-plasmacytic infiltration of choroid plexuses in 1 dolphin. Toxoplasma gondii cysts and zoites, confirmed by immunohistochemical labeling, were scattered throughout the brain parenchyma of 2 of the 4 dolphins. No viral inclusions were seen in the brain of any animal. Other findings included severe bronchointerstitial pneumonia and pulmonary atelectasis, consolidation, and emphysema. Parasites were identified in a variety of organs, including lung (Halocerchus lagenorhynchi). Microbiologic and serologic examinations for Brucella spp were negative on all 8 dolphins. The 4 animals with meningoencephalitis had serum antibodies against T gondii (titers ranging from 1:80 to 1:320) but not against morbillivirus. In contrast, the other 4 dolphins were seropositive for morbillivirus (with titers ranging from 1:10 to 1:40) but seronegative for T gondii. No morbillivirus antigen or nucleic acid was detected in the tissues of any dolphin. It is concluded that the severe lung and brain lesions were the cause of death and that T gondii was the likely etiologic agent of the cerebral lesions. Morbillivirus infection was not considered to have contributed to death of these animals.

Morbillivirus infection in free-ranging Atlantic bottlenose dolphins ( Tursiops truncatus) from the Southeastern United States: Seroepidemiologic and pathologic evidence of subclinical infection

From 2003 to 2007, sera ( n = 234) from free-ranging Atlantic bottlenose dolphins ( Tursiops truncatus) inhabiting two southeast Atlantic estuarine regions, the Indian River Lagoon (IRL), FL and Charleston, SC (CHS) were tested for antibodies to cetacean morbilliviruses as part of a multidisciplinary study of individual and population health. Positive morbillivirus titers were found on initial capture in 12 of 122 (9.8%) IRL dolphins in the absence of an epizootic. All CHS dolphins were seronegative. Positive fluctuating morbillivirus titers and seroconversion were found in IRL dolphins. Seropositivity was detected in dolphins 8–13 years of age as well as in dolphins that were alive during the 1987–1988 epizootic. During the study period, pathologic and immunohistochemical findings from stranded IRL dolphins ( n = 14) did not demonstrate typical morbillivirus-associated lesions or the presence of morbillivirus antigen. The findings suggest that morbillivirus infections are occurring in the absence of widespread mortality in IRL dolphins.

Morbillivirus Infections in Aquatic Mammals: A Brief Overview

Since 1987, at least eight morbillivirus infection (MI) epidemics have caused mass mortality of several free-living pinniped and cetacean populations around the world. The responsible agents, all belonging to the genus Morbillivirus (family Paramyxoviridae), have been characterized as either "canine distemper virus" strains, infecting pinnipeds, or as three new morbilliviruses, namely "phocid (phocine) distemper virus" , "porpoise morbillivirus" and "dolphin morbillivirus" . The last two agents are currently gathered under the common denomination of "cetacean morbillivirus". At post-mortem examination, a commonly occurring macroscopic lesion is represented by more or less severe bilateral pneumonia, with consolidation, congestion and oedema of both lungs, which fail to collapse. Histologically, a non-suppurative broncho-interstitial pneumonia, characterized by type II pneumocyte hyperplasia and by formation of endobronchial, endobronchiolar and endoalveolar "Warthin-Finkeldey type" syncytia, as well as a multifocal, non-suppurative encephalitis, associated with a severe and generalized lymphoid tissue depletion, are common pathological findings. Furthermore, eosinophilic viral inclusions are often detected, at both the intracytoplasmic and intranuclear level, within bronchial and bronchiolar epithelial, pulmonary syncytial, neuronal and other cell types. These inclusions, along with lymphoid and other cellular elements, are often found to be immunohistochemically positive for morbillivirus antigen. Among the still debated, or even controversial issues regarding MI in sea mammals, the one related to the origin of their causative agents is of particular concern. Another intriguing issue regards the synergistic effects, if any, associated with chronic exposure to a number of environmental pollutants, such as organochlorines and heavy metals. In fact, it is also unknown whether and how these chemicals contribute towards modulating the pathogenic and pathogenetic activity primarily displayed by sea mammal morbilliviruses.

Immunohistological and serological investigations of morbillivirus infection in Black Sea harbour porpoises ( Phocoena phocoena)

In the present study the occurrence of morbillivirus infection in harbour porpoises ( Phocoena phocoena) from the Black Sea was investigated. Blood and tissue specimens of lung, brain and spleen from 73 stranded or by-caught harbour porpoises derived from the three Black Sea subregions such as Bulgaria, Georgia and Ukraine were collected between 1997 and 1999 and processed for histology, immunohistochemistry and serology. Age determination was performed according to dental growth layers and body length. The age of the investigated population ranged from neonates to a 12-year-old animal. Morbillivirus-specific neutralizing antibodies were detected in 53% of harbour porpoises. Generally, titres were very low and ranged from 20 to 270. There was no correlation between age, geographical origin and titre levels. The most common histological finding (97%) consisted of a mild to severe granulomatous bronchopneumonia due to lung worm infection. There were no changes indicative of a morbillivirus infection. Using immunohistology none of the animals were positive for morbillivirus antigen. However, the serological data are suggestive of a continuously circulating morbillivirus among harbour porpoises from the Black Sea indicating that harbour porpoises may serve as carriers for fatal diseases in susceptible cetacean species.