Manuel Arbelo

Gas-bubble lesions in stranded cetaceans

There are spatial and temporal links between some mass strandings of cetaceans — predominantly beaked whales — and the deployment of military sonar. Here we present evidence of acute and chronic tissue damage in stranded cetaceans that results from the formation in vivo of gas bubbles, challenging the view that these mammals do not suffer decompression sickness. The incidence of such cases during a naval sonar exercise indicates that acoustic factors could be important in the aetiology of bubble-related disease and may call for further environmental regulation of such activity.

Insights into the diet of beaked whales from the atypical mass stranding in the Canary Islands in September 2002

Stomach contents were analysed from three species of beaked whales which mass-stranded shortly after a naval exercise conducted in the Canary Islands in September 2002. Animals from such mass strandings often contain freshly ingested food in their stomachs and can provide a more reliable guide to feeding habits than other strandings. Food remains recovered from seven Cuvier’s beaked whales (Ziphius cavirostris) consisted mainly of oceanic cephalopods, the most numerous being Taonius pavo, Histioteuthis sp., Mastigoteuthis schmidti and Octopoteuthis sicula. Many of the cephalopod species found in the diet appear to undertake daily vertical migrations, being found in shallower waters during the night and moving to deeper waters during the day. Single specimens of Blainville’s beaked whale (Mesoplodon densirostris) and Gervais’ beaked whale (Mesoplodon europaeus) had eaten both fish and cephalopod prey. The most numerous prey remains belonged to gadid fish and viperfish (Chauliodus sp.) respectively. These results are consistent with the limited published data on diet in these species, with Mesoplodon species having a relatively higher proportion of fish in the diet whereas Ziphius specialises on cephalopods.

Retrospective Study of Etiologic Agents Associated with Nonsuppurative Meningoencephalitis in Stranded Cetaceans in the Canary Islands

ABSTRACT 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.

Decompression vs. Decomposition: Distribution, Amount, and Gas Composition of Bubbles in Stranded Marine Mammals

Gas embolic lesions linked to military sonar have been described in stranded cetaceans including beaked whales. These descriptions suggest that gas bubbles in marine mammal tissues may be more common than previously thought. In this study we have analyzed gas amount (by gas score) and gas composition within different decomposition codes using a standardized methodology. This broad study has allowed us to explore species-specific variability in bubble prevalence, amount, distribution, and composition, as well as masking of bubble content by putrefaction gases. Bubbles detected within the cardiovascular system and other tissues related to both pre- and port-mortem processes are a common finding on necropsy of stranded cetaceans. To minimize masking by putrefaction gases, necropsy, and gas sampling must be performed as soon as possible. Before 24 h post mortem is recommended but preferably within 12 h post mortem. At necropsy, amount of bubbles (gas score) in decomposition code 2 in stranded cetaceans was found to be more important than merely presence vs. absence of bubbles from a pathological point of view. Deep divers presented higher abundance of gas bubbles, mainly composed of 70% nitrogen and 30% CO2, suggesting a higher predisposition of these species to suffer from decompression-related gas embolism.

Phylogenetic analysis of a new Cetacean morbillivirus from a short-finned pilot whale stranded in the Canary Islands

Cetacean morbillivirus (CeMV) is considered the most pathogenic virus in cetaceans. Three strains have been already described: the dolphin morbillivirus (DMV), the porpoise morbillivirus (PMV) and the tentatively named pilot whale morbillivirus (PWMV). This study describes the molecular characterization of a strain of CeMV detected in the brain of a short-finned pilot whale that had stranded in the Eastern Atlantic Ocean around the Canary Islands and that showed lesions compatible with morbilliviral disease. Sequences for the nucleoprotein, phosphoprotein, fusion protein and haemagglutinin genes were obtained. The phylogenetic study showed high homology (97%) with the PWMV strain previously detected from a long-finned pilot whale stranded in the Western Atlantic Ocean. These results support the existing classification of CeMV into three principal genetic clusters.

Decompression sickness (‘the bends’) in sea turtles

Decompression sickness (DCS), as clinically diagnosed by reversal of symptoms with recompression, has never been reported in aquatic breath-hold diving vertebrates despite the occurrence of tissue gas tensions sufficient for bubble formation and injury in terrestrial animals. Similarly to diving mammals, sea turtles manage gas exchange and decompression through anatomical, physiological, and behavioral adaptations. In the former group, DCS-like lesions have been observed on necropsies following behavioral disturbance such as high-powered acoustic sources (e.g. active sonar) and in bycaught animals. In sea turtles, in spite of abundant literature on diving physiology and bycatch interference, this is the first report of DCS-like symptoms and lesions. We diagnosed a clinico-pathological condition consistent with DCS in 29 gas-embolized loggerhead sea turtles Caretta caretta from a sample of 67. Fifty-nine were recovered alive and 8 had recently died following bycatch in trawls and gillnets of local fisheries from the east coast of Spain. Gas embolization and distribution in vital organs were evaluated through conventional radiography, computed tomography, and ultrasound. Additionally, positive response following repressurization was clinically observed in 2 live affected turtles. Gas embolism was also observed postmortem in carcasses and tissues as described in cetaceans and human divers. Compositional gas analysis of intravascular bubbles was consistent with DCS. Definitive diagnosis of DCS in sea turtles opens a new era for research in sea turtle diving physiology, conservation, and bycatch impact mitigation, as well as for comparative studies in other air-breathing marine vertebrates and human divers.

Presence of herpesvirus in striped dolphins stranded during the cetacean morbillivirus epizootic along the Mediterranean Spanish coast in 2007

A screening for herpesvirus (HV) was carried out using a tissue bank obtained from the cetacean morbillivirus (CeMV) mortality episode that occurred along the Mediterranean Spanish coast in 2007. A total of 14 cetaceans, including six long-finned pilot whales and eight striped dolphins, were studied using histopathology and molecular analysis to detect HV and CeMV. In five of the eight dolphins (62.5%) infected with CeMV, eight novel HV sequences were also detected. No HV lesions were found in any of the coinfected dolphins, which may indicate that HV did not contribute to the mortality in the CeMV outbreak. This is the first report of HV infection in any cetacean from the Mediterranean Sea.

Rhabdomyolysis and Myoglobinuric Nephrosis (Capture Myopathy) in a Striped Dolphin

This report describes delayed myoglobinuric capture myopathy in a striped dolphin (Stenella coeruleoalba) found stranded alive on the coast of Fuerteventura (Canary Islands, Spain). The animal was transported to Gran Canaria where it died 48 hr after stranding. The main lesions consisted of acute rhabdomyolysis affecting both cardiac and skeletal muscles, and myoglobinuric nephrosis. Using immunohistochemistry, degenerate myofibers with depletion of myoglobin, and an intracytoplasmatic immunoreaction for fibrinogen were observed. Orange-red pigmented casts in renal tubular lumens were strongly immunolabeled for myoglobin. To our knowledge, this is the first pathologic description of capture myopathy with myoglobinuric nephrosis in stranded cetaceans. Stress, exertion, trauma, and crush injury caused during the stranding, restraint, and transportation were the main causes of rhabdomyolysis in this case.

Immunophenotypic characterization of hepatic inflammatory cell infiltrates in common dolphins (Delphinus delphis)

Of 14 common dolphins, 12 showed non-specific reactive hepatitis and three chronic parasitic cholangitis with lymphoid proliferation. Non-specific reactive hepatitis was shown immunohistochemically to be associated with small clusters of CD3(+) cells in portal areas and hepatic sinusoids. Polyclonal antibody against S100 protein reacted with a variable number of lymphocytes from portal areas and hepatic sinusoids, as well as with Kupffer cells and epithelial cells of the bile ducts. The majority of plasma cells observed in portal areas and hepatic sinusoids were IgG(+). In lymphonodular lesions of chronic parasitic cholangitis, the distribution of immunoreactive cells was similar to that found in the cortex of lymph nodes. The presence of stellate cells similar to follicular dendritic and interdigitating cells expressing S-100 protein and MHC class II antigen in lymphonodular lesions suggested that these were highly organized structures developed to enhance antigen presentation to B and T cells.

Cross-reactivity of human and bovine antibodies in striped dolphin paraffin wax-embedded tissues.

This study evaluates the cross-reactivity of seven anti-human and one anti-bovine antibodies in formalin-fixed, paraffin-embedded tissue samples of liver and mesenteric lymph nodes of 13 striped dolphins (Stenella coeruleoalba). Four antibodies (CD3, IgG, lysozyme and S100 protein) reacted with striped dolphin lymph nodes in a similar pattern to that observed in the species of origin. The anti-human MHC class II mAb reacted strongly with macrophages and dendritic-like cells of striped dolphins, whereas a small number of lymphocytes were labelled with this antibody. These antibodies were used to study the immunophenotype of the inflammatory infiltrated in non-specific chronic reactive hepatitis (eight cases) and chronic parasite cholangitis (two cases) and normal liver (three cases) of striped dolphins. Non-specific chronic reactive hepatitis was composed of inflammatory infiltration of CD3+ T lymphocytes and IgG+ plasma cells in portal spaces and hepatic sinusoids. Lymphonodular aggregates observed in chronic parasitic cholangitis showed a cellular distribution similar to that found in lymph node cortex, including the presence of S100+ and MHC class II+ dendritic-like cells in lymphoid follicles and interfollicular areas. This result suggests that those inflammatory infiltrates are highly organised to enhance antigen presentation to B and T cells.