New colonisers drive the increase of the emerging loggerhead turtle nesting in Western Mediterranean.

The loggerhead sea turtle (Caretta caretta) is sensitive to climate change and is responding by colonising the Western Mediterranean. To understand the rapid nesting increase in recent years in Spain, we sampled 45 hatchlings from 8 nests between 2016 and 2019. We sequenced a mtDNA D-loop region, genotyped 2291 SNPs using 2bRAD and collected data on clutch size, hatching success, and incubation duration. We confirmed that the colonisation has a Mediterranean and Atlantic mixed origin and we detected that these nests were laid by different females, except for two nests within the same season. Our results suggest that the recent increase in nesting is due to an increase in the number of colonising individuals rather than females born in the same area returning to breed. We hypothesize that this increase in the number of colonisers results from successful conservation efforts, feminisation of the populations of origin and earlier sexual maturation. However, the percentage of offspring females produced in Spain suggests that future returning individuals will aid to the settlement of the new population. These results allow defining the current status of this colonisation although future efforts are needed to detect remigrants to confirm the establishment of a resident population.

New insights into the pathogenesis and transmission of Brucella pinnipedialis: systemic infection in two bottlenose dolphins (Tursiops truncatus).

IMPORTANCE: Brucella spp. are zoonotic pathogens that can affect both terrestrial and marine mammals. Brucella ceti has been identified in various cetacean species, but only one sequence type (ST27) has been reported in humans. However, it is important to conduct surveillance studies to better understand the impact of marine Brucella species on marine mammals, a typically understudied host group. Here, we describe a systemic infection by two related strains of Brucella pinnipedialis (ST25) in a couple of live-stranded bottlenose dolphins, with more severe lesions in the younger animal. Furthermore, B. pinnipedialis was first detected in milk from a female cetacean that stranded with its offspring. Our study reveals novel insights into the epidemiology and pathological consequences of B. pinnipedialis infections in cetaceans, emphasizing the crucial importance of ongoing surveillance and accurate diagnosis to understand the impact of this pathogen on marine mammal populations.

Tension pneumothorax in small odontocetes.

Pneumothorax, the accumulation of air in the pleural cavity, occurs when air enters the pleural space by the pleuro-cutaneous, pleuro-pulmonary, or pleuro-oesophageal-mediastinal route. Tension pneumothorax is an infrequent and severe form of pneumothorax where a positive pressure in the pleural space is built up during at least part of the respiratory cycle, with compression of both lungs and mediastinal vessels, and, if unilateral, with midline deviation towards the unaffected hemithorax. We describe 9 cases of tension pneumothorax in 3 species of small cetaceans (striped dolphin Stenella coeruleoalba, common dolphin Delphinus delphis, and common bottlenose dolphin Tursiops truncatus) from the western Mediterranean coast of Spain, and one case from a dolphinarium. Computed tomography (CT) imaging performed in 2 carcasses before necropsy showed lung compression, midline deviation, and pressure on the diaphragm, which was caudally displaced. Tension pneumothorax was recognized at necropsy by the presence of pressurized air in one of the hemithoraces. Seven of the pneumothorax cases were spontaneous (2 primary and 5 secondary to previous lung pathology). In the other 2 dolphins, the pneumothorax was traumatic, due to oesophageal-pleural perforation or rib fractures. We hypothesize that pneumothorax in dolphins is predominantly tensional because of their specific anatomical and physiological adaptations to marine life and the obligate exposure to extreme pressure changes as diving mammals.

First description of a lesion in the upper digestive mucosa associated with a novel gammaherpesvirus in a striped dolphin (Stenella coeruleoalba) stranded in the Western Mediterranean Sea.

BACKGROUND: A wide variety of lesions have been associated with herpesvirus in cetaceans. However, descriptions of herpesvirus infections in the digestive system of cetaceans are scarce. CASE REPORT: A young female striped dolphin stranded in the Valencian Community (Spain) on the 6th August 2021. The animal showed external macroscopic lesions suggestive of an aggressive interaction with bottlenose dolphins (rake marks in the epidermis). Internally, the main findings included congestion of the central nervous system and multiple, well-defined, whitish, irregularly shaped, proliferative lesions on the oropharyngeal and laryngopharyngeal mucosa. Histopathology revealed lymphoplasmacytic and histiocytic meningoencephalitis, consistent with neuro brucellosis. The oropharyngeal and laryngopharyngeal plaques were comprised histologically of focally extensive epithelial hyperplasia. As part of the health surveillance program tissue samples were tested for cetacean morbillivirus using a real-time reverse transcription-PCR, for Brucella spp. using a real-time PCR, and for herpesvirus using a conventional nested PCR. All samples were negative for cetacean morbillivirus; molecular positivity for Brucella spp. was obtained in pharyngeal tonsils and cerebrospinal fluid; herpesvirus was detected in a proliferative lesion in the upper digestive mucosa. Phylogenetic analysis showed that the herpesvirus sequence was included in the Gammaherpesvirinae subfamily. This novel sequence showed the greatest identity with other Herpesvirus sequences detected in skin, pharyngeal and genital lesions in five different species. CONCLUSIONS: To the best of the authors' knowledge, this is the first report of a proliferative lesion in the upper digestive mucosa associated with gammaherpesvirus posititvity in a striped dolphin (Stenella coeruleoalba).

New insights into risk variables associated with gas embolism in loggerhead sea turtles (Caretta caretta) caught in trawls and gillnets.

Tissue and blood gas embolism (GE) associated with fisheries bycatch are likely a widespread, yet underestimated, cause of sea turtle mortality. Here, we evaluated risk factors associated with tissue and blood GE in loggerhead turtles caught incidentally by trawl and gillnet fisheries on the Valencian coastline of Spain. Of 413 turtles (303 caught by trawl, 110 by gillnet fisheries), 54% (n = 222) exhibited GE. For sea turtles caught in trawls, the probability and severity of GE increased with trawl depth and turtle body mass. In addition, trawl depth and the GE score together explained the probability of mortality (P[mortality]) following recompression therapy. Specifically, a turtle with a GE score of 3 caught in a trawl deployed at 110 m had a P[mortality] of ~50%. For turtles caught in gillnets, no risk variables were significantly correlated with either the P[GE] or GE score. However, gillnet depth or GE score, separately, explained P[mortality], and a turtle caught at 45 m or with a GE score between 3 and 4 had a P[mortality] of 50%. Differences in the fishery characteristics precluded direct comparison of GE risk and mortality between these gear types. Although P[mortality] is expected to be significantly higher in untreated turtles released at sea, our findings can improve estimates of sea turtle mortality associated with trawls and gillnets, and help guide associate conservation efforts.

Balaenophilus manatorum in Debilitated and Bycatch-Derived Loggerhead Sea Turtles Caretta caretta from Northwestern Adriatic Sea.

Balenophilus manatorum (Copepoda: Harpaticoida) is one of the few components of the epibiontic fauna of Caretta caretta that show a "true" parasitic association with their host. From rrosive to ulcerative cutaneous lesions may seldom appear as a consequence of the copepod feeding on keratin on turtles' skin. Debilitating Turtle Syndrome (DTS) is the final outcome of a chronic insufficient assumption of nutrients, generally occurring with the impairment of immune functions and high epibiota burdens. In this survey, the presence of B. manatorum in C. caretta from the Northwestern Adriatic Sea was investigated and the relation between infection indices and the co-occurrence of DTS was studied. Clinical examination was performed at the time of rescue, including routine hematological assessment; external parasites were isolated mechanically from turtles' skin and morphologically identified through observation with an optic microscope and SEM. Ten turtles were classified as affected by DTS, all of them being small juveniles with typical clinical and clinicopathological presentation. A higher prevalence, abundance, and density of infection were found in turtles affected by the syndrome. The presence of massive skin coverage by the burrowing barnacle Pletylepas hexastylos prevented a proper evaluation of the pathology associated with B. manatorum in turtles affected by DTS. In any event, eventual skin damages caused by the parasite may represent a port of entry for secondary infections in such immunocompromised animals. Therefore, infection by B. manatorum should not go overlooked in debilitated turtles and should be opportunely treated.

Absence of atrial smooth muscle in the heart of the loggerhead sea turtle (Caretta caretta): a re-evaluation of its role in diving physiology.

Contraction of atrial smooth muscle in the hearts of semi-aquatic emydid turtles regulates ventricular filling, and it has been proposed that it could regulate stroke volume during characteristic rapid transitions in cardiac output associated with diving. For this hypothesis to be supported, atrial smooth muscle should be widely distributed in diving Testudines. To further understand the putative function and evolutionary significance of endocardial smooth muscle in Testudines, we studied the hearts of loggerhead sea turtles, Caretta caretta (n=7), using immunohistochemistry and histology. Surprisingly, we found no evidence of prominent atrial smooth muscle in C. caretta. However, smooth muscle was readily identified in the sinus venosus. Our results suggest that atrial smooth muscle does not contribute to the diving capabilities of C. caretta, indicating that the possible roles of smooth muscle in emydid turtle hearts require a re-evaluation. In sea turtles, the sinus venosus may instead contribute to regulate cardiac filling.

Satellite-tracking dataset of loggerhead sea turtles tracked from western Mediterranean.

We provide the raw data of 44 satellite-tracked loggerhead sea turtles from different life-stages collected between 2016 and 2018. Depending on life-stage and tag availability a different satellite tag was attached to the loggerhead carapace. Location data were collected using the Argos system. We made publically available for the first time in the Mediterranean: (i) the satellite-tracking data for 17 one-year-aged post-hatchlings of loggerhead sea turtle came from two nests laid on the Mediterranean Spanish coast; (ii) the satellite-tracking data for 4 loggerhead nesting females collected from nesting events occurred on the Spanish Mediterranean coast. Besides, another 23 juvenile and adult loggerhead sea turtles were monitored and their data were made also available. Our dataset provide the turtle identity name or number, the date and coordinates of the location data, and the Argos location class associated. Our data contribute to the knowledge about the spatial use of the loggerhead sea turtle in the Mediterranean Sea and could be used in further analysis regarding habitat use and dispersal of this species in the Mediterranean Sea. Specially, nesting females and post-hatchlings data contribute to shed light about these life-stages related to nesting events out of the usual nesting range in the western Mediterranean, for which data still remains scarce. Also, data could be compared to further similar research in satellite-tracking loggerhead sea turtles in the Mediterranean basin.

Systematic Determination of Herpesvirus in Free-Ranging Cetaceans Stranded in the Western Mediterranean: Tissue Tropism and Associated Lesions.

The monitoring of herpesvirus infection provides useful information when assessing marine mammals' health. This paper shows the prevalence of herpesvirus infection (80.85%) in 47 cetaceans stranded on the coast of the Valencian Community, Spain. Of the 966 tissues evaluated, 121 tested positive when employing nested-PCR (12.53%). The largest proportion of herpesvirus-positive tissue samples was in the reproductive system, nervous system, and tegument. Herpesvirus was more prevalent in females, juveniles, and calves. More than half the DNA PCR positive tissues contained herpesvirus RNA, indicating the presence of actively replicating virus. This RNA was most frequently found in neonates. Fourteen unique sequences were identified. Most amplified sequences belonged to the Gammaherpesvirinae subfamily, but a greater variation was found in Alphaherpesvirinae sequences. This is the first report of systematic herpesvirus DNA and RNA determination in free-ranging cetaceans. Nine (19.14%) were infected with cetacean morbillivirus and all of them (100%) were coinfected with herpesvirus. Lesions similar to those caused by herpesvirus in other species were observed, mainly in the skin, upper digestive tract, genitalia, and central nervous system. Other lesions were also attributable to concomitant etiologies or were nonspecific. It is necessary to investigate the possible role of herpesvirus infection in those cases.

Pathological findings and husbandry management in captive Chrysaora spp. medusae affected by umbrellar ulcerative lesions.

During a 12 month period, a group of 14 medusa-stage jellies of the genus Chrysaora, including Pacific sea nettle (Chrysaora fuscescens, n = 11) and Japanese sea nettle (Chrysaora pacifica, n = 3), that were maintained in a public aquarium developed progressive ulcerative umbrellar lesions. In 6 cases (42.9%), ulceration was deep, transmural, and perforated through the mesoglea and subumbrella. In 6 cases (42.9%), ciliated protozoa histomorphologically consistent with scuticociliates were observed in the mesoglea and gastrovascular cavity. In 2 cases (14.3%), commensal dinoflagellates (zooxanthellae) were in the mesoglea and in the cytoplasm of the scuticociliates. During this period, water quality parameters including temperature [°C], pH, oxidation-reduction potential (ORP) [mV], salinity [psu], dissolved oxygen [%], ammonia (NH3), and nitrite(NO2) levels were monitored daily or weekly. The main water quality abnormalities were increased NO2 and pH levels above recommended reference ranges for C. fuscescens and elevated temperature above recommended reference ranges for C. pacifica tank. After correction of water quality parameters, apparent improvement of jellies was observed. In this case, environmental factors were considered the most likely predisposing factors for the development of ulcerative lesions, and ciliated protozoa were considered secondary rather than primary pathogens.

Effects of 3 Tesla magnetic resonance imaging exposure on the behavior and orientation of homing pigeons Columba livia domestica.

Homing pigeons (Columba livia domestica) were used to test whether clinical magnetic resonance (MR) imaging disrupts orientation of animals that sense the earth's magnetic field. Thirty young pigeons were randomly separated into three groups (n = 10/group). Two groups were anaesthetized and exposed to either a constant (no sequence) or a varying (gradient echo and echo planar sequences) magnetic field within a 3 Tesla MR unit for 15 minutes. The control group was not exposed to the MR field but shared all other aspects of the procedure. One day later, animals were released from a site they had never visited, 15 km from the home loft. Three weeks after the procedure, animals were released from a different unfamiliar site 30 km from the loft. Measured variables included the time to disappear from sight (seconds), vanishing bearing (angle), and the time interval from release to entering the home loft (hours). On first release, the group exposed to varying field gradients during image acquisition using 2 different standard sequences showed more variability in the vanishing bearing compared to the other groups (p = 0.0003 compared to control group), suggesting interference with orientation. Other measures did not show significant differences between groups. On second release, there were no significant differences between groups. Our results on homing pigeons show that regular clinical MR imaging exposure may temporarily affect the orientation of species that have magnetoreception capabilities. If exposure to MR imaging disrupted processes that are not specific to magnetoreception, then it may affect other species and other capabilities as well.

Disease risk analysis in sea turtles: A baseline study to inform conservation efforts.

The impact of a range of different threats has resulted in the listing of six out of seven sea turtle species on the IUCN Red List of endangered species. Disease risk analysis (DRA) tools are designed to provide objective, repeatable and documented assessment of the disease risks for a population and measures to reduce these risks through management options. To the best of our knowledge, DRAs have not previously been published for sea turtles, although disease is reported to contribute to sea turtle population decline. Here, a comprehensive list of health hazards is provided for all seven species of sea turtles. The possible risk these hazards pose to the health of sea turtles were assessed and "One Health" aspects of interacting with sea turtles were also investigated. The risk assessment was undertaken in collaboration with more than 30 experts in the field including veterinarians, microbiologists, social scientists, epidemiologists and stakeholders, in the form of two international workshops and one local workshop. The general finding of the DRA was the distinct lack of knowledge regarding a link between the presence of pathogens and diseases manifestation in sea turtles. A higher rate of disease in immunocompromised individuals was repeatedly reported and a possible link between immunosuppression and environmental contaminants as a result of anthropogenic influences was suggested. Society based conservation initiatives and as a result the cultural and social aspect of interacting with sea turtles appeared to need more attention and research. A risk management workshop was carried out to acquire the insights of local policy makers about management options for the risks relevant to Queensland and the options were evaluated considering their feasibility and effectiveness. The sea turtle DRA presented here, is a structured guide for future risk assessments to be used in specific scenarios such as translocation and head-starting programs.

Alpha- and gammaherpesviruses in stranded striped dolphins (Stenella coeruleoalba) from Spain: first molecular detection of gammaherpesvirus infection in central nervous system of odontocetes.

BACKGROUND: Herpesvirus infections in cetaceans have always been attributed to the Alphaherpesvirinae and Gammaherpesvirinae subfamilies. To date, gammaherpesviruses have not been reported in the central nervous system of odontocetes. CASE PRESENTATION: A mass stranding of 14 striped dolphins (Stenella coeruleoalba) occurred in Cantabria (Spain) on 18th May 2019. Tissue samples were collected and tested for herpesvirus using nested polymerase chain reaction (PCR), and for cetacean morbillivirus using reverse transcription-PCR. Cetacean morbillivirus was not detected in any of the animals, while gammaherpesvirus was detected in nine male and one female dolphins. Three of these males were coinfected by alphaherpesviruses. Alphaherpesvirus sequences were detected in the cerebrum, spinal cord and tracheobronchial lymph node, while gammaherpesvirus sequences were detected in the cerebrum, cerebellum, spinal cord, pharyngeal tonsils, mesenteric lymph node, tracheobronchial lymph node, lung, skin and penile mucosa. Macroscopic and histopathological post-mortem examinations did not unveil the potential cause of the mass stranding event or any evidence of severe infectious disease in the dolphins. The only observed lesions that may be associated with herpesvirus were three cases of balanitis and one penile papilloma. CONCLUSIONS: To the authors' knowledge, this is the first report of gammaherpesvirus infection in the central nervous system of odontocete cetaceans. This raises new questions for future studies about how gammaherpesviruses reach the central nervous system and how infection manifests clinically.

Deciphering function of the pulmonary arterial sphincters in loggerhead sea turtles (Caretta caretta).

To provide new insight into the pathophysiological mechanisms underlying gas emboli (GE) in bycaught loggerhead sea turtles (Caretta caretta), we investigated the vasoactive characteristics of the pulmonary and systemic arteries, and the lung parenchyma (LP). Tissues were opportunistically excised from recently dead animals for in vitro studies of vasoactive responses to four different neurotransmitters: acetylcholine (ACh; parasympathetic), serotonin (5HT), adrenaline (Adr; sympathetic) and histamine. The significant amount of smooth muscle in the LP contracted in response to ACh, Adr and histamine. The intrapulmonary and systemic arteries contracted under both parasympathetic and sympathetic stimulation and when exposed to 5HT. However, proximal extrapulmonary arterial (PEPA) sections contracted in response to ACh and 5HT, whereas Adr caused relaxation. In sea turtles, the relaxation in the pulmonary artery was particularly pronounced at the level of the pulmonary artery sphincter (PASp), where the vessel wall was highly muscular. For comparison, we also studied tissue response in freshwater sliders turtles (Trachemys scripta elegans). Both PEPA and LP from freshwater sliders contracted in response to 5HT, ACh and also Adr. We propose that in sea turtles, the dive response (parasympathetic tone) constricts the PEPA, LP and PASp, causing a pulmonary shunt and limiting gas uptake at depth, which reduces the risk of GE during long and deep dives. Elevated sympathetic tone caused by forced submersion during entanglement with fishing gear increases the pulmonary blood flow causing an increase in N2 uptake, potentially leading to the formation of blood and tissue GE at the surface. These findings provide potential physiological and anatomical explanations on how these animals have evolved a cardiac shunt pattern that regulates gas exchange during deep and prolonged diving.

Impact of gas emboli and hyperbaric treatment on respiratory function of loggerhead sea turtles (Caretta caretta).

Fisheries interactions are the most serious threats for sea turtle populations. Despite the existence of some rescue centres providing post-traumatic care and rehabilitation, adequate treatment is hampered by the lack of understanding of the problems incurred while turtles remain entrapped in fishing gears. Recently it was shown that bycaught loggerhead sea turtles (Caretta caretta) could experience formation of gas emboli (GE) and develop decompression sickness (DCS) after trawl and gillnet interaction. This condition could be reversed by hyperbaric O2 treatment (HBOT). The goal of this study was to assess how GE alters respiratory function in bycaught turtles before recompression therapy and measure the improvement after this treatment. Specifically, we assessed the effect of DCS on breath duration, expiratory and inspiratory flow and tidal volume (VT), and the effectiveness of HBOT to improve these parameters. HBOT significantly increased respiratory flows by 32-45% while VT increased by 33-35% immediately after HBOT. Repeated lung function testing indicated a temporal increase in both respiratory flow and VT for all bycaught turtles, but the changes were smaller than those seen immediately following HBOT. The current study suggests that respiratory function is significantly compromised in bycaught turtles with GE and that HBOT effectively restores lung function. Lung function testing may provide a novel means to help diagnose the presence of GE, be used to assess treatment efficacy, and contribute to sea turtle conservation efforts.

Supplemental feeding and other anthropogenic threats to green turtles (Chelonia mydas) in the Canary Islands.

Green turtles are found in the waters of the Canary Islands but little is known about the ecology and anthropogenic pressures that threaten them. Our results have revealed that juvenile green turtles, ranging in curve carapace length from 26.9-81.0cm, are regularly found in the archipelago and originate from rookeries in both the eastern and western Atlantic. Photo-identification and satellite tracking showed high levels of site fidelity to coastal foraging grounds associated with seagrass meadows, but stable isotope analysis indicated animal-based omnivorous diets after settlement on the continental shelf, with no increase in the consumption of macrophytes as the turtles grew. Most turtles exhibited high levels of some blood biochemical markers associated with a high consumption of proteins and fat. In addition, we determined levels of some organic and inorganic pollutants. Supplemental feeding may also contribute to explain the high prevalence of hooking and boat strikes in the green turtles brought to wildlife rescue centers as compared with loggerhead turtles. Regulatory measures and surveillance should be urgently implemented in order to improve the status of the species in the archipelago.

Defining risk variables causing gas embolism in loggerhead sea turtles (Caretta caretta) caught in trawls and gillnets.

Incidental capture, or 'bycatch' in fishing gear is a major global threat to sea turtle populations. A recent study showed that underwater entrapment in fishing gear followed by rapid decompression may cause gas bubble formation within the blood stream (embolism) and tissues leading to organ injury, impairment, and even mortality in some bycaught individuals. We analyzed data from 128 capture events using logistic and ordinal regression to examine risk factors associated with gas embolism in sea turtles captured in trawls and gillnets. Likelihood of fatal decompression increases with increasing depth of gear deployment. A direct relationship was found between depth, risk and severity of embolism, which has not been previously demonstrated in any breath-hold diving species. For the trawl fishery in this study, an average trawl depth of 65 m was estimated to result in 50% mortality in by-caught turtles throughout the year. This finding is critical for a more accurate estimation of sea turtle mortality rates resulting from different fisheries and for devising efforts to avoid or minimize the harmful effects of capture.

To Swim or Not to Swim: Potential Transmission of Balaenophilus manatorum (Copepoda: Harpacticoida) in Marine Turtles.

Species of Balaenophilus are the only harpacticoid copepods that exhibit a widespread, obligate association with vertebrates, i.e., B. unisetus with whales and B. manatorum with marine turtles and manatees. In the western Mediterranean, juveniles of the loggerhead sea turtle, Caretta caretta are the only available hosts for B. manatorum, which has been found occurring at high prevalence (>80%) on them. A key question is how these epibionts are transmitted from host to host. We investigated this issue based on experiments with live specimens of B. manatorum that were cultured with turtle skin. Specimens were obtained from head-started hatchlings of C. caretta from the western Mediterranean. Hatched nauplii crawled only on rough substrates and lacked the ability to swim. Only copepodites IV and V, and adults, were able to perform directional swimming. Legs 2, 3 and 4 played a major role in swimming and were only well-developed in these stages. Nauplii reared in wells with turtle skin readily fed on this item. Late copepodites and adults also fed on turtle skin but did not consume other potential food items such as fish skin, baleen plates or planktonic algae. Evidences suggest that the transmission of B. manatorum should rely on hosts' bodily contacts and/or swimming of late developmental stages between spatially close hosts. The possibility of long-ranged dispersal is unlikely for two reasons. First, all developmental stages seem to depend on turtle skin as a food resource. Second, the average clutch size of ovigerous females was small (< 70 eggs) for free-living phases to successfully contact turtles that occur at very low densities (< 0.6 turtles·km-2) in the western Mediterranean. The high prevalence of B. manatorum in loggerhead turtles in this area raises the question whether these turtles have contacts, or tend to closely aggregate, more than is currently believed.

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.

First molecular determination of herpesvirus from two mysticete species stranded in the Mediterranean Sea.

BACKGROUND: Herpesvirus can infect a wide range of animal species: mammals, birds, reptiles, fish, amphibians and bivalves. In marine mammals, several alpha- and gammaherpesvirus have been identified in some cetaceans and pinnipeds species. To date, however, this virus has not been detected in any member of the Balaenoptera genus. CASE PRESENTATION: Herpesvirus was determined by molecular methods in tissue samples from a male fin whale juvenile (Balaenoptera physalus) and a female common minke whale calf (Balaenoptera acutorostrata) stranded on the Mediterranean coast of the Region of Valencia (Spain). Samples of skin and penile mucosa from the fin whale and samples of skin, muscle and central nervous system tissue from the common minke whale tested positive for herpesvirus based on sequences of the DNA polymerase gene. Sequences from fin whale were identical and belonged to the Alphaherpesvirinae subfamily. Only members of the Gammaherpesvirinae subfamily were amplified from the common minke whale, and sequences from the muscle and central nervous system were identical. Sequences in GenBank most closely related to these novel sequences were viruses isolated from other cetacean species, consistent with previous observations that herpesviruses show similar phylogenetic branching as their hosts. CONCLUSIONS: To our knowledge, this is the first molecular determination of herpesvirus in the Balaenoptera genus. It shows that herpesvirus should be included in virological evaluation of these animals.