Historical isolates of dolphin morbillivirus: origin and new data.

Dolphin morbillivirus (DMV) was isolated in striped dolphins Stenella coeruleoalba from the Mediterranean Sea stranded along the coast of Spain during a lethal epidemic that killed thousands of individuals in 1990-1992. Though some of these isolates (MUC, 16A and the reference strain) have been extensively characterised, details on their origin were not reported in the literature, and records for these isolates are often difficult to trace and are, sometimes, erroneous. Here, we provide unpublished biological and histopathological data for these isolates, summarize the literature on their characterization and make suggestions for future studies.

From development to taxonomy: the case of Sciaenacotyle pancerii (Monogenea: Microcotylidae) in the Mediterranean meagre.

The microcotylid Sciaenacotyle pancerii is a pathogenic monogenean infecting Argyrosomus regius, a candidate for species diversification in the Mediterranean aquaculture. Life-history stages of S. pancerii commonly co-occur in field infections, but to date, morphological data have only been provided for oncomiracidia and adults although identifying life-history stages can be useful in infection management. A total of 114 specimens of S. pancerii were analysed to characterize the developmental events and to assess morphological and morphometric variations before and after maturity. The post-larval development of S. pancerii is characterized by: expansion and bifurcation of the gut, loss of the larval haptor, protandrous development of the genitalia and vitellaria formation. The size variability of larval hooks, hamuli and germanium of S. pancerii is firstly reported and dimensional ranges of parasite body, haptor, testes, posteriormost clamps and eggs are widened. The size of most of the diagnostic features of S. pancerii significantly increases after parasite maturity and therefore, only those specimens with more than 116 clamps should be considered for minimising development-related variability in size. The high number of clamps, their fast development and the asymmetry in their size and arrangement suggest that S. pancerii may use a mixed attachment strategy between the closely related microcotylids and heteraxinids. This combination of features may be host related and linked to the gill structure of the sciaenid fish and the phylogenetic position of the genus Sciaenacotyle; distant from other microcotylids while close to heteraxinid species.

Cardicola mediterraneus n. sp. (Trematoda, Aporocotylidae): a new species infecting the gilthead seabream, Sparus aurata L., from the Western Mediterranean Sea.

The genus Cardicola Short, 1953 has the highest number of species within the family Aporocotylidae (Trematoda: Digenea). Five Cardicola species have been reported to date in the Mediterranean Sea, one of them in the gilthead seabream Sparus aurata L. Analyses of infected S. aurata recovered from cultured fish off Sardinia (Italy) and from wild and cultured fish off the Levantine coast (Southeastern Spain) have revealed the presence of two species identified as Cardicola aurata Holzer, Montero, Repullés, Nolan, Sitjà-Bobadilla, Álvarez-Pellitero, Zarza and Raga, 2008 and Cardicola mediterraneus n. sp.. New morphological and molecular data are provided for both species. Features of C. aurata specimens differ slightly from those of the original description of the species, the most important differences being the longer extension of the metraterm and the central and posterior position of the female genital pore. Cardicola mediterraneus n. sp. can be easily distinguished from other Cardicola species by two unique specific characters: (i) the very unequal posterior caeca length and (ii) the shape of the testis, deeply notched at the anterior extremity. Cardicola spp. from sparids occupy a basal phylogenetic position respect the other congeneric species. The genus Cardicola has a complex taxonomy and shows high intrageneric differences for both 28S and ITS2 rDNAs, similar to the intergeneric differences among other aporocotylid genera, suggesting that it could be split. The presence of two Cardicola species could hamper treatment design and application; thus, data discriminating species herein reported can improve the infection management in fish farms.

Sex-specific impact of inbreeding on pathogen load in the striped dolphin.

The impact of inbreeding on fitness has been widely studied and provides consequential inference about adaptive potential and the impact on survival for reduced and fragmented natural populations. Correlations between heterozygosity and fitness are common in the literature, but they rarely inform about the likely mechanisms. Here, we investigate a pathology with a clear impact on health in striped dolphin hosts (a nematode infection that compromises lung function). Dolphins varied with respect to their parasite burden of this highly pathogenic lung nematode (Skrjabinalius guevarai). Genetic diversity revealed by high-resolution restriction-associated DNA (43 018 RADseq single nucleotide polymorphisms) analyses showed a clear association between heterozygosity and pathogen load, but only for female dolphins, for which the more heterozygous individuals had lower Sk. guevarai burden. One locus identified by RADseq was a strong outlier in association with parasite load (heterozygous in all uninfected females, homozygous for 94% of infected females), found in an intron of the citron rho-interacting serine/threonine kinase locus (associated with milk production in mammals). Allelic variation at the Class II major histocompatability complex DQB locus was also assessed and found to be associated with both regional variation and with pathogen load. Both sex specificity and the identification of associating functional loci provide insight into the mechanisms by which more inbred individuals may be more susceptible to the infection of this parasite. This provides important insight towards our understanding of the impact of inbreeding in natural populations, relevant to both evolutionary and practical conservation considerations.

Cranial crassicaudiasis in two coastal dolphin species from South Africa is predominantly a disease of immature individuals.

Crassicauda spp. (Nematoda) infest the cranial sinuses of several odontocetes, causing diagnostic trabecular osteolytic lesions. We examined skulls of 77 Indian Ocean humpback dolphins Sousa plumbea and 69 Indo-Pacific bottlenose dolphins Tursiops aduncus, caught in bather-protecting nets off KwaZulu-Natal (KZN) from 1970-2017, and skulls of 6 S. plumbea stranded along the southern Cape coast in South Africa from 1963-2002. Prevalence of cranial crassicaudiasis was evaluated according to sex and cranial maturity. Overall, prevalence in S. plumbea and T. aduncus taken off KZN was 13 and 31.9%, respectively. Parasitosis variably affected 1 or more cranial bones (frontal, pterygoid, maxillary and sphenoid). No significant difference was found by gender for either species, allowing sexes to be pooled. However, there was a significant difference in lesion prevalence by age, with immature T. aduncus 4.6 times more likely affected than adults, while for S. plumbea, the difference was 6.5-fold. As severe osteolytic lesions are unlikely to heal without trace, we propose that infection is more likely to have a fatal outcome for immature dolphins, possibly because of incomplete bone development, lower immune competence in clearing parasites or an over-exuberant inflammatory response in concert with parasitic enzymatic erosion. Cranial osteolysis was not observed in mature males (18 S. plumbea, 21 T. aduncus), suggesting potential cohort-linked immune-mediated resistance to infestation. Crassicauda spp. may play a role in the natural mortality of S. plumbea and T. aduncus, but the pathogenesis and population level impact remain unknown.

Causes of cetacean stranding and death on the Catalonian coast (western Mediterranean Sea), 2012-2019.

The causes of cetacean stranding and death along the Catalan coast between 2012 and 2019 were systematically investigated. Necropsies and detailed pathological investigations were performed on 89 well-preserved stranded cetaceans, including 72 striped dolphins Stenella coeruleoalba, 9 Risso's dolphins Grampus griseus, 5 bottlenose dolphins Tursiops truncatus, 1 common dolphin Delphinus delphis, 1 Cuvier's beaked whale Ziphius cavirostris and 1 fin whale Balaenoptera physalus. The cause of death was determined for 89.9% of the stranded cetaceans. Fisheries interaction was the most frequent cause of death in striped dolphins (27.8%) and bottlenose dolphins (60%). Cetacean morbillivirus (CeMV) was detected on the Catalan coast from 2016 to 2017, causing systemic disease and death in 8 of the 72 (11.1%) striped dolphins. Chronic CeMV infection of the central nervous system was observed from 2018-2019 in a further 5 striped dolphins. Thus, acute and chronic CeMV disease caused mortality in 18% of striped dolphins and 14.6% of all 89 cetaceans. Brucella ceti was isolated in 6 striped dolphins and 1 bottlenose dolphin with typical brucellosis lesions and in 1 striped dolphin with systemic CeMV. Sinusitis due to severe infestation by the nematode parasite Crassicauda grampicola caused the death of 4 out of 6 adult Risso's dolphins. Maternal separation, in some cases complicated with septicemia, was a frequent cause of death in 13 of 14 calves. Other less common causes of death were encephalomalacia of unknown origin, septicemia, peritonitis due to gastric perforation by parasites and hepatitis caused by Sarcocystis spp.

The taxonomic status of Skrjabinalius guevarai Gallego & Selva, 1979 (Nematoda: Pseudaliidae) and the synonymy of Skrjabinalius Delyamure, 1942 and Halocercus Baylis & Daubney, 1925.

Within the cetacean lungworm family Pseudaliidae Raillet & Henry, 1909, the distinction between the two genera of the subfamily Halocercinae Delamure, 1952, i.e. Halocercus Baylis & Daubney, 1925 and Skrjabinalius Delyamure, 1942, is principally based on the structure and shape of the male copulatory bursa. In species of Halocercus, the bursa is unlobed, whereas in the two species included in Skrjabinalius, S. cryptocephalus Delyamure, 1942 and Skrjabinalius guevarai Gallego & Selva, 1979, the bursa is described as clearly lobed. During a parasitological analysis of cetaceans from the Spanish Mediterranean, we collected a number of specimens of S. guevarai with variable levels of bursal lobulation, including individuals with unlobed bursae. Examination of voucher specimens of the type-species of Halocercus, H. delphini Baylis & Daubney, 1925, collected from cetaceans in the North-East Atlantic revealed the same variations in bursal shape, and the same arrangement of bursal rays and papillae. A morphometric comparison did not reveal substantial differences between both species. Moreover, Maximum Likelihood and Bayesian Inference analyses of the second internal transcribed spacer (ITS2) sequences of 11 specimens of S. guevarai and one of H. delphini grouped both species together, regardless of bursal shape, in a highly supported clade within the Pseudaliidae. Accordingly, we consider S. guevarai as a junior synonym of H. delphini. The great variability found in bursal lobulation in the type-species of Halocercus invalidates the use of this trait as a genus-level diagnostic character and, therefore, Skrjabinalius should also be considered synonymous with Halocercus.

Taxonomic status and epidemiology of the mesoparasitic copepod Pennella balaenoptera in cetaceans from the western Mediterranean.

Pennella balaenoptera is a mesoparasitic copepod that has been reported in at least 17 cetacean species. Subtle morphological differences in the first antennae of adult females have been used to discriminate this species from P. filosa, a species infecting fishes. Other morphological traits are unreliable because of their high plasticity, and no molecular data are available to confirm the taxonomic status of P. balaenoptera as an independent species. We found no consistent morphological differences of the first antennae between P. balaenoptera and P. filosa collected from cetaceans and fish in the western Mediterranean. Molecular data on the mitochondrial cytochrome oxidase subunit I failed to show reciprocal monophyly for the 2 species, and nucleotide divergence between them was low (mean ± SD [range]: 4.1 ± 0.006% [0.5-8.9]). Thus, P. balaenoptera and P. filosa are considered conspecific. We also obtained data on infection parameters of P. balaenoptera based on 450 individuals of 6 cetacean species stranded on the Spanish Mediterranean coast between 1980 and 2017. Prevalence was significantly lowest in the most coastal species, the bottlenose dolphin Tursiops truncatus (3.6%) and highest in the most oceanic species, Cuvier's beaked whale Ziphius cavirostris (100%). This suggests that the life cycle of P. balaenoptera is primarily oceanic. Interestingly, P. filosa also occurs in the oceanic realm infecting large fishes. This ecological similarity further supports the hypothesis that P. balaenoptera and P. filosa are conspecific.

Skoulekia erythrini n. sp. (Digenea: Aporocotylidae): a parasite of Pagellus erythrinus (L.) (Perciformes: Sparidae) from the western Mediterranean with an amendment of the generic diagnosis.

A new aporocotylid, Skoulekia erythrini n. sp., is described from the heart, cephalic kidney and gill blood vessels of the common pandora Pagellus erythrinus (L.) collected from several localities of the western Mediterranean off Spain. The new species differs from the type- and only species of the genus, S. meningialis Alama-Bermejo, Montero, Raga & Holzer, 2011 in possessing a symmetrical body (vs laterally curved), short tegumental spines (3-4 vs 7-10 µm) without hooked ends, diffuse (vs conspicuous) oesophageal gland-cells, a relatively longer oesophagus, a testis that is shorter in relation to body length and a much smaller seminal vesicle (17-34 × 10-26 vs 33-101 × 27-97 µm). Elongated ellipsoidal eggs of S. erythrini n. sp. were found trapped in gill vessels in histological sections. Phylogenetic analyses based on partial 28S rDNA and ITS2 sequences supported the placement of the new species within Skoulekia and the close relationships of this genus with Psettarium Goto & Ozaki, 1929 and Pearsonellum Overstreet & Køie, 1986. Skoulekia meningialis is described from a new sparid host, Diplodus puntazzo (Walbaum), collected off Santa Pola, Spain. The new morphological data for the two Skoulekia spp. and a re-examination of three paratypes of S. meningialis pinpointed features amending both the description of S. meningialis and the generic diagnosis of Skoulekia.

Trematode maturation patterns in a migratory snail host: What happens during upshore residency in a Mediterranean lagoon?

Maturation of trematode larval stages is expected to be temporally and spatially adapted to maximise the encounter with the adequate downstream host, i.e. the host, which will be infected by this parasite stage. Since studies on intramolluscan parasite maturation are scarce but important in the context of parasite transmission, the larval development inside sporocysts was monitored during upshore residency of the snail host Gibbula adansonii (Trochidae), i.e., from March to May (2011 and 2013), when these snails temporarily reside in the intertidal habitat of a Western Mediterranean lagoon (40° 37' 35″ N, 0° 44' 31″ E, Spain). Data on the relative quantity of different maturation stages of Cainocreadium labracis and Macvicaria obovata (Opecoelidae) parasitising the G. adansonii as well as on snail and sporocyst size were explored using linear models and linear mixed models. The effect of the trematodes on snail growth was shown to be species-specific, with snail and sporocyst size acting as proxies of the reproductive capacity of M. obovata but not that of C. labracis. The number of cercarial embryos and germinal balls did not show monthly variation in either parasite species, but a higher number of mature stages and the highest maturity index was found in April. Hence, during the snail's limited spawning-related presence in the upshore waters of the lagoon, continuous production and output of infectious cercariae was observed, which indicates a link between larval maturation and snail migration. The synchronization of snails, mature parasite transmission stages and downstream hosts in time and space guarantees a successful completion of the life cycle.

Same host, same lagoon, different transmission pathways: effects of exogenous factors on larval emergence in two marine digenean parasites.

Due to their shallow and confined nature, lagoons provide excellent conditions for the transmission of digenean trematode parasites that require two or more intermediate hosts for the completion of their complex life cycles. However, these unstable environments are characterised by an internal heterogeneity and a large variation of a range of abiotic variables. We conducted a series of experiments in a comparative framework to assess the effect of a number of exogenous factors known to exhibit marked fluctuations in the lagoonal environment, i.e. temperature, salinity, water level and photoperiod, on larval emergence of two sympatric parasites, Cainocreadium labracis and Macvicaria obovata, which share the snail intermediate host, Gibbula adansonii, and a sit-and-wait downstream host-finding strategy. Our results demonstrated contrasting patterns and rates of larval emergence indicating an overall differential response of the two species to the variation of the environmental factors. Cercariae of M. obovata exhibited increased emergence rates at elevated temperature (with a sharp increase at 15 °C), decreased salinity (35-25 practical salinity units (psu)) and low water levels, whereas larval emergence of C. labracis was unaffected by the experimental variation in temperature and water level and showed decreased rates at decreased salinity levels (35-25 psu). The differential impact of the variable environmental conditions indicates the complexity of the patterns of exogenous control modifying parasite transmission and abundance in the lagoonal environment. Most importantly, the contrasting rhythms of larval emergence indicate endogenous control associated with the different transmission pathways of the two opecoelid digeneans.

A revision of the genus Neogrubea Dillon & Hargis, 1968 (Monogenea: Mazocraeidae): new morphological and molecular data from off the Patagonian coast of Argentina.

The genus Neogrubea Dillon & Hargis, 1968 (syn. Asymmetria Suriano, 1975) is revised based on examination of type- and voucher material, and from new specimens collected from the gills of Seriolella porosa Guichenot and Stromateus brasiliensis Fowler from off Patagonia, Argentina. Morphological comparisons based on light and scanning electron microscopy and molecular data (partial SSU and LSU rDNA sequences) of the monogeneans from off Patagonia suggest that N. seriolellae Dillon & Hargis, 1968 (syns N. stromateae Gibson, 1976, A. asymmetria Suriano, 1975 and A. platensis Rey & Meneses, 1985) is currently the only species of the genus. Neogrubea soni Evdokimova, 1969 is considered a species inquirenda. An emended diagnosis of Neogrubea is presented, and new host and locality records for N. seriolellae are given in detail. Morphological characters of the members of the mazocraeid subfamily Grubeinae Price, 1961 are also discussed.

Molecular phylogeny of the Pseudaliidae (Nematoda) and the origin of associations between lungworms and marine mammals.

Metastrongyloid lungworms from the family Pseudaliidae infect the lungs and cranial sinuses of cetaceans worldwide, except Stenuroides herpestis, which exhibits a striking terrestrial association with the Egyptian mongoose, Herpestes ichneumon. Previous phylogenies of the Metastrongyloidea that included some (2-7) marine species of the Pseudaliidae confirmed that these species are closely related, but also grouped species of Parafilaroides (family Filaroididae) between the members of the Pseudaliidae. In this study we extracted DNA from representatives of all six genera of the Pseudaliidae and amplified the second internal transcribed spacer (ITS2) and cytochrome c oxidase subunit I (cox1) genes in order to investigate the concept of the Pseudaliidae as a monophyletic unit. Three species of Parafilaroides were also included in the analysis. Maximum Likelihood and Bayesian Inference analyses of the concatenated genes resulted in a well-supported clade including the marine pseudaliids, S. herpestis and Parafilaroides spp. These findings validate the status of S. herpestis as a pseudaliid species and support the inclusion of Parafilaroides in the Pseudaliidae. Although males of Parafilaroides spp. lack a copulatory bursa, this trait itself is highly variable in the Pseudaliidae, which include abursate species. Furthermore, life cycles seem to be closely similar between both taxa. When all the available phylogenetic data on the Metastrongyloidea were mapped of onto a phylogeny of the Laurasiatheria, there was a strong suggestion that the Pseudaliidae may have descended from ancestors infecting terrestrial carnivores, with odontocetes being colonized in the marine realm as a result of a host-switching event from pinnipeds through shared fish prey. The origin of the association between S. herpestis and mongooses remains uncertain.

Mapping a brain parasite: occurrence and spatial distribution in fish encephalon.

Parasites, especially brain-encysting trematodes, can have an impact on host behaviour, facilitating the transmission to next host and completion of the life cycle, but insufficient research has been done on whether specific brain regions are targeted. Using Cardiocephaloides longicollis as a laboratory model, the precise distribution of metacercariae in experimentally-infected, wild and farmed fish was mapped. The brain regions targeted by this parasite were explored, also from a histologic perspective, and potential pathogenic effects were evaluated. Experimental infections allowed to reproduce the natural infection intensity of C. longicollis, with four times higher infection intensity at the higher dose (150 vs 50 cercariae). The observed metacercarial distribution, similar among all fish groups, may reflect a trematode species-specific pattern: metacercariae occur with highest density in the optic lobe area (primarily infecting the periventricular gray zone of optic tectum) and the medulla oblongata, whereas other areas such as the olfactory lobes and cerebellar lobes may be occupied when the more frequently invaded parts of the brain were crowded. Mono- and multicysts (i.e. formed either with a single metacercaria, or with 2-25 metacercariae encapsulated together) may be formed depending on the aggregation and timing of metacercariae arrival, with minor host inflammatory response. Larvae of C. longicollis colonizing specific brain areas may have an effect on the functions associated with these areas, which are generally related to sensory and motor functions, but are also related to other host fitness traits such as school maintenance or recognition of predators. The detailed information on the extent and distribution of C. longicollis in fish encephalon sets the ground to understand the effects of brain parasites on fish, but further investigation to establish if C. longicollis, through purely mechanical damage (e.g., occupation, pressure and displacement), has an actual impact on host behaviour remains to be tested under controlled experimental conditions.

Parasite communities in a marine fish indicate ecological recovery from the impacts of the Prestige oil-spill 12-13 years after the disaster.

The Prestige oil-spill (2002) is one of the major marine accidents resulting in contamination of virtually all types of marine habitat along c.900 km of coastline in the North-East Atlantic. Environmental pollution affects parasite populations and communities, both directly and through effects on intermediate and definitive hosts. However, the effects of oil-spills on shelf benthic communities are poorly known. This study addressed the hypothesis of recovery of parasite communities in a marine sparid teleost, the bogue Boops boops (L.) (Teleostei: Sparidae), as indicators of environmental pollution and its effects on benthic/pelagic invertebrate communities in an impacted area off the Galician coast, Spain, 12-13 years after the Prestige oil-spill. Novel data for the metazoan parasite communities collected during 2014-2015 were analysed in association with two unique datasets, one comprising baseline data collected in 2001, one year before the Prestige oil-spill, and one comprising data collected 3-4 years post-spill (2005-2006). Using the taxonomically consistent data on parasites in a series of fish seasonal samples, we found significant differences between the two seasonal post-spill datasets taken at a 9-year time interval (3-4 years and 12-13 years post-spill) in most community metrics and infection parameters of the common species. This was in sharp contrast with the few differences between the long-term post-spill dataset of 2014-2015 and the pre-spill dataset of 2001. Multivariate community similarity analyses confirmed that these differences reflected in the substantial differentiation of parasite community composition and structure of the post-spill communities and the significant homogenisation of communities sampled 12-13 years post-spill and baseline communities. Overall, the novel analyses demonstrated a long-term directional trend in parasite community succession towards ecological recovery irrespective of the natural seasonal variability. These findings suggest longer-term oil-spill impacts on shelf benthic/pelagic invertebrate communities lasting over 10 years.

A severe microsporidian disease in cultured Atlantic Bluefin Tuna (Thunnus thynnus).

One of the most promising aquaculture species is the Atlantic bluefin tuna (Thunnus thynnus) with high market value; disease control is crucial to prevent and reduce mortality and monetary losses. Microsporidia (Fungi) are a potential source of damage to bluefin tuna aquaculture. A new microsporidian species is described from farmed bluefin tunas from the Spanish Mediterranean. This new pathogen is described in a juvenile associated with a highly severe pathology of the visceral cavity. Whitish xenomas from this microsporidian species were mostly located at the caecal mass and ranged from 0.2 to 7.5 mm. Light and transmission electron microscopy of the spores revealed mature spores with an average size of 2.2 × 3.9 µm in size and a polar filament with 13-14 coils arranged in one single layer. Phylogenetic analysis clustered this species with the Glugea spp. clade. The morphological characteristics and molecular comparison confirm that this is a novel microsporidian species, Glugea thunni. The direct life-cycle and the severe pathologies observed makes this parasite a hard risk for bluefin tuna cultures.

Living together but remaining apart: Atlantic and Mediterranean loggerhead sea turtles (Caretta caretta) in shared feeding grounds.

Juvenile loggerhead sea turtles (Caretta caretta) from Atlantic nesting populations migrate into the western Mediterranean, where they share feeding grounds with turtles originating in the Mediterranean. In this scenario, male-mediated gene flow may lead to the homogenization of these distant populations. To test this hypothesis, we genotyped 7 microsatellites from 56 Atlantic individuals sampled from feeding grounds in the western Mediterranean and then compared the observed allele frequencies with published data of 112 individuals from Mediterranean nesting beaches. Mediterranean populations were found to be genetically differentiated from the Atlantic stock reaching the western Mediterranean (F(st) = 0.029, P < 0.001); therefore, the possible mating events between Atlantic and Mediterranean individuals are not sufficient to homogenize these 2 areas. The differentiation observed between these 2 areas demonstrates that microsatellites are sufficiently powerful for mixed stock analysis and that individual assignment (IA) tests can be performed in combination with mitochondrial DNA (mtDNA) analysis. In a set of 197 individuals sampled in western Mediterranean feeding grounds, 87% were robustly assigned to Atlantic or Mediterranean groups with the combined marker, as compared with only 52% with mtDNA alone. These findings provide a new approach for tracking the movements of these oceanic migrants and have strong implications for the conservation of the species.

Post-epizootic chronic dolphin morbillivirus infection in Mediterranean striped dolphins Stenella coeruleoalba.

Dolphin morbillivirus (DMV) has caused 2 epizootics with high mortality rates on the Spanish Mediterranean coast, in 1990 and 2006-07, mainly affecting striped dolphins Stenella coeruleoalba. Following the first epizootic unusual DMV infections affecting only the central nervous system of striped dolphins were found, with histological features similar to subacute sclerosing panencephalitis and old dog encephalitis, the chronic latent localised infections caused by defective forms of measles virus and canine distemper virus, respectively. Between 2008 and 2010, monitoring by microscopic and immunohistochemical (IHC) studies of 118 striped dolphins stranded along Catalonia, the Valencia Region and Andalusia showed similar localised DMV nervous system infections in 25.0, 28.6 and 27.4% of cases, respectively, with no significant differences among regions or sex. The body length of DMV-infected dolphins was statistically greater than that of non-infected dolphins (196.5 vs. 160.5 cm; p < 0.001). Molecular detection of DMV was performed by 2 different RT-PCR techniques amplifying a 429 bp fragment and a 78 bp fragment both within the phosphoprotein (P) gene. The 429 bp RT-PCR results contradicted the IHC-DMV results as only 3 of 6 dolphins with positive IHC-DMV had positive PCR results. All 6 cases were positive with the 78 bp RT-PCR. These findings contraindicate the use of the 429 bp RT-PCR protocol based on the P gene to detect this specific form of DMV. DMV localised nervous infection constitutes the most relevant single cause of stranding and death in Mediterranean striped dolphins in the years following a DMV epizootic, and it might even overwhelm the effects of the epizootic itself, at least in 2007.

Determinants of lungworm specificity in five cetacean species in the western Mediterranean.

BACKGROUND: Current data about Pseudaliidae show contrasting patterns of host specificity between congeneric species. We investigated how both contact and compatibility between hosts and parasites contributed to the patterns of lungworm infection observed in a community of five species of cetaceans in the western Mediterranean. METHODS: The lungs of 119 striped dolphins Stenella coeruleoalba, 18 bottlenose dolphins Tursiops truncatus, 7 Risso's dolphins Grampus griseus, 7 long-finned pilot whales Globicephala melas, and 6 common dolphins Delphinus delphis were analysed for lungworms. Parasites were identified by morphology and analysis of ITS2 sequences using both maximum likelihood and Bayesian inference methods. Body length was used as a proxy for lungworm species fitness in different hosts and compared with Kruskal-Wallis tests. Infection parameters were compared between cetacean species using Fisher's exact tests and Kruskal-Wallis tests. Phylogenetic specificity was explored by collating the overall lungworm species prevalence values in hosts from previous surveys in various localities. To explore the relative importance of vertical and horizontal transmission, Spearman's rank correlation was used to look for an association between host size and lungworm burden. A Mantel test was used to explore the association between lungworm species similarity and prey overlap using dietary data. RESULTS: Halocercus delphini had higher infection levels in striped dolphins and common dolphins; Stenurus ovatus had higher infection levels in bottlenose dolphins; and Stenurus globicephalae had higher infection levels in long-finned pilot whales. These results are congruent with findings on a global scale. Morphometric comparison showed that the larger nematodes were found in the same host species that had the highest parasite burden. Lungworms were found in neonatal striped dolphins and a Risso's dolphin, and there was a weak but significant correlation between host size and parasite burden in striped dolphins and bottlenose dolphins. There was also a weak but significant association between prey overlap and lungworm species similarity. CONCLUSIONS: Data indicate that phylogenetic specificity has an important role in governing host-parasite associations, as indicated by the higher infection levels and larger nematode size in certain hosts. However, diet can also influence infection patterns in these preferred hosts and contribute to less severe infections in other hosts.

Transmission and Predictors of Burden of Lungworms of the Striped Dolphin (Stenella coeruleoalba) in the Western Mediterranean.

Pseudaliid lungworms infect the lungs and sinuses of cetaceans. Information on the life cycle and epidemiology of pseudaliids is very scarce and mostly concerns species that infect coastal or inshore cetaceans. Available evidence indicates that some pseudaliids are vertically transmitted to the host, whereas others are acquired via infected prey. We documented pseudaliid infections in an oceanic cetacean, the striped dolphin (Stenella coeruleoalba) in the western Mediterranean, and investigated the possibilities of vertical vs. horizontal transmission and the potential influence of host body size, sex, and season on infection levels. We found two species of lungworm in 87 dolphins that stranded along the Spanish Mediterranean coast between 1987 and 2018. One or two larvae of Stenurus ovatus were found in three adult dolphins. Larger numbers of larvae and adults of Skrjabinalius guevarai were collected in 51 dolphins, including unweaned calves. These observations suggested that Skrjabinalius guevarai could be vertically transmitted. The abundance of Skrjabinalius guevarai increased significantly with host size, which suggested that it could be trophically transmitted, as well, with larger hosts consuming more infected prey. Infection levels peaked in spring, outside of the calving season, which is likely a reflection of a seasonal shift in dolphin diet. In summary, results indicate that Skrjabinalius guevarai was capable of both vertical and horizontal transmission, but future research should be directed at clarifying the potential mechanics behind transmission and intermediate hosts.