Natalia Fraija-Fernández

Independent host switching events by digenean parasites of cetaceans inferred from ribosomal DNA.

Cetaceans harbour a unique fauna of digeneans whose origin and relationships have sparked considerable debate during recent decades. Disparity in the species reported indicates that they do not share close affinities, but their unusual morphology has made their taxonomic identities and phylogenetic positions uncertain. Here we use sequence data to investigate the phylogenetic relationships of the main species of flukes infecting cetaceans. We sequenced the 18S, 28S and internal transcribed spacer 2 rDNA of digenean species representing all known families reported from cetaceans: Braunina cordiformis (Brauninidae), Ogmogaster antarcticus (Notocotylidae), Pholeter gastrophilus (Heterophyidae), and Campula oblonga, Nasitrema sp. and Oschmarinella rochebruni (Brachycladiidae). The phylogenetic position of the taxa was estimated by Bayesian inference and maximum likelihood incorporating published sequences of 177 species of Digenea. Further Bayesian and maximum likelihood analyses were performed with sequences of 14 Heterophyidae and Opisthorchiidae taxa, incorporating new sequences of P. gastrophilus. Species nominally assigned to the Brachycladiidae formed a clade that was embedded among species of the Acanthocolpidae, thus making the latter family paraphyletic. Braunina cordiformis formed a sister lineage to the Strigeidae and Diplostomidae, whereas O. antarcticus was placed within the Notocotylidae, in agreement with the previous taxonomy of this genus. Similarly, P. gastrophilus was placed within the Heterophyidae as originally described. Our results suggest a paraphyletic relationship between the Heterophyidae and Opisthorchiidae, mirroring the uncertain taxonomic placement of P. gastrophilus, which has been assigned to both families in the past. The digenean families involved are parasites of fish-eating birds and mammals (i.e. Strigeidae, Diplostomidae and Heterophyidae), parasites of marine fish (i.e. Acanthocolpidae) and other herbivorous aquatic birds and mammals (i.e. Notocotylidae). The phylogenetic positions of these taxa indicate that the digenean fauna of cetaceans may have been acquired through independent host-capture events, with two clades showing subsequent diversification exclusively among marine mammals.

Evolutionary relationships between digeneans of the family Brachycladiidae Odhner, 1905 and their marine mammal hosts: A cophylogenetic study.

Cophylogenetic studies examine the congruence between host and parasite phylogenies. There are few studies that quantify the relative contribution of coevolutionary events, i.e. duplication, loss, failure-to-diverge, host-switching and spreading in trophically-transmitted parasites at the marine realm. We addressed this issue in the Brachycladiidae, a cosmopolitan digenean family specific to marine mammals. We used, for the first time, distance-based and event-based methods to explicitly test the coevolutionary events that have shaped the current brachycladiid-marine mammal associations. Parasite phylogeny was constructed using mtDNA ND3 sequences of nine brachycladiid species, and host phylogeny using cytochrome b sequences of 104 mammalian species. A total of 50 host-parasite links were identified. Distance-based methods supported the hypothesis of a global non-random association of host and parasite phylogenies. Significant individual links (i.e., 24 out of 50) were those related to Campula oblonga, Nasitrema delphini, N. globicephalae and Brachycladium atlanticum and their associated taxa from the Delphinoidea. Regarding event-based methods, we explored 54 schemes using different combinations of costs for each potential coevolutionary event. Three coevolutionary scenarios were identified across all schemes and in all cases the number of loss events (87-156) was the most numerous, followed by failure-to-diverge (40), duplication (3-6), host-switching (0-3) and cospeciation (0-2). We developed a framework to interpret the evolution of this host-parasite system and confirmed that failure-to-diverge and colonization with or without subsequent diversification could have been decisive in the establishment of the associations between brachycladiids and marine mammals.

Life-history trade-offs in a generalist digenean from cetaceans: the role of host specificity and environmental factors.

BackgroundAdults and larvae of generalist parasites are exposed to diverse hosts and local environmental conditions throughout their life cycles, thus local adaptation is expected to occur through phenotypic plasticity and/or natural selection. We investigated how the combined effect of cryptic host specificity and local selective pressures could shape reproductive traits of a putative generalist parasite in the oceanic realm.MethodsThe LSU rDNA, ITS2 and the mt-COI of individuals of the digenean Pholeter gastrophilus (Kossack, 1910) Odhner, 1914 (Heterophyidae Leiper, 1909) from oceanic striped dolphins, Stenella coeruleoalba Meyen, and coastal bottlenose dolphins, Tursiops truncatus Montagu, in the western Mediterranean were used to elucidate whether worms were conspecific. Infection parameters were compared between both dolphin species. General Linear Mixed Models were used to analyse the influence of host species on four reproductive traits of P. gastrophilus: body size, maturity stage (non-gravid/gravid), egg size, and number of eggs in utero. AIC values were used to rank competing models, and p-values to assess the effect of specific predictors.ResultsEvidence indicated that worms collected from both dolphin species were conspecific. All worms collected were gravid and infection parameters did not differ between dolphin species. However, body size and egg size of individuals of P. gastrophilus were significantly larger in striped dolphins. The number of eggs in utero did not significantly differ between dolphin species but, for a given body size, worms in bottlenose dolphins harboured more eggs. A trade-off between egg size and egg number was found in worms from both dolphin species, with a higher slope in striped dolphins.ConclusionsApparently, striped dolphin is a more suitable host for P. gastrophilus, but reproductive investment seems to be adapted to the habitat where the life-cycle develops. Worms from striped dolphins likely face the problem of finding intermediate hosts in the oceanic realm and apparently invest into offspring size to enhance the early survival of larvae and the potential to multiply asexually within the first intermediate host. The small-sized worms from bottlenose dolphins would be adapted to reproduce early because of higher adult mortality, generating smaller and numerous eggs in a coastal habitat where chances of transmission are presumably higher.

Long-Distance Travellers: Phylogeography of a Generalist Parasite, Pholeter gastrophilus, from Cetaceans.

We studied the phylogeography and historical demography of the most generalist digenean from cetaceans, Pholeter gastrophilus, exploring the effects of isolation by distance, ecological barriers and hosts’ dispersal ability on the population structure of this parasite. The ITS2 rDNA, and the mitochondrial COI and ND1 from 68 individual parasites were analysed. Worms were collected from seven oceanic and coastal cetacean species from the south western Atlantic (SWA), central eastern Atlantic, north eastern Atlantic (NEA), and Mediterranean Sea. Pholeter gastrophilus was considered a single lineage because reciprocal monophyly was not detected in the ML cladogram of all individuals, and sequence variability was <1% for mtDNA and 0% for ITS2. These results rule out a recent suggestion that P. gastrophilus would actually be a cryptic-species complex. The genetic cohesion of P. gastrophilus could rely on the extensive exploitation of wide-ranging and highly mobile cetaceans, with a putative secondary role, if any, of intermediate hosts. Unique haplotypes were detected in SWA and NEA, and an AMOVA revealed significant population structure associated to the genetic variation in these regions. The Equator possibly acts as a significant geographical barrier for cetacean movements, possibly limiting gene flow between northern and southern populations of P. gastrophilus. A partial Mantel tests revealed that the significant isolation of NEA populations resulted from geographic clustering. Apparently, the limited mobility of cetaceans used by P. gastrophilus as definitive hosts in this region, coupled with oceanographic barriers and a patchy distribution of potential intermediate hosts could contribute to significant ecological isolation of P. gastrophilus in NEA. Rather unexpectedly, no genetic differentiation was found in the Mediterranean samples of this parasite. Historical demographic analyses suggested a recent population expansion of P. gastrophilus in the Atlantic Ocean, perhaps linked to initial association and subsequent spreading in cetaceans.

A new brachycladiid species (Digenea) from Gervais’ beaked whale Mesoplodon europaeus in north-western Atlantic waters

A new species of the digenean family Brachycladiidae Odhner, 1905 is described from the bile ducts of a Gervais’ beaked whale Mesoplodon europaeus Gervais (Ziphiidae) stranded on the North Atlantic coast of Florida. These parasites were assigned to Brachycladium Looss, 1899 and differed from other species of the genus in the relative size of the oral and ventral suckers, the form and size of the eggs and their extremely small body size. A canonical discriminant analysis was used to examine differences between these specimens and the smallest available individuals of B. atlanticum (Abril, Balbuena and Raga, 1991) Gibson, 2005, considered the morphologically closest species. The overall results exhibited significant differences between the two samples and a jack-knife classification showed that 96.2% of the specimens were correctly classified to their group. In view of evidence from morphological data, the specimens from M. europaeus are considered as new to science and are designated as Brachycladium parvulum n. sp.

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, Cuviers 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.