Francisco Javier Aznar

Morphological and molecular characterization of tetraphyllidean merocercoids (Platyhelminthes: Cestoda) of striped dolphins ( Stenella coeruleoalba ) from the Western Mediterranean

Two types of tetraphyllidean merocercoids, Phyllobothrium delphini and Monorygma grimaldii, are well known from most cetaceans world-wide. The role of cetaceans in the life-cycle of these merocercoids is unclear because their specific identity is as yet unknown. The problem is compounded by poor descriptions of both merocercoids. We used light and scanning electron microscopy, and histological techniques to provide a thorough description of merocercoids collected from 11 striped dolphins, Stenella coeruleoalba, from the Spanish Mediterranean. We also described, for the first time, specimens of P. delphini with immature proglottides. Our merocercoids were morphologically similar to those described previously, except in the structure of the apical organ. Intra- and inter-sample variability in the morphology of the apical organ suggested that it degenerates during larval development. A subsample of 16 specimens of P. delphini and M. grimaldii was characterized for the D2 variable region of the large subunit ribosomal RNA gene (LSU) and compared with published tetraphyllidean cestode LSU sequences. P. delphini showed 2 unique signatures that differed from one another by a single base, whereas all sequences of M. grimaldii were identical. This suggests that each type may represent a single species, contrary to previous speculations based on morphological data. All merocercoid specimens formed a clade together with Clistobothrium montaukensis. Based on the low degree of divergence, all specimens of this clade are predicted to be congeneric.

Epidemiological pattern of tattoo skin disease: a potential general health indicator for cetaceans.

The presence of tattoo skin disease (TSD) was examined in 1392 free-ranging and dead odontocetes comprising 17 species from the Americas, Europe, South Africa, New Zealand and Greenland. We investigated whether TSD prevalence varied with sex, age and health status. TSD was encountered in cetaceans from the Pacific and Atlantic Oceans as well as in those from the North, Mediterranean and Tasman Seas. No clear patterns related to geography and host phylogeny were detected, except that prevalence of TSD in juveniles and, in 2 species (dusky dolphin Lagenorhynchus obscurus and Burmeisters porpoise Phocoena spinipinnis), in adults was remarkably high in samples from Peru. Environmental factors and virus properties may be responsible for this finding. Sex did not significantly influence TSD prevalence except in the case of Peruvian P. spinipinnis. Generally, there was a pattern of TSD increase in juveniles compared to calves, attributed to the loss of maternal immunity. Also, in most samples, juveniles seemed to have a higher probability of suffering TSD than adults, presumably because more adults had acquired active immunity following infection. This holo-endemic pattern was inverted in poor health short-beaked common dolphins Delphinus delphis and harbour porpoises Phocoena phocoena from the British Isles, and in Chilean dolphins Cephalorhynchus eutropia from Patagonia, where adults showed a higher TSD prevalence than juveniles. Very large tattoos were seen in some adult odontocetes from the SE Pacific, NE Atlantic and Portugals Sado Estuary, which suggest impaired immune response. The epidemiological pattern of TSD may be an indicator of cetacean population health.

A NEW CYSTIDICOLID NEMATODE FROM MULLUS SURMULETUS (PERCIFORMES: MULLIDAE) FROM THE WESTERN MEDITERRANEAN

Ascarophis valentina n. sp. is described from Mullus surmuletus off the Valencian coast of Spain on the basis of both light and scanning electron microscopy. It can be distinguished from the other members of the genus by the length of the left (long) spicule of the males and by egg morphology. An updated grouping of the species of Ascarophis considered valid is provided with respect to these characters. The new species resembles Ascarophis capelanus, belonging to the group of species possessing eggs with a single polar knob with filaments, but is distinguished by the size of the body, the length of the esophagus (especially in relation to body length), the position of the vulva, and the size of the left spicule. The new species also shows substantial morphological differences compared with the 3 species, Ascarophis mullusi, Ascarophis upenei, and Ascarophis parupenei, previously described from mullid hosts.

Corynosoma cetaceum in the Stomach of Franciscanas, Pontoporia blainvillei (Cetacea): an Exceptional Case of Habitat Selection by an Acanthocephalan

Adult acanthocephalans are typically found in the intestine of vertebrates, where they can readily absorb nutrients. However, Corynosoma cetaceum has been frequently reported in the stomach of cetaceans from the Southern Hemisphere. The ecological significance of this habitat was investigated by examining data on number, sex ratio, maturity status, biomass, and fecundity of C. cetaceum in different parts of the digestive tract of 44 franciscanas Pontoporia blainvillei. Individual C. cetaceum occurred in the pyloric stomach (PS) and, to lesser degrees, in the duodenal ampulla (DA) and the main stomach (MS). Females outnumbered males in all chambers, although the sex ratio was closer to 1:1 in the MS; there also was a higher proportion of nongravid females, with a smaller biomass in the MS than in the PS and the DA. This evidence suggests that cystacanths are released from prey tissues in the MS, where entire prey are reduced to semi-fluid chyme. The 3 chambers harbored gravid females that did not differ significantly in mean biomass or fecundity. The maturity status of females was nearly identical between the PS and the DA. In the MS, the higher proportion of non-gravid females is probably due to the occurrence of newly recruited females to this site. Mean biomass and fecundity of gravid females covaried strongly and positively among chambers within hosts. These results suggest that there are no major differences between the 3 chambers with respect to the suitability for reproduction by C. cetaceum. However, although the MS is the largest chamber, it harbored the smallest number of gravid females. Interestingly, worms were largely restricted to the aboral portion of the MS, a sheltered region where a concentration of chyme, and thus nutrient availability, likely occurs. Linear distribution differences of gravid female C. cetaceum at increasing intensities suggest that reproductive females occupy chambers according to available space. In summary, the stomach should be considered the main habitat for C. cetaceum. The choice of this habitat is puzzling because other Corynosoma species occur in the intestine, and because the stomach of cetaceans is not an absorptive site.

Molecular phylogeny of the families Campulidae and Nasitrematidae (Trematoda) based on mtDNA sequence comparison

Historically, the systematic arrangement of the genera within the family Campulidae, and its relationship with its allied family Nasitrematidae have been rather confused, particularly because only adult morphology has been available to classical taxonomic analysis. In this paper we provide a partial phylogeny of the genera of these families based on mtDNA from five campulid species: Campula oblonga, Zalophotrema atlanticum, Hadwenius tursionis, Oschmarinella rochebruni and Orthosplanchnus fraterculus; and one nasitrematid, Nasitrema globicephalae. Fasciola hepatica and Dicrocoelium dendriticum were used as outgroups. Maximum parsimony and neighbour-joining methods were applied. Both methods produced similar trees where H. tursionis appeared as the basal campulid, with a sequential divergence of Z. atlanticum, N. globicephalae, C. oblonga, O. rochebruni and O. fraterculus. Results suggest that Nasitrematidae as defined should loose its familial status and the current subfamilial division of the family Campulidae is at least partly artificial and should not be maintained.

Helminth Community Structure in Birds of Prey (Accipitriformes and Falconiformes) in Southern Italy

abstract:  We compared helminth communities in 6 species of birds of prey from the Calabria region of southern Italy. In total, 31 helminth taxa, including 17 nematodes, 9 digeneans, 3 acanthocephalans, and 2 cestodes, were found. All helminth species were observed in the gastrointestinal tract, except for 3 spirurid nematodes. Most of the parasite species were detected in at least 2 hosts, but 13 helminth species were found in only 1 host. At the infracommunity level, the overall species richness and Brillouins index of diversity varied by host, with the highest values in a generalist feeder, the Eurasian buzzard (Buteo buteo), and the lowest in a specialist, the western honey buzzard (Pernis apivorus). Species richness was gender dependent only in the sparrow hawk (Accipiter nisus). The helminth communities were characterized by different dominant species, namely, Centrorhynchus spp. (Acanthocephala) in the Eurasian buzzard and common kestrel (Falco tinnunculus), Parastrigea intermedia (Digenea) in the marsh harrier (Circus aeruginosus), Physaloptera alata (Nematoda) in the sparrow hawk, Serratospiculum tendo (Nematoda) in the peregrine falcon (Falco peregrinus), and Strigea falconis (Digenea) in the western honey buzzard. Statistical analyses confirmed a highly significant difference of helminth infracommunity structure among host species. We conclude that in the Calabria region of southern Italy, each of the raptor species studied is distinct in terms of its helminth communities, and more diverse feeding habits of the host correspond with richer helminth communities.

TETRAPHYLLIDEAN PLEROCERCOIDS FROM WESTERN MEDITERRANEAN CETACEANS AND OTHER MARINE MAMMALS AROUND THE WORLD: A COMPREHENSIVE MORPHOLOGICAL ANALYSIS

Tetraphyllidean plerocercoids have occasionally been reported in marine mammals, but they have rarely been described in detail, and the ecological significance of these infections is unclear. We described plerocercoids collected from the mucosa of the terminal colon and rectum, the anal crypts, and the hepatopancreatic ducts of 7 striped dolphins Stenella coeruleoalba, 1 Cuviers beaked whale Ziphius cavirostris, and 3 Rissos dolphins Grampus griseus from the Spanish Mediterranean. We also examined undescribed plerocercoids from 3 cetacean species from the Atlantic and the Pacific. All plerocercoids had a lanceolate body, and a scolex with an apical sucker and 4 sessile monolocular bothridia. The bothridia had free posterior edges and an accessory sucker at their anterior end. Under light microscopy, the bothridia of some Mediterranean specimens looked bilocular without accessory suckers, but a true accessory sucker was observed in histological sections. A principal component analysis revealed 2 stable clusters of specimens along the first principal component regardless of host species. These “large” and “small” morphotypes are thought to represent early migratory stages of Phyllobothrium delphini and Monorygma grimaldii. The similarity in scolex morphology, the observation of plerocercoids buried in intestinal regions close to the sites where M. grimaldii and P. delphini occur, and the coexistence of all larval forms in the same individual hosts would support this hypothesis. Future molecular analysis may confirm it.

Helminth communities of loggerhead turtles (Caretta caretta) from Central and Western Mediterranean Sea: The importance of host's ontogeny

We investigated the factors providing structure to the helminth communities of 182 loggerhead sea turtles, Caretta caretta, collected in 6 localities from Central and Western Mediterranean. Fifteen helminth taxa (10 digeneans, 4 nematodes and 1 acanthocephalan) were identified, of which 12 were specialist to marine turtles; very low numbers of immature individuals of 3 species typical from fish or cetaceans were also found. These observations confirm the hypothesis that phylogenetic factors restrict community composition to helminth species specific to marine turtles. There were significant community dissimilarities between turtles from different localities, the overall pattern being compatible with the hypothesis that parasite communities reflect the ontogenetic shift that juvenile loggerheads undergo from oceanic to neritic habitats. The smallest turtles at the putative oceanic, pelagic-feeding stage harboured only the 2 digenean species that were regionally the most frequent, i.e. Enodiotrema megachondrus and Calycodes anthos; the largest turtles at the putative neritic, bottom-feeding stage harboured 11 helminth taxa, including 3 nematode species that were rare or absent in turtles that fed partially on pelagic prey. Mean species richness per host was low (range: 1.60-1.89) and did not differ between localities. Variance ratio tests indicated independent colonization of each helminth species. Both features are expected in ectothermic and vagrant hosts living in the marine environment.

Anisakid Larvae in the Musculature of the Argentinean Hake, Merluccius hubbsi

We report the infection levels of third-stage anisakid larva in the muscle of the Argentinean hake, Merluccius hubbsi, in relation to fish size and location in the musculature. The musculature of 42 hake was separated into hypaxial (ventral) and epiaxial (dorsal) parts and surveyed for nematode larvae. Two anisakid species were detected: Anisakis sp. (prevalence, 52.4%; mean +/- SD abundance, 1.2 +/- 1.7) and Pseudoterranova sp. (prevalence, 9.5%; mean +/- SD abundance, 0.2 +/- 0.7). Since the fish were gutted after capture, the occurrence of anisakids in the flesh indicates that the worms had migrated into the muscle before capture. The number of Anisakis sp. in muscle was not correlated with fish length or weight. Therefore, fish size cannot be used as a predictor of parasite loads in the muscle. Only one Anisakis sp. and one Pseudoterranova sp. appeared in the epiaxial musculature. The density of Anisakis sp. in the hypaxial muscles was significantly higher than that in the epiaxial ones. This suggests that removal of the hypaxial musculature can reduce the risk of anisakid-induced allergies and gastrointestinal anisakidoses among consumers.

Parasites as fish population tags and pseudoreplication problems: the case of striped red mullet Mullus surmuletus in the Spanish Mediterranean

Studies of parasites as fish population tags often apply a single round of sampling to identify potential stocks or predict harvest localities. However, the lack of replication generates pseudoreplication, implicitly assuming that infection levels are more similar between samples from the same locality than between samples from different localities. We evaluated this assumption in the case of the striped red mullet Mullus surmuletus in three localities of the Spanish Mediterranean separated by c. 300 km. Samples of 25 fish of similar size were collected in each locality in the summer and autumn of two consecutive years. Prevalence and abundance of three long-lived parasite taxa differed significantly among localities, indicating their potential as stock indicators. However, a cluster analysis (for prevalence) and a MANOVA (for abundance) indicated strong inter-sample variability, even within the same locality, with poor spatial segregation among samples. A linear discriminant analysis (LDA) based on the abundance of 17 parasite taxa correctly assigned over 80% of fish to their locality, and 95% bootstrap confidence intervals of percent classified fish per locality were narrow, indicating good and stable predictive power. However, when a LDA based on data from the first year was used to predict the locality of fish from the second year, predictive power dropped drastically (46% of correct allocation). Overall, we interpret that parasite communities of mullets change at a much lower spatial scale than that adopted in this study. This finding strongly suggests the need for proper replication to make reliable inferences about stock structure in fish populations based on parasitological data.