Determinants of diversity and composition of the tapeworm fauna of blue sharks, Prionace glauca: a geographical and host-specificity analysis.

Blue sharks, Prionace glauca, are cosmopolitan, extremely vagile sharks and the species among elasmobranchs for which most surveys containing tapeworm community data are available worldwide. In this study we report on the tapeworm fauna of three samples of blue sharks (n = 37) from two new regions (one sample from Galicia, north-east Atlantic, and two from Valencia, western Mediterranean), and compared it with previous studies, assessing the relative role of the ecological and evolutionary factors in structuring local tapeworm assemblages. Nine cestode taxa were identified, of which four included adult specimens, that is, Platybothrium auriculatum, Prosobothrium armigerum, Anthobothrium caseyi and Molicola horridus. The abundance of these species, and Brillouin's diversity index, differed significantly among samples without a clear geographical signal. A comparison with six previous surveys revealed that tapeworm assemblages were composed of the same 'core' taxa, with mean species richness typically ranging from two to four species. Global records of adult tapeworms in blue sharks included: 15 taxa identified at species level, of which only eight (generalist trypanorhynchs) were shared with other sympatric host species; five mostly with other carcharhinids; and three with large lamnid sharks sharing the blue sharks' habitat. The composition of tapeworm communities of blue sharks is thus highly constrained by strong host specificity, with composition and abundance varying across localities depending on idiosyncratic environmental conditions.

Epibiotic fauna of the Antarctic minke whale as a reliable indicator of seasonal movements.

Antarctic minke whales, Balaenoptera bonaerensis, breed in tropical and temperate waters of the Southern Hemisphere in winter and feed in Antarctic grounds in the austral summer. These seasonal migrations could be less defined than those of other whale species, but the evidence is scanty. We quantitatively describe the epibiotic fauna of Antarctic minke whales and explore its potential to trace migrations. Seven species were found on 125 out of 333 examined Antarctic minke whales captured during the last Antarctic NEWREP-A expedition in the Southern Ocean: the amphipod Balaenocyamus balaenopterae (prevalence = 22.2%), the copepod Pennella balaenoptera (0.6%); three coronulid, obligate barnacles, Xenobalanus globicipitis (11.1%), Coronula reginae (8.7%), C. diadema (0.9%); and two lepadid, facultative barnacles, Conchoderma auritum (9.0%) and C. virgatum (0.3%). Species with prevalence > 8% exhibited a modest increase in their probability of occurrence with whale body length. Data indicated positive associations between coronulid barnacles and no apparent recruitment in Antarctic waters. All specimens of X. globicipitis were dead, showing progressive degradation throughout the sampling period, and a geographic analysis indicated a marked drop of occurrence where the minimum sea surface temperature is < 12 °C. Thus, field detection -with non-lethal methodologies, such as drones- of coronulid barnacles, especially X. globicipitis, on whales in the Southern Ocean could evince seasonal migration. Future investigations on geographical distribution, growth rate, and degradation (for X. globicipitis) could also assist in timing whales' migration.

Bottlenose dolphins (Tursiops truncatus) aggressive behavior towards other cetacean species in the western Mediterranean.

Aggressive behavior of bottlenose dolphins (Tursiops truncatus) towards conspecifics is widely described, but they have also often been reported attacking and killing harbour porpoises (Phocoena phocoena) around the world. However, very few reports exist of aggressive interactions between bottlenose dolphins and other cetacean species. Here, we provide the first evidence that bottlenose dolphins in the western Mediterranean exhibit aggressive behavior towards both striped dolphins (Stenella coeruleoalba) and Risso's dolphins (Grampus griseus). Necropsies and visual examination of stranded striped (14) and Risso's (2) dolphins showed numerous lesions (external rake marks and different bone fractures or internal organ damage by blunt trauma). Indicatively, these lessons matched the inter-tooth distance and features of bottlenose dolphins. In all instances, these traumatic interactions were presumed to be the leading cause of the death. We discuss how habitat changes, dietary shifts, and/or human colonization of marine areas may be promoting these interactions.

Recent increase of ulcerative lesions caused by Anisakis spp. in cetaceans from the north-east Atlantic.

Species of Anisakis typically infect the stomach of cetaceans worldwide, often causing ulcerative lesions that may compromise the host's health. These nematodes also cause anisakiasis or allergic reactions in humans. To assess the risks of this emerging zoonosis, data on long-term changes in Anisakis infections in cetaceans are necessary. Here, we compare the prevalence and severity of ulcerative lesions caused by Anisakis spp. in five cetacean species stranded along the north-west Spanish coast in 2017-2018 with published data from 1991-1996. Open ulcers were found in 32/43 short-beaked common dolphins, Delphinus delphis; 3/5 striped dolphins, Stenella coeruleoalba; 1/7 bottlenose dolphins, Tursiops truncatus; and 1/3 harbour porpoises, Phocoena phocoena meridionalis; a single individual of long-finned pilot whale, Globicephala melas, was found uninfected. In common dolphins, the mean abundance of open ulcers per host was 1.1 (95% confidence interval: 0.8-1.3), with a maximum diameter (mean ± standard deviation) of 25.4 ± 16.9 mm. Stomachs with scars or extensive fibrosis putatively associated with Anisakis were detected in 14 and five animals, respectively. A molecular analysis based on the mitochondrial cytochrome c oxidase II gene using 18 worms from three cetacean species revealed single or mixed infections of Anisakis simplex sensu stricto and Anisakis pegreffii. Compared with the period 1991-1996, we found a strong increase of prevalence, abundance and extension of ulcerative lesions in most cetacean species. Anisakis populations could have increased in the study area over the last decades, although we cannot rule out that a higher environmental stress has also boosted the pathological effects of these parasites.

Two decades of monitoring in marine debris ingestion in loggerhead sea turtle, Caretta caretta, from the western Mediterranean.

Anthropogenic marine debris is one of the major worldwide threats to marine ecosystems. The EU Marine Strategy Framework Directive (MSFD) has established a protocol for data collection on marine debris from the gut contents of the loggerhead sea turtle (Caretta caretta), and for determining assessment values of plastics for Good Environmental Status (GES). GES values are calculated as percent turtles having more than average plastic weight per turtle. In the present study, we quantify marine debris ingestion in 155 loggerhead sea turtles collected in the period 1995-2016 in waters of western Mediterranean (North-east Spain). The study aims (1) to update and standardize debris ingestion data available from this area, (2) to analyse this issue over two decades using Zero-altered (hurdle) models and (3) to provide new data to compare the only GES value available (off Italian waters). The composition of marine debris (occurrence and amounts of different categories) was similar to that found in other studies for the western Mediterranean and their amounts seem not to be an important threat to turtle survival in the region. Model results suggest that, in the study area, (a) period of stranding or capture, (b) turtle size and (c) latitude are significant predictors of anthropogenic debris ingestion (occurrence and amount) in turtles. The GES value for late juvenile turtles (CCL>40 cm) has decreased in the last ten years in the study area, and this is very similar to that obtained in Italian waters. We also provide a GES value for early juvenile turtles (CCL≤40 cm) for the first time. Recommendations arising from this study include ensuring use of (1) the standardized protocol proposed by the MSFD for assessing marine debris ingestion by loggerhead sea turtles and (2) the ecology of the turtles (neritic vs oceanic), rather than their size, to obtain GES values.

Parasitic outbreak of the copepod Balaenophilus manatorum in neonate loggerhead sea turtles (Caretta caretta) from a head-starting program.

BACKGROUND: Diseases associated to external parasitosis are scarcely reported in sea turtles. During the last decades several organism have been documented as a part of normal epibiont community connected to sea turtles. The copepod Balaenophilus manatorum has been cited as a part of epibiont fauna with some concern about its parasitic capacity. This study serves three purposes, i.e. (i) it sheds light on the type of life style that B. manatorum has developed with its hosts, particularly turtles; (ii) it makes a cautionary note of the potential health risks associated with B. manatorum in sea turtles under captivity conditions and in the wild, and (iii) it provides data on effective treatments against B. manatorum. RESULTS: We report for the first time a massive infestation of the copepod B. manatorum and subsequent acute mortality in a group of loggerhead sea turtle hatchlings. Four-month-old turtles from a head-starting program started exhibiting excitatory and fin rubbing behavior preceding an acute onset of lethargy, skin ulceration and death in some animals. All the individuals (n = 57) were affected by severe copepod load and presented different degrees of external macroscopic skin lesions. The ventral area of front flippers, axillar and pericloacal skin were mostly affected, and were the main parasite distribution regions. Copepods were also detected on plastron and carapace sutures. The gut contents of B. manatorum reacted positively for cytokeratin, indicating consumption of turtle skin. Severe ulcerative necrotic dermatitis and large amount of bacteria presence were the major histopathological findings. CONCLUSIONS: Individual fresh water immersion for 10 min and lufenuron administration (0.1 ppm) to the water system every 2 weeks proved effective for removing turtle parasites and to control re-infestation, respectively. The results from our study clearly indicated that B. manatorum individuals consume turtle skin. The pathological effects of this agent and the potential implications in sea turtle conservation and management are discussed.

Nyctiphanes couchii as intermediate host for Rhadinorhynchus sp. (Acanthocephala, Echinorhynchidae) from NW Iberian Peninsula waters.

In the mesozooplanktonic community of the coastal upwelling system of the Ría de Vigo (NW Spain), the euphausiid Nyctiphanes couchii has been identified for the first time in temperate waters of the NE Atlantic as the intermediate host for cystacanths of Rhadinorhynchus sp. Parasites were identified using morphological characters described in 20 cystacanths. The hooks of the proboscis were arranged in 14 rows of 26 hooks each, while the hooks of the basal circle were only slightly erected and were longer than remaining spines. A maximum-likelihood estimation (ML) tree inferred from the 18S rRNA data set of Palaeacantocephala revealed that our specimens belong to a highly supported clade with Rhadinorhynchus sp., Pararhadinorhynchus sp. and Transvena annulospinosa. Nonetheless, our morphological and phylogenetic analyses suggested that the status of Rhadinorhynchus pristis should be re-examined. The prevalences of parasites were 0.0019% and 0.0001% for frontal and coastal summer communities, and 0.0068% and 0.0008% for coastal and oceanic autumn communities, respectively. The presence of these cystacanths in different mesozooplankton communities throughout the study suggests that the recruitment of parasites may be affected by the oceanography.

Lousy mums: patterns of vertical transmission of an amphibious louse.

In this study, we document patterns of vertical transmission of the amphibious louse Antarctophthirus microchir (Echinophthiriidae) in pups of South American sea lion, Otaria flavescens, from Patagonia. Vertical transmission is fundamental for the long-term stability of A. microchir populations because only pups stay long enough (1 month) on land for the louse to reproduce. A total of 72 pups ≤7 days old from a single rookery were captured and examined for lice. Infection parameters and population structure of A. microchir did not differ among pups collected at the beginning, middle, and end of the reproductive season, suggesting that patterns of early vertical transmission are not affected by the increase of rookery size during this period. Over 60% of 1-day-old pups were infected with A. microchir, and recruitment increased in pups up to 3 days old and then leveled off. In 1-day-old pups, significantly more adults than nymphs were found, but the pattern was reversed in older pups. The number of first-stage nymphs was significantly smaller than that of second- and third-stage nymphs, as it was the number of males vs. females, particularly in 1-day-old pups. Three non-exclusive hypotheses could account for these patterns, i.e., recruitment merely reflects the population structure of A. microchir is cows; the relative ability of lice to pass from cows onto pups increases in advanced instars; and/or natural selection favors transmission of adults, especially females, because they accrue greater fitness. The importance of latter hypothesis should not be underestimated in a species with a tight reproductive schedule.

Parasite assemblages in the Western whip snake Hierophis viridiflavus carbonarius (Colubridae) from southern Italy.

Parasite assemblages of the Western whip snake Hierophis viridiflavus carbonarius were investigated from the Calabria region in southern Italy. A total of 14 parasite taxa including 6 nematodes, 3 acanthocephalans, 2 cestodes, 2 digeneans and a single pentastomid was identified. Within the study area, H. v. carbonarius serves as the final host for seven species of helminths, of which only four (Hexametra quadricornis, Kalicephalus viperae, Paracapillaria sonsinoi and Renifer aniarum) can be considered as snake specialists, while one (Oswaldocruzia filiformis) is shared with other reptiles and amphibians, and two (Paradistomum mutabile and Rhabdias fuscovenosa) with lizards. A large proportion of larval forms of six helminth taxa (about 95% of all helminths collected) was found, for which H. v. carbonarius serves as an intermediate and/or paratenic host; however, adult stages of helminths were prevalent in snakes with snout-to-vent length greater than 70 cm. Our results suggest that ontogenetic and ecological factors should exert a strong influence upon the helminth assemblage of Western whip snakes. We concluded that H. v. carbonarius plays an important role in southern Italy as an intermediate/paratenic host for species of helminths infecting vertebrate groups which may include this snake species within their feeding chain. Eleven taxa, including three potential agents of zoonosis, were added to the poorly known parasite fauna of this host.

Intestinal helminth fauna of the South American sea lion Otaria flavescens and fur seal Arctocephalus australis from northern Patagonia, Argentina.

We report on the intestinal helminth fauna of 56 South American sea lions, Otaria flavescens, and 5 South American fur seals, Arctocephalus australis, from northern Patagonia, Argentina. A total of 97,325 helminth specimens were collected from sea lions. Gravid individuals were represented by 6 species of parasites: 1 digenean (Ascocotyle (Ascocotyle) patagoniensis), 1 cestode (Diphyllobothrium spp.), 3 nematodes (Uncinaria hamiltoni, Contracaecum ogmorhini s.s., Pseudoterranova cattani) and 1 acanthocephalan (Corynosoma australe). In addition, third-stage larvae of 2 nematodes (Contracaecum sp. and Anisakis sp. type I) and 3 juvenile acanthocephalans (Andracantha sp., Profilicollis chasmagnathi and Corynosoma cetaceum) were also collected. Andracantha sp., C. ogmorhini s.s. and P. chasmagnathi represent new host records. A total of 1516 helminth specimens were collected from fur seals. Gravid individuals were represented by three species of parasites, namely, Diphyllobothrium spp., C. ogmorhini s.s. and C. australe. In addition, larvae of Contracaecum sp. and P. cattani, juveniles of C. cetaceum and immature cestodes (Tetrabothriidae gen. sp.) were also collected. Corynosoma australe was the most prevalent and abundant parasite in both hosts, accounting for >90% of all specimens. Sea lions and furs seals from northern Patagonia harbour the intestinal helminth communities that could be predicted for otariids, i.e. the combination of species of the genera Corynosoma, Diphyllobothrium, Pseudoterranova, Contracaecum and, in pups, Uncinaria. Additionally, both species of otariid are apparently unsuitable hosts (i.e. non-hosts) for as many as five parasite taxa. The inclusion or exclusion of these species affects estimation of species richness at both component community (11 versus 6 species in sea lions; 7 versus 3 species in fur seals) and infracommunity (mean: 3.1 versus 2.6 in sea lions; 2.2 versus 1.7 species) levels. Information about the reproductive status of helminth species is often lacking in parasitological surveys on otariids and other marine vertebrates, but it is of significance to improve precision in parascript studies or ecological meta-analyses.

Nyctiphanes couchii as intermediate host for the acanthocephalan Bolbosoma balaenae in temperate waters of the NE Atlantic.

Cystacanths of the acanthocephalan Bolbosoma balaenae (Gmelin, 1790) were found encapsulated in the cephalothorax of the euphausiid Nyctiphanes couchii (Bell, 1853) from temperate waters in the NE Atlantic Ocean. Euphausiids were caught in locations outside the Ría de Vigo in Galicia, NW Spain, and prevalence of infection was up to 0.1%. The parasite was identified by morphological characters. Cystacanths were 8.09 ± 2.25 mm total length (mean ± SD) and had proboscises that consisted of 22 to 24 longitudinal rows of hooks, each of which had 8 or 9 hooks per row including 2 or 3 rootless ones in the proboscis base and 1 field of small hooks in the prebulbar part. Phylogenetic analyses of 18S rDNA and cytocrome c oxidase subunit I revealed a close relationship with other taxa of the family Polymorphidae (Meyer, 1931). The results extend northwards ot the known distribution of B. balaenae. Taxonomic affiliation of parasites and trophic ecology in the sampling area suggest that N. couchii is the intermediate host for B. balenae, and we suggest that the whales Balaenoptera physalus (Linnaeus, 1758) and B. acutorostrata (Lacepède, 1804) are its definitive hosts. This life cycle is probably completed with or without paratenic hosts.

Life begins when the sea lion is ashore: microhabitat use by a louse living on a diving mammal host.

Among Anoplura, the family Echinophthiriidae includes species that infest pinnipeds and otters. Previous evidence obtained from pinnipeds infested by echinophthiriids, specifically from seals, indicates that flippers are the preferred infestation sites, while lice from fur seals select areas in the pelage. We studied habitat selection of Antarctophthirus microchir on South American sea lion pups (Otaria flavescens Shaw, 1800) from Patagonia, Argentina, during the austral summer of 2009. We found a clear pattern of habitat selection: eggs are laid on the dorsal surface; nymphs 1 hatch there and then migrate to the belly, where they develop into adults and copulate; and then ovigerous females return to the dorsal surface. On the one hand, nymphs 1 are characterised by their low locomotory ability; therefore, the fact that they migrate as soon as they hatch suggests a clear pressure leading to microhabitat restriction. On the other hand, the described pattern of microhabitat selection seems to respond to the physiological requirements of each stage, which vary according to the physiological process considered, e.g. oviposition, morphogenesis, hatching and development. Accordingly, it appears that A. microchir would prefer the host's ventral area for development and copulation and the dorsal area for oviposition. However, the causes of this pattern are not clear, and many factors could be involved. Considering that sea lion pups periodically soak at high tides, and that prolonged immersion and very high humidity are known to be lethal for lice eggs, selecting the dorsal area would be advantageous for oviposition because it dries much faster. Furthermore, because humidity should be retained for longer periods on the ventral surface of the pup, wetter conditions on the sea lion would prevent desiccation of the nymphs in the very arid environment where O. flavescens breeds.

Assessing host-parasite specificity through coprological analysis: a case study with species of Corynosoma (Acanthocephala: Polymorphidae) from marine mammals.

In this paper we report an investigation of the utility of coprological analysis as an alternative technique to study parasite specificity whenever host sampling is problematic; acanthocephalans from marine mammals were used as a model. A total of 252 scats from the South American sea lion, Otaria flavescens, and rectal faeces from 43 franciscanas, Pontoporia blainvillei, from Buenos Aires Province, were examined for acanthocephalans. Specimens of two species, i.e. Corynosoma australe and C. cetaceum, were collected from both host species. In sea lions, 78 out of 145 (37.9%) females of C. australe were gravid and the sex ratio was strongly female-biased. However, none of the 168 females of C. cetaceum collected was gravid and the sex ratio was not female-biased. Conversely, in franciscanas, 14 out of 17 (82.4%) females of C. cetaceum were gravid, but none of 139 females of C. australe was, and the sex ratio of C. cetaceum, but not that of C. australe, was female-biased. In putative non-hosts, the size of worms was similar to that from specimens collected from prey. Results suggest that both acanthocephalans contact sea lions and franciscanas regularly. However, C. australe and C. cetaceum cannot apparently reproduce, nor even grow, in franciscanas and sea lions, respectively. Coprological analysis may represent a useful supplementary method to investigate parasite specificity, particularly when host carcasses are difficult to obtain.

Helminth community structure of the Mediterranean gull (Ichthyaetus melanocephalus) in Southern Italy.

The helminth community of the Mediterranean gull (Ichthyaetus melanocephalus) from the Calabria region in southern Italy was examined. Ten species of helminths, including 2 cestodes (Alcataenia larina and Tetrabothrius cylindraceus), 4 digeneans ( Aporchis massiliensis , Brachylaima fuscatum, Cardiocephaloides longicollis, and Ornithobilharzia canaliculata), and 4 nematodes (Eucoleus contortus, Capillaria sp., Cosmocephalus obvelatus, and Paracuaria adunca), were found. The Mediterranean gull represents new host records for A. larina, A. massiliensis, B. fuscatum, and E. contortus; all species, except C. longicollis and O. canaliculata, constitute new locality records. The gastrointestinal helminth community of I. melanocephalus consisted of 9 species but, at the infracommunity level, species richness, diversity, and total helminth abundance are among the lowest reported from any gull species. Two cestode species accounted for 81% of all helminth specimens found.

Host specificity of Oschmarinella rochebruni and Brachycladium atlanticum (Digenea: Brachycladiidae) in five cetacean species from western Mediterranean waters.

We investigated patterns of specificity of liver flukes (fam. Brachycladiidae) in a community of cetaceans from the western Mediterranean. The liver and pancreas of 103 striped dolphins, Stenella coeruleoalba, 18 Risso's dolphins, Grampus griseus, 14 bottlenose dolphins, Tursiops truncatus, 8 common dolphins, Delphinus delphis, and 5 long-finned pilot whales, Globicephala melas, were analysed for brachycladiid species. Two species were found: Oschmarinella rochebruni in striped dolphins (prevalence (P): 61.2%; mean intensity (MI) (95% CI): 34.2 (25.7-45.6)), and Brachycladium atlanticum in striped dolphins (P: 39.8%; MI: 7.1 (4.8-13.1)) and a single individual of common dolphin (P: 12.5%; intensity: 19), which represents a new host record. A molecular analysis using the internal transcribed spacer 2 (ITS2) region of the rDNA gene confirmed that specimens of B. atlanticum were conspecific regardless of host species. Available dietary data suggest that Risso's dolphins, bottlenose dolphins and long-finned pilot whales would contact rarely, if at all, the infective stages of O. rochebruni and B. atlanticum. Neither the prevalence nor the mean abundance of B. atlanticum differed significantly between striped and common dolphins, but a principal component analysis using seven morphometric variables indicated that specimens collected from the common dolphin were stunted. These worms also had fewer eggs compared with specimens typically found in striped dolphins, although the size of the eggs was similar in both host species. Dwarfism and low fecundity have typically been found in helminths infecting unusual host species, and might reflect the lower compatibility of B. atlanticum for common dolphins. In summary, both O. rochebruni and B. atlanticum appear to exhibit a narrow specificity for striped dolphins in the western Mediterranean.

Population dynamics of Antarctophthirus microchir (Anoplura: Echinophthiriidae) in pups from South American sea lion, Otaria flavescens, in Northern Patagonia.

We analysed population dynamics of the louse Antarctophthirus microchir in pups of the South American sea lion, Otaria flavescens, at the Punta León rookery (Argentina) over a period of 2 years. A total of 136 pups were aged and marked at the beginning of the lactation period ashore, then sampled for lice at different times within 30 days. Sampling was restricted to the chest and belly, two sites where lice were especially abundant. This concentration on ventral areas might protect lice from thermal stress in the austral summer. Infestation patterns in pups 3 days old suggested that the potential for transmission increased from first nymphs to adults. Population trends of each instar with pup age, based on standardised values of abundance, were conserved between years, reflecting the basic dynamics of recruitment and reproduction. However, trends based on log-transformed abundances varied significantly between years; apparently, environmental conditions affected growth of lice populations differently each year. Stage-based deterministic models for population growth of A. microchir suggested generation times from 18 to 23 days. Accordingly, only 2 lice generations might be produced before pups start going to the sea. Shortening the cycle to accommodate a third generation might be risky, whereas a 2-generation cycle might at least result in larger females producing higher numbers of viable offspring.

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.

Insight into the role of cetaceans in the life cycle of the tetraphyllideans (Platyhelminthes: Cestoda).

Four types of tetraphyllidean larvae infect cetaceans worldwide: two plerocercoids differing in size, 'small' (SP) and 'large' (LP), and two merocercoids referred to as Phyllobothrium delphini and Monorygma grimaldii. The latter merocercoid larvae parasitize marine mammals exclusively and exhibit a specialised cystic structure. Adult stages are unknown for any of the larvae and thus the role of cetaceans in the life cycle of these species has been a long-standing problem. The SP and LP forms are thought to be earlier stages of P. delphini and M. grimaldii that are presumed to infect large pelagic sharks that feed on cetaceans. A molecular analysis of the D2 variable region of the large subunit ribosomal DNA gene based on several individuals of each larval type collected from three Mediterranean species of cetaceans showed consistent and unique molecular signatures for each type regardless of host species or site of infection. The degree of divergence suggested that LP, P. delphini and M. grimaldii larvae may represent separate species, whereas SP may be conspecific with M. grimaldii. In all host species, individuals of SP accumulated in the gut areas in which the lymphoid tissue was especially developed. We suggest therefore that these larvae use the lymphatic system to migrate to the abdominal peritoneum and mesenteries where they develop into forms recognizable as M. grimaldii. The plerocercoid stage of P. delphini remains unknown. In a partial phylogenetic tree of the Tetraphyllidea, all larvae formed a clade that included a representative of the genus Clistobothrium, some species of which parasitize sharks such as the great white which is known to feed on cetaceans. A bibliographic examination of tetraphyllidean infections in marine mammals indicated that these larvae are acquired mostly offshore. In summary, the evidence suggests that cetaceans play a significant role in the life cycle of these larvae. In addition, it seems clear that cetaceans act as natural intermediate hosts for P. delphini and M. grimaldii, as within these hosts they undergo development from the plerocercoid stage to the merocercoid stage. Because tetraphyllidean species use fish, cephalopods and other marine invertebrates as intermediate hosts, the inclusion of cetaceans in the life cycle would have facilitated their transmission to apex predators such as the large, lamnid sharks. The biological significance of infections of LP in cetaceans is unclear, but infections do not seem to be accidental as such larvae show high prevalence and abundance as well as a high degree of site specificity, particularly in the anal crypts and bile ducts.

Distribution of Pholeter gastrophilus (Digenea) within the stomach of four odontocete species: the role of the diet and digestive physiology of hosts.

We compared the distribution of the digenean Pholeter gastrophilus in the stomach of 27 harbour porpoises, Phocoena phocoena, 27 striped dolphins, Stenella coeruleoalba, 18 bottlenose dolphins, Tursiops truncatus, and 100 long-finned pilot whales, Globicephala melas. The stomach of these species is composed of 4 chambers of different size, structure and function. In all species, P. gastrophilus was largely restricted to the glandular region of the stomach, but the parasite tended to favour the fundic chamber in bottlenose dolphins and harbour porpoises, the pyloric chamber in pilot whales, and none in striped dolphins. However, predictability at infrapopulation level was generally low, suggesting a weak preference of P. gastrophilus for any of the chambers. Three hypotheses were tested to investigate a common cause for the distribution of P. gastrophilus in all host species, namely, colonization of chambers was (1) sequential, (2) dependent on chamber size, or (3) dependent on the passage time of food through the whole stomach. The latter hypothesis was indirectly tested by assuming, based on previous evidence from other vertebrates, that the greater the size of the stomach and/or the energy content of prey, the greater the delay of food passage. We found no compelling evidence that chamber colonization was sequential, or related to chamber size in any species. However, the distribution of P. gastrophilus was significantly more anteriad when the host species had larger stomachs and, particularly, when hosts fed on prey with higher caloric content. Accordingly, the stomach distribution of P. gastrophilus at this scale seems to be passively driven by features of the diet and digestive physiology of each host species. This study provides a general framework to formulate null hypotheses in future studies on microhabitat choice by parasites.

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.