Mass mortality event of round sardinella Sardinella aurita Valenciennes associated with Glugea Thélohan, 1891 microsporidian infection off the southern Italian coast.

Intracellular parasites of the genus Glugea Thélohan, 1891 (Microsporidia) comprise about 34 putative species capable of causing high morbidity and mortality in freshwater and marine teleost fishes. In this study, we report on the first mass mortality event associated with Glugea sp. infecting free-ranging round sardinella Sardinella aurita in the southern Tyrrhenian Sea (Italy). Here, we describe the ultrastructure of mature spores of this microsporidian and characterize it molecularly, as well as report its phylogenetic position. Most of the affected fish showed an irregular swelling of its abdomen. At necropsy, a variable number of xenomas, spherical to ellipsoidal in shape, were found in the peritoneal cavity strongly attached to the viscera of all fish. Histological analysis revealed varying severity of chronic inflammation along with occasional necrosis in visceral organs associated with multiple xenoma proliferation. These pathological findings were considered the main cause of this mass mortality event. Morphologically, the present material was closely related to G. sardinellesis and G. thunni. The phylogenetically closest taxa to the newly SSU rDNA sequence were G. thunni and an erroneusly identified  G. plecoglossi, which were very closely related to each other, also suggesting that all these sequences might belong to the same species.

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.

First record of the cirripede Anelasma squalicola Darwin, 1852 (Pollicipedidae) in the Mediterranean Sea.

Anelasma squalicola Darwin 1852, is a barnacle that occurs on elasmobranchs from the Atlantic, Pacific, Arctic and Indian Ocean. It infects sharks of the families Etmopteridae and Pentanchidae. The barnacle attaches sub-dermally to its hosts, relying on its interstitial fluid for its nourishment. Herein we present the first record of this species in the Mediterranean Sea, from Etmopterus spinax (Linnaeus, 1758) caught during a bottom-trawl survey in the Alboran Sea (western Mediterranean) in 2017. The overall prevalence of infection was very low (0.22%). The calculated HSI index for the parasitized individual was lower compared to overall means of other individuals and could indicate a potential physiological alteration in the host.

Genetic variation of Sparicotyle chrysophrii (Monogenea: Microcotylidae) from the gilthead sea bream Sparus aurata (Teleostei: Sparidae) in the Mediterranean Sea.

The gill monogenean Sparicotyle chrysophrii (Van Beneden & Hesse, 1863) Mamaev, 1984 is a specific and common parasite of wild and cultured gilthead sea bream Sparus aurata Linnaeus, 1758, able to cause disease and mortality in aquaculture systems. Few molecular studies have been carried out on this monogenean, and its population structure and genetic diversity are barely known. This study provides the first contribution to the population genetic variation of S. chrysophrii, based on two molecular markers - the structural ribosomal RNA (rRNA) for the large subunit (28S) and the cytochrome c oxidase subunit I (COI) gene. Samples were collected from the gills of farmed and wild S. aurata from Italy and the Spanish Mediterranean. The analysis included previously published sequences. The 28S rDNA analysis was consistent with previous studies of specimens isolated from S. aurata and confirmed the presence of only one species on the gills of this host in the Mediterranean Sea. The COI sequences analysis suggested that the samples isolated in a previous study from a different host species, wild Boops boops (Linnaeus, 1758) in the Adriatic Sea, may represent a new undescribed sister species of S. chrysophrii. The low nucleotide diversity of S. chrysophrii isolated only from S. aurata versus the high haplotype diversity revealed small differences between haplotypes. The haplotypes shared between wild and farmed hosts from Spain provided the first molecular evidence of the possible transfer of S. chrysophrii between wild and farmed populations of S. aurata. The mtDNA COI analysis did not show a clear genetic structure, probably the result of several factors including coevolution, wild and farmed host interactions, and host population structure in space and time.

Infection process, viability and establishment of Anisakis simplex s.l. L3 in farmed fish; A histopathological study in gilthead seabream.

Anisakis spp. (Nematoda, Anisakidae) are parasites known by their economic and health impacts, as their L3 larval stages infect a variety of fish species, many of them commercial species, sometimes causing zoonotic episodes due to consumption of raw or undercooked fish. The aim of this study is to determine the infection process and the potential impact of A. simplex s.l. L3 on gilthead seabream (Sparus aurata L.), one of the most important fish species in Mediterranean aquaculture, by periodic histological monitoring of the infection process. For this, fish were experimentally infected with A. simplex s.l. L3 and periodically analysed for L3 larvae, collecting samples at different time points (hours post ingestion, hpi): 3, 6, 12, 18, 24, 36, 48, 72, 96, 120, 144, 168 and 192, up to 6 months post infection (mpi). All samples were observed under a stereomicroscope and later fixed for histological examination. A. simplex s.l. L3 were only found on the visceral surface and mesenteric tissue, but never free or encapsulated in muscle. Chronological events were found to occur faster than those reported in previous studies. They were first observed 6 hpi in the coelomic cavity, being present up to 48 hpi. While the earliest evidence of fibrocytes surrounding A. simplex s.l. L3 larvae were observed at 18 hpi, complete spiral encapsulation occurred by 72 hpi. Alive parasites were observed up to 6 mpi. Although the infection of gilthead seabream by Anisakis spp. larvae is feasible, it seems unlikely, especially in aquaculture given the hygienically controlled feeding systems. In the event of infection, the transmission would be unlikely due to the poor condition in which specimens of Anisakis spp. are found. Furthermore, since no larvae were detected in the fish's muscle, human infection seems improbable.

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.

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.

Acanthocephalans from Marine Fishes from Patagonia, Argentina.

In this study, 542 individual fish from 20 species from the Patagonian continental shelf of Argentina were examined for acanthocephalans. A total of 1,547 acanthocephalans belonging to 5 species were collected from 18 species of fish. Adult forms were represented by 2 species: Aspersentis johni ( Baylis, 1929 ) (Heteracanthocephalidae) from longtail southern cod, Patagonotothen ramsayi (Regan) (new host record), and Breizacanthus aznari Hernández-Orts, Alama-Bermejo, Crespo, García, Raga and Montero, 2012 (Arhythmacanthidae) from raneya, Raneya brasiliensis (Kaup). Immature worms of B. aznari were also collected from the intestine of pink cusk-eel, Genypterus blacodes (Forster) (new host record). Cystacanths of 3 species of Corynosoma Lühe, 1904 (Polymorphidae) were found encapsulated in the mesenteries of fish. Corynosoma australe Johnston, 1937 was the most abundant acanthocephalan in our study, infecting 18 species of fish and accounting for >89.9% of all specimens collected. A cystacanth of Corynosoma bullosum (Linstow, 1892) was found in "castañeta", Nemadactylus bergi (Norman) (new host record), and cystacanths of Corynosoma cetaceum Johnston and Best, 1942 were collected from red searobin, Prionotus nudigula Ginsburg, and flounders Paralichthys isosceles Jordan (new host record) and Xystreurys rasile (Jordan). The Patagonian shelf of Argentina represents a new locality record for A. johni and C. bullosum. This survey is a starting point for understanding the diversity of marine acanthocephalans in Patagonian waters.

Transmission of Corynosoma australe (Acanthocephala: Polymorphidae) from fishes to South American sea lions Otaria flavescens in Patagonia, Argentina.

Acanthocephalans display a two-host life cycle that involves arthropods as intermediate hosts and vertebrates as definitive hosts. Some species also use paratenic hosts to bridge the trophic gap between both obligatory hosts. However, the relative role of these paratenic hosts in the transmission to definitive hosts has seldom been assessed quantitatively. We report on infection patterns of cystacanths of Corynosoma australe Johnston, 1937 in 20 common teleost species and the Argentine shortfin squid Illex argentinus (Castellanos) from the Patagonian shelf of Argentina. We also explore the role of different fish species in the transmission of C. australe to the most important definitive host in the area, i.e. the South American sea lion Otaria flavescens Shaw. Cystacanths of C. australe were found in all host species except Heliconus lahillei Norman, Merluccius hubbsi Marini and I. argentinus. In eight fish species, the prevalence of C. australe was > 50% and mean intensity > 4, i.e. Acanthistius patachonicus (Jenyns), Nemadactylus bergi (Norman), Paralichthys isosceles Jordan, Percophis brasiliensis Quoy & Gaimard, Prionotus nudigula Ginsburg, Scomber colias Gmelin, Raneya brasiliensis (Kaup) and Xystreurys rasile (Jordan). Two surveys on the trophic ecology of South American sea lions in the study area consistently found a generalist diet dominated by M. hubbsi, and data on the frequency of occurrence and number of other fish and cephalopod species in stomach contents strongly suggest that only R. brasiliensis may play a prominent role in the transmission of C. australe. This result raises interesting questions on the costs of paratenicity.

Parasite communities of the white seabream Diplodus sargus sargus in the marine protected area of Medes Islands, north-west Mediterranean Sea.

Marine protected areas are considered a useful tool to preserve and recover the biodiversity of ecosystems. It is suggested that fisheries not only affect populations of target and bycatch species but also their parasite communities. Parasites can indicate fishery effects on host species and also on the whole local community, but the effects of fisheries and protection measures on parasite communities are relatively unknown. This study analyses parasite communities of the white seabream Diplodus sargus sargus in order to assess potential effects exerted by protection measures within and by fisheries outside a reserve in the western Mediterranean Sea. This small scale analysis offered the opportunity to study different degrees of fishery effects on parasite infracommunities, without considering climatic effects as an additional factor. Parasite infracommunities of fishes from the no-take zone (NTZ) differed in their composition and structure compared with areas completely or partially open to fisheries. The detected spatial differences in the infracommunities derived from generalist parasites and varied slightly between transmission strategies. Monoxenous parasites were richer and more diverse in both fished areas, but more abundant in the no-take, whereas richness and abundance of heteroxenous parasites were higher for the NTZ. In addition to host body size as one factor explaining these spatial variations, differences within parasite infracommunities between the areas may also be linked to increased host densities and habitat quality since the implementation of the NTZ and its protection measures.

Diagnostic accuracy of the light microscope method to detect the eggs of Cardicola spp. in the gill filaments of the bluefin tuna.

Trematode blood flukes of the genus Cardicola are potentially lethal in bluefin tuna cultures. The present study proposed a new method to detect aporocotylid eggs in tuna gills. Aporocotylid eggs were detected by analysing a pair of gill filaments of five transversal areas of the eight holobranches of one hundred Atlantic bluefin tuna and observed with glycerol and a stereomicroscope with an oblique brightfield. Data were gathered according to holobranches, transversal areas and their combination. Eggs were uniformly distributed among the holobranches, but they had the highest prevalence in the second and fifth transversal areas, which is controversial with respect to previous studies of egg distribution. An abbreviated method called the T-two test, which had the highest sensitivity (96.8%), is proposed for the detection of Cardicola spp. infections instead of the analysis all the holobranches. The T-two test limits the time and cost of the egg parasite screening analysis. The analyses of ten samples could be sufficient to detect the presence of parasites in farmed bluefin tuna; fish from the wild are expected to be less infected and more samples (45) would therefore be necessary.

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.

Ecology of the Atlantic black skipjack Euthynnus alletteratus (Osteichthyes: Scombridae) in the western Mediterranean Sea inferred by parasitological analysis.

Between 2008 and 2011, the head of 150 Euthynnus alletteratus (Osteichthyes: Scombridae) caught inshore off the southeastern Iberian coast (western Mediterranean Sea) were examined for parasites. Two monogeneans, four didymozoid trematodes and four copepods were found. Parasite abundance showed a positive relationship with the annual sea surface temperature, except for Pseudocycnus appendiculatus, but negative with the sea depth (Capsala manteri, Neonematobothrium cf. kawakawa and Caligus bonito). Prevalences and mean abundances differed significantly among sampling areas, except for C. manteri, Oesophagocystis sp. 2 and Ceratocolax euthynni, and sampling years (Melanocystis cf. kawakawa, N.cf. kawakawa, P. appendiculatus and Unicolax collateralis). Results indicate that the parasite abundances of E. alletteratus in the western Mediterranean Sea depend mainly on regional environmental variables, which can show interannual variations. The presence of pelagic parasites, i.e. didymozoids and P. appendiculatus, could indicate that E. alletteratus migrates between inshore and offshore pelagic domains. The different parasite faunas reported in E. alletteratus populations from the western Atlantic Ocean and the Mediterranean Sea appear to point out the geographical host isolation. These results suggest that E. alletteratus inhabiting the western Mediterranean Sea performs inshore-offshore small-scale migrations, and not transoceanic migrations between the western Atlantic Ocean and Mediterranean Sea.

Morphological and attachment changes of Lamellodiscus theroni (Monogenea: Diplectanidae) during its post-larval development on fish.

Species of the genus Lamellodiscus Johnston et Tiegs, 1922 (Monogenea: Diplectanidae) are characterised by a complex haptor bearing many different attachment elements: two pairs of main hooks joined by medial bars, 14 peripheral marginal hooks and one or two lamellodiscs, formed by several overlapping sclerotised plates (lamellae). These haptoral structures appear gradually during parasite development and, therefore, attachment strategies vary with developmental stage. The main aim of this work was to study the developmental changes of Lamellodiscus theroni Amine, Euzet et Kechemir-Issad, 2007 under experimental conditions, with special attention to the gradual variations in attachment strategies and the pathological implications. Throughout the gradual development of the sclerotised structures, six developmental phases were distinguished in L. theroni: phase I, with only 14 peripheral marginal hooks; phase II, with main hooks (ventral and dorsal) formed; phase III, with ventral bar formed; phase IV, with dorsal bars formed; phase V, with dorsal and ventral lamellodiscs formed; and phase VI, adult stage with male copulatory organ formed. During development, parasites attach to different parts of the first and secondary gill lamellae and the mode of attachment changes from unspecific stage, i.e. based on piercing any flat gill tissue in the early stages, through an intermediate stage when ventral and dorsal main hooks are completely functional and parasites become restricted to the interlamellar space, and finally to the definitive adult attachment stage when lamellodiscs are fully developed. The timing of key events in the development of L. theroni was used to establish adequate intervals for anthelmintic drug administration.

Raphidascaris (Raphidascaris) macrouri n. sp. (Nematoda: Anisakidae) from two deep-sea macrourid fishes in the Western Mediterranean: Morphological and molecular characterisations.

A new nematode species, Raphidascaris (Raphidascaris) macrouri n. sp. (Anisakidae), is described from male and female specimens found in the intestine, and occasionally in stomach and pyloric caeca, of two deep-water macrourid fishes (Gadiformes) off Barcelona, Mediterranean Sea: Nezumia aequalis (Günther) (type-host) and Trachyrincus scabrus (Rafinesque). Based on light and scanning electron microscopy examination, the new species shows similar morphological features as the other four valid species of the subgenus Raphidascaris Railliet & Henry, 1915, but it differs from Raphidascaris (Raphidascaris) acus (Bloch, 1779), Raphidascaris (Raphidascaris) lutjani Olsen, 1952 and Raphidascaris (Raphidascaris) mediterraneus Lèbre & Petter, 1983 in the high number of precloacal papillae (23-32) and from Raphidascaris (Raphidascaris) gigi Fujita, 1928 in the length of the spicules. Moreover, Raphidascaris (Raphidascaris) macrouri n. sp. exhibits a high variability on the number and distribution of caudal papillae, which was not recorded in the other four mentioned species. This is the first species of this subgenus reported from the family Macrouridae. Sequences of ITS1-5.8S-ITS2 region are analysed and compared with closely related nematode species. Molecular analysis confirmed the uniformity of the R. (R.) macrouri n. sp. between hosts.

Morphological characterisation and identification of four species of Cardicola Short, 1953 (Trematoda: Aporocotylidae) infecting the Atlantic bluefin tuna Thunnus thynnus (L.) in the Mediterranean Sea.

Blood flukes of the genus Cardicola Short, 1953 are considered the most potentially pathogenic parasites in bluefin tuna cultures. Morphological study and genetic analyses of the ribosomal internal transcribed spacer ITS-2 and the mitochondrial cytochrome c oxidase 1 (cox1) gene fragments revealed the occurrence of four aporocotylid species (C. forsteri Cribb, Daintith & Munday, 2000, C. orientalis Ogawa, Tanaka, Sugihara & Takami, 2010, C. opisthorchis Ogawa, Ishimaru, Shirakashi, Takami & Grabner, 2011 and Cardicola sp.) in 421 Thunnus thynnus (L.) from the Western Mediterranean (274 fished from the wild and 147 from sea-cages). Cardicola opisthorchis was the most abundant species, with higher prevalence in the cage-reared fish than in those fished in the wild (21 vs 6%, p < 0.05). Adults of three species were recovered: C. forsteri from both gills and heart, C. opisthorchis from heart and C. orientalis from gills. The secondary gill lamellae were profusely infected by eggs of C. orientalis. A fourth species was found in four tunas, based on the molecular analyses of eggs apparently indistinguishable in size and shape from the eggs of C. orientalis. The findings provided evidence that infections with Cardicola spp. differed in relation to locality, host origin (wild vs cage-reared) and site of infection. It is necessary to estimate the possible different pathogenic effects of each species of Cardicola in order to take appropriate control measures.

Risk assessment for parasites in cultures of Diplodus puntazzo (Sparidae) in the Western Mediterranean: prospects of cross infection with Sparus aurata.

The sharpsnout seabream Diplodus puntazzo is of interest in Mediterranean fish farming. Disease is an important problem because parasites can spread quickly in culture conditions and fish often develop high parasite burdens. Here we assess the risk that documented parasites pose to the sustainability of D. puntazzo farming. This study specifically considers metazoan and protist parasites recorded from wild and farmed D. puntazzo in scientific literature. Risk assessment studies involve the identification, characterization and qualitative quantification of the risk in question (parasitoses in this case) and the probability of establishment. We considered the parasite species which may be difficult to manage as a priority for research into potential management strategies. Those parasites which could be transmitted from cultures of gilthead seabream (Sparus aurata) were also included in this study. Four groups of parasites represented a risk to D. puntazzo farming, ranging from moderate to high: Ciliophora, Myxozoa, Monogenea and Copepoda. Three parasite species were considered high risk to D. puntazzo cultures: Amyloodinium sp., Cryptocaryon sp. and Enteromyxum leei. These species were responsible for high mortalities in cultures of these and other fish species. In addition Sparicotyle chrysophrii, Caligus ligusticus and Gnathia vorax entail a moderate risk to D. puntazzo Mediterranean farms. No important episodes have been related to caligids and isopods in Mediterranean sparids, nevertheless they should be properly managed to prevent future problems.

Description, microhabitat selection and infection patterns of sealworm larvae (Pseudoterranova decipiens species complex, nematoda: ascaridoidea) in fishes from Patagonia, Argentina.

BACKGROUND: Third-stage larvae of the Pseudoterranova decipiens species complex (also known as sealworms) have been reported in at least 40 marine fish species belonging to 21 families and 10 orders along the South American coast. Sealworms are a cause for concern because they can infect humans who consume raw or undercooked fish. However, despite their economic and zoonotic importance, morphological and molecular characterization of species of Pseudoterranova in South America is still scarce. METHODS: A total of 542 individual fish from 20 species from the Patagonian coast of Argentina were examined for sealworms. The body cavity, the muscles, internal organs, and the mesenteries were examined to detect nematodes. Sealworm larvae were removed from their capsules and fixed in 70% ethanol. For molecular identification, partial fragments of the mitochondrial cytochrome c oxidase subunit 1 gene (cox1) were amplified for 10 isolates from 4 fish species. Morphological and morphometric data of sealworms were also obtained. RESULTS: A total of 635 larvae were collected from 12 fish species. The most infected fish was Prionotus nudigula, followed by Percophis brasiliensis, Acanthistius patachonicus, Paralichthys isosceles, and Pseudopercis semifasciata. Sequences obtained for the cox1 of sealworms from A. patachonicus, P. isosceles, P. brasiliensis and P. nudigula formed a reciprocally monophyletic lineage with published sequences of adult specimens of Pseudoterranova cattani from the South American sea lion Otaria flavescens, and distinct from the remaining 5 species of Pseudoterranova. A morphological description, including drawings and scanning electron microscopy photomicrographs of these larvae is provided. Sealworms collected from Argentinean fishes did not differ in their diagnostic traits from the previously described larvae of P. cattani. However a discriminant analysis suggests that specimens from P. nudigula were significantly larger than those from other fishes. Most of the sealworms were collected encapsulated from the muscles and, to a lesser degree, from the mesenteries and the liver. CONCLUSIONS: We provided the first molecular identification, morphological description and microhabitat characterization of sealworm larvae from the Argentinean Patagonian coast. We also reported the infection levels of sealworms on 20 fish species in order to elucidate the life cycle of these nematodes in this area.

Seasonal population dynamics of Zeuxapta seriolae (Monogenea: Heteraxinidae) parasitising Seriola dumerili (Carangidae) in the Western Mediterranean.

We examined the seasonal and yearly population dynamics of the monogenean pathogen Zeuxapta seriolae on juvenile fish from wild populations of Seriola dumerili. The study is based on bimonthly monitoring between April, 2005 and April, 2007 off Majorca, and newly obtained monogenean population data for juvenile fish from three additional localities in the Western Mediterranean (off Alicante, Corsica and Sardinia). We documented the highest intensities and abundances of Z. seriolae, with mean abundance values similar to or higher than those reported in the single case of wild fish mortalities reported to date. There was a recurrent pattern of seasonal change in infection with Z. seriolae in the populations of S. dumerili off Majorca, with substantially higher parasite loads during the warm season (April to June). Mean parasite abundance was significantly correlated with seawater temperature and associated with higher proportions of juvenile worms in the parasite populations, thus suggesting increased transmission rates at higher temperatures. There was a significant negative association between abundance of Z. seriolae and fish length. Comparisons with the samples of younger and older fish off Majorca indicated that whereas infection parameters gradually increased in the first year of juvenile fish life, larger/older fish (>43 cm; 1+) were much lightly infected than the smaller/younger (<30 cm; 0+) juvenile fish examined in the same season. The observed increases in abundance during the warm weather months were invariably associated with sharp increases in monogenean aggregation levels and this was in contrast with the markedly low levels for both parameters during the cold season months. These data, coupled with the strong negative correlation between the levels of aggregation of Z. seriolae and mean fish total length, indicate that heavily infected individuals are being rapidly removed (i.e. within 2-3 months) from the host population thus reducing the heterogeneity of parasite distribution as fish grow. We discuss parasite-induced host mortality and other mechanisms that may account for observed recurrent patterns in monogenean abundance and spatial aggregation and review the data available on the spread of Z. seriolae infections in the Mediterranean.

Redescriptions of two frequently recorded but poorly known hemiurid digeneans, Lecithochirium musculus (Looss, 1907) (Lecithochiriinae) and Ectenurus lepidus Looss, 1907 (Dinurinae), based on material from the western Mediterranean.

Two frequently reported but poorly known hemiurid digeneans, Lecithochirium musculus (Looss, 1907) (Lecithochiriinae), from the stomach of Trachinus draco and Citharus linguatula, and Ectenurus lepidus Looss, 1907 (Dinurinae), from the stomach of Spicara maena, are redescribed based on material from off the Barcelona coast of the western Mediterranean. The two species are commented upon, and Lecithochirium israelense Fischthal, 1980 is considered a synonym of L. musculus. Records of the two species in the Mediterranean Basin and North East Atlantic region are summarised.