Antibacterial Activities and Life Cycle Stages of Asparagopsis armata: Implications of the Metabolome and Microbiome.

The red alga Asparagopsis armata is a species with a haplodiplophasic life cycle alternating between morphologically distinct stages. The species is known for its various biological activities linked to the production of halogenated compounds, which are described as having several roles for the algae such as the control of epiphytic bacterial communities. Several studies have reported differences in targeted halogenated compounds (using gas chromatography-mass spectrometry analysis (GC-MS)) and antibacterial activities between the tetrasporophyte and the gametophyte stages. To enlarge this picture, we analysed the metabolome (using liquid chromatography-mass spectrometry (LC-MS)), the antibacterial activity and the bacterial communities associated with several stages of the life cycle of A. armata: gametophytes, tetrasporophytes and female gametophytes with developed cystocarps. Our results revealed that the relative abundance of several halogenated molecules including dibromoacetic acid and some more halogenated molecules fluctuated depending on the different stages of the algae. The antibacterial activity of the tetrasporophyte extract was significantly higher than that of the extracts of the other two stages. Several highly halogenated compounds, which discriminate algal stages, were identified as candidate molecules responsible for the observed variation in antibacterial activity. The tetrasporophyte also harboured a significantly higher specific bacterial diversity, which is associated with a different bacterial community composition than the other two stages. This study provides elements that could help in understanding the processes that take place throughout the life cycle of A. armata with different potential energy investments between the development of reproductive elements, the production of halogenated molecules and the dynamics of bacterial communities.

Anguilla sp. diseases diagnoses and treatments: The ideal methods at the crossroads of conservation and aquaculture purposes.

Anguilla anguilla, A. japonica and A. rostrata are the most fished and consumed eel species. However, these species are Critically Endangered, Endangered and Endangered, respectively. A combination of factors is thought to be responsible for their decline including fisheries, climate change, habitat destruction, barriers to migration, pollution and pathogens. Among them, viruses, bacteria and parasites are causing weakening of wild eels and serious economic losses for fishermen and eel farmers. Early detection of pathogens is essential to provide appropriate responses both for conservation reasons and to limit economic losses. Classic diagnosis approaches are time consuming and invasive and usual treatments, for example, antipathogenic substances are becoming obsolete because of pathogen resistance and environmental impact problems. The need for early and non-invasive diagnostic methods as well as effective and environmentally friendly treatments has increased. Vaccine development and diet supplementation have known a growing interest since their use could allow prevention of diseases. In this review, we summarize the main pathogens-viruses, bacteria and parasites-of the three northern temperate eel species, the methods used to detect these pathogens and the different treatments used. We discussed and highlighted the need for non-invasive, rapid and efficient detection methods, as well as effective and environmentally friendly treatments for both conservation and aquaculture purposes.

Molecular detection of the swim bladder parasite Anguillicola crassus (Nematoda) in fecal samples of the endangered European eel Anguilla anguilla.

The European eel Anguilla anguilla is listed as critically endangered by the IUCN. Among many threats, the introduced parasitic nematode Anguillicola crassus is suspected to alter the eels' swim bladder and jeopardize their reproductive oceanic migration. To date, gaining knowledge about the distribution and prevalence of A. crassus requires individual sacrifice (over 50,000 eels were sacrificed for epidemiology studies since 2010). This paper describes a non-lethal molecular protocol for identifying prevalence of A. crassus in A. anguilla, based on searching for A. crassus DNA in the feces of eels. Tests using three DNA microsatellite markers specific to the nematode showed that molecular detection provided similar results to visual examination of the swim bladder in up to 80% of the cases, and allowed for comparison of prevalence among sites. Easy to implement, this non-lethal protocol for detecting A. crassus could be valuable for management plans of this endangered species.

Distribution of anguillid leptocephali and possible spawning areas in the South Pacific Ocean.

Seven South Pacific anguillid eel species live from New Guinea to French Polynesia, but their spawning areas and life histories are mostly unknown despite previous sampling surveys. A July-October 2016 research cruise was conducted to study the spawning areas and times, and larval distributions of South Pacific anguillid eels, which included a short 155°E station-line northeast of New Guinea and five long transects (5-25°S, 160°E-140°W) crossing the South Equatorial (SEC) and other currents. This survey collected nearly 4000 anguilliform leptocephali at 179 stations using an Isaacs-Kidd Midwater Trawl accompanied by 104 CTD casts. Based on mor-phometric observations and DNA sequencing, 74 anguillid leptocephali were collected, which in the southern areas included 29 larvae of six species: Anguilla bicolor pacifica, A. marmorata, A. australis, A. reinhardtii, A. megastoma, and A. obscura (all anguillid species of the region were caught except A. dieffenbachii). Small A. australis (9.0-16.8 mm) and A. reinhardtii (12.4, 12.5 mm) leptocephali were collected south of the Solomon Islands, other A. australis (10.8-12.0 mm) larvae were caught northwest of Fiji along with an A. obscura (20.0 mm) larva, and an A. marmorata (7.8 mm) larva was collected near Samoa. Considering collection sites, larval ages from otolith analysis, and westward SEC drift, multiple spawning locations occurred from south of the Solomon Islands and the Fiji area (16-20 days old larvae) to near Samoa (19 days old larva) during June and July in areas where high-salinity Subtropical Underwater (STUW, ~150 m depth) and the warm, low-salinity surface Fresh Pool were present. Five long hydrographic sections showed the strong Fresh Pool in the west and the STUW formation area in the east.

Lepidapedon sereti n. sp. (Digenea: Lepidapedidae) in Coelorinchus sereti (Gadiformes: Macrouridae) from deep waters off Vanuatu.

Lepidapedon sereti n. sp. is described from the macrourid Coelorinchus sereti from the deep water off Vanuatu. It is placed in the Elongatum group and Elongatum subgroup. It differs from the other species described in this subgroup by the distinctly dorsally subterminal excretory pore. It also differs from other species in combinations of size, excretory vesicle length, proportions of forebody, post-testicular region and other metric features. This constitutes the first record of a Lepidapedon (sensu stricto) from the Central Western Pacific Ocean.

Proctophantastes nettastomatis (Digenea: Zoogonidae) from Vanuatu deep-sea fish: new morphological features, allometric growth, and phenotypic plasticity aspects.

The present paper deals with Proctophantastes nettastomatis (Digenea: Zoogonidae; Lepidophyllinae) found in the intestine of three species of deep-sea fish, Dicrolene longimana (Ophidiidae, Ophidiiformes), Bathyuroconger sp. (Congridae, Anguilliformes), and Venefica tentaculata (Nettastomatidae, Anguilliformes). The fish were collected near the islands of Espiritu Santo, Erromango, and Epi, respectively, in the archipelago of Vanuatu (Southern Pacific Ocean) at depths ranging from 561 to 990 m. Morphological and histological analyses showed that the Vanuatu specimens differ from Proctophantastes abyssorum, Proctophantastes gillissi, Proctophantastes glandulosum, Proctophantastes infundibulum, and Proctophantastes brayi but are close to P. nettastomatis discovered in Suruga Bay, Japan. P. nettastomatis is redescribed based both on the observations of our specimens and of the Japanese holotype and paratype. The morphological variability of the species is described. Morphometric data allowed the identification of positive allometric growth for the hindbody, negative allometric growth for the ventral sucker, and a growth phenotypic plasticity between Ophidiiformes and Anguilliformes definitive hosts.

Assessment of the quality of European silver eels and tentative approach to trace the origin of contaminants - A European overview.

The European eel is critically endangered. Although the quality of silver eels is essential for their reproduction, little is known about the effects of multiple contaminants on the spawning migration and the European eel management plan does not take this into account. To address this knowledge gap, we sampled 482 silver eels from 12 catchments across Europe and developed methods to assess three aspects of eel quality: muscular lipid content (N = 169 eels), infection with Anguillicola crassus (N = 482), and contamination by persistent organic pollutants (POPs, N = 169) and trace elements (TEs, N = 75). We developed a standardized eel quality risks index (EQR) using these aspects for the subsample of 75 female eels. Among 169 eels, 33% seem to have enough muscular lipids content to reach the Sargasso Sea to reproduce. Among 482 silver eels, 93% were infected by A. crassus at least once during their lifetime. All contaminants were above the limit of quantification, except the 1,2-bis(2,4,6-tribromophenoxy)ethane (BTBPE), Ag and V. The contamination by POPs was heterogeneous between catchments while TEs were relatively homogeneous, suggesting a multi-scale adaptation of management plans. The EQR revealed that eels from Warwickshire were most impacted by brominated flame-retardants and agricultural contaminants, those from Scheldt were most impacted by agricultural and construction activities, PCBs, coal burning, and land use, while Frémur eels were best characterized by lower lipid contents and high parasitic and BTBPE levels. There was a positive correlation between EQR and a human footprint index highlighting the capacity of silver eels for biomonitoring human activities and the potential impact on the suitability of the aquatic environment for eel population health. EQR therefore represents a step forward in the standardization and mapping of eel quality risks, which will help identify priorities and strategies for restocking freshwater ecosystems.

Proctophantastes brayi, n. sp. (Digenea: Zoogonidae) parasite of the deep-sea fish Polymixia Lowe, 1838 from Vanuatu.

Proctophantastes brayi n. sp. (Digenea: Zoogonidae; Lepidophyllinae) has been found in the intestine of two species of deep-sea fish Polymixia (silver eye fish) near the island of Erromango in Vanuatu at a depth ranging from 720 to 830 m. Specimen whole mounts, histological and scanning electron microscopy preparations showed that P. brayi differs from the five known species of the genus Proctophantastes (P. abyssorum, P. gillissi, P. glandulosum, P. infundibulum and P. nettastomatis) by the following morphological characters: (i) a slit in the anterior part of the oral sucker, (ii) Laurer's canal is absent, (iii) a more extended periatrial gland than the ones in the other species of Proctophantastes, consisting of divided masses of cells and that form a conspicuous multilobated structure which does not have a membrane-bounded sac, (iv) the distal part of the metraterm has vesicle-like processes which we refer to as metratermal sacs, in addition to atrial sacs, (v) a long extension of the glandular cells surrounding the saccular bladder which extends posteriorly to the excretory pore.

First evidence of European eels exiting the Mediterranean Sea during their spawning migration.

The migration route and the spawning site of the European eel Anguilla anguilla are still uncertain. It has been suggested that the Mediterranean eel stock does not contribute to spawning because there is no evidence of eels leaving the Mediterranean Sea. To test this hypothesis, we equipped eight female silver eels from the south of France with pop-up satellite tags during escapement from coastal waters. Once in deeper water, the eels quickly established diel vertical migration (DVM) between the upper and lower mesopelagic zone. Five tagged eels were taken by predators within the Mediterranean, but two eels reached the Atlantic Ocean after six months and at distances greater than 2000 km from release. These eels ceased their DVM while they negotiated the Gibraltar Strait, and remained in deep water until they reached the Atlantic Ocean, when they recommenced DVM. Our results are the first to show that eels from Mediterranean can cross the Strait of Gibraltar and continue their migration into the Atlantic Ocean. This finding suggests that Mediterranean countries, as for other EU states, have an important role to play in contributing to conservation efforts for the recovery of the European eel stock.

Links between parasitism, energy reserves and fecundity of European anchovy, Engraulis encrasicolus, in the northwestern Mediterranean Sea.

The European anchovy, Engraulis encrasicolus L. 1758, is one of the most sought-after target species in the northwestern Mediterranean Sea. However, this stock currently consists of small individuals, and landings are reported to have decreased considerably. The main purpose of this study was to assess, for the first time, the interrelationships between size, fecundity, energy reserves and parasitism in female anchovies, in order to analyse the potential implications for the health of northwestern Mediterranean anchovy stocks arising from the current shortage of large individuals. Results revealed that smaller individuals show lower fecundity, lower lipid content and a higher intensity of certain parasites. As it is known that smaller individuals now predominate in the population, the relationships found in this study indicate that the health of anchovies from the northwestern Mediterranean is currently impaired.

Bages-Sigean and Canet-St Nazaire lagoons (France): physico-chemical characteristics and contaminant concentrations (Cu, Cd, PCBs and PBDEs) as environmental quality of water and sediment.

Environmental characteristics in water and sediments of two contrasted coastal Mediterranean lagoons, Bages-Sigean and Canet-St Nazaire, were measured over a three season survey. The urban pollution (treatment plant discharges) is very important in Canet-St Nazaire lagoon reflecting untreated sewages, while in Bages-Sigean, the northern part appears more impacted due to larger anthropogenic inputs. Dissolved Cd concentrations are on the whole similar in both lagoons, whereas Cu concentrations are by far higher in lagoon Canet-St Nazaire. Cu concentrations appear to be highly dependent on dissolved organic carbon whereas salinity seems to control Cd variations. Concerning the sediments, the confined northern part of lagoon Bages-Sigean shows organic carbon and total nitrogen enrichment whereas lipid concentrations are much higher in the Canet-St Nazaire lagoon. Cu complexation seems to be strongly related to organic matter as evidenced by the two significant positive relationships, on one hand between Cu and organic carbon, and on the other hand, between Cu and lipids. On the contrary, Cd concentrations appear to be mainly controlled by carbonates. PCBs and PBDEs were detected only in sediments and show relatively low concentrations compared to similar lagoon environments. Regarding the sediment quality guidelines, Cd, Cu and PCBs in both lagoons did not exceed any Probable Effect Concentration (PEC).

Influence of introduced vs. native parasites on the body condition of migrant silver eels.

Because parasitism is among the reasons invoked to explain the collapse of Anguilla anguilla, we evaluated the parasitic constraint on body condition (BC) of migrant silver eels as a proxy of fitness with inter-site comparisons. Metazoan parasites were studied in 149 silver eels from five sites (northern Europe). In total, 89% were infected by 13 species including Myxozoa, Monogenea, Cestoda, Nematoda, and Acanthocephala. Anguillicoloides crassus was most common (56%), then Acanthocephalus clavula (30%), and Pseudodactylogyrus sp. (17%). BC, calculated for 58 females, was negatively correlated by abundance of the introduced Pseudodactylogyrus sp. but not by other parasite taxa. Nevertheless, the introduced A. crassus was considered as a severe pathogen based on previous data, whereas the native A. clavula was supposed to have limited impact. Parasite component communities and BC were different between sites. Silver eels from Stockholm Archipelago (Sweden) were the least parasitized (40% vs. 90-95% for other sites) with no parasites on the gills. Burrishoole (Ireland) differed by the absence of A. crassus and high prevalence of A. clavula (84%) but without consequences on BC. Gudenaa (Denmark), Corrib (Ireland), and Frémur (France) were close due to high prevalence of A. crassus (89-93%). Gudenaa and Corrib were the most similar because Pseudodactylogyrus sp. was also highly prevalent (respectively 71% and 60%) whereas absent in Frémur. Our results suggest that the fitness loss induced by the introduced parasites could affect the spawning success of migrant silver eels from Gudenaa and Corrib, and to a lesser extent from Frémur, but probably not those from Stockholm Archipelago and Burrishoole.

Complete genomic sequence and taxonomic position of eel virus European X (EVEX), a rhabdovirus of European eel.

Eel virus European X (EVEX) was first isolated from diseased European eel Anguilla anguilla in Japan at the end of seventies. The virus was tentatively classified into the Rhabdoviridae family on the basis of morphology and serological cross reactivity. This family of viruses is organized into six genera and currently comprises approximately 200 members, many of which are still unassigned because of the lack of molecular data. This work presents the morphological, biochemical and genetic characterizations of EVEX, and proposes a taxonomic classification for this virus. We provide its complete genome sequence, plus a comprehensive sequence comparison between isolates from different geographical origins. The genome encodes the five classical structural proteins plus an overlapping open reading frame in the phosphoprotein gene, coding for a putative C protein. Phylogenic relationship with other rhabdoviruses indicates that EVEX is most closely related to the Vesiculovirus genus and shares the highest identity with trout rhabdovirus 903/87.

Dynamic expression of immune response genes in the sea bass, Dicentrarchus labrax, experimentally infected with the monogenean Diplectanum aequans.

The sea bass, Dicentrarchus labrax, is one of the most extensively farmed marine fishes in the Mediterranean. Under the high-density condition common in aquaculture, the monogenean gill parasite Diplectanum aequans can cause significant economic losses. This study used real-time quantitative PCR to investigate the dynamic expression of immune response genes in sea bass infected with Diplectanum aequans. The target genes, interleukin-1 (IL-1beta, transforming growth factor (TGF-beta and T-cell receptor (TCR-beta), were studied in the gills and spleen of the sea bass from the first day of infection until thirty days post- infection. Our results showed that there was an increase in IL-1beta gene expression in the spleen and gills and in TGF-beta gene expression in the gills of infected fish. These results show that parasitic infection induced a local inflammatory reaction and that reaction was restricted to the site of infection. Finally, the absence of relationship between TCR-beta expression and the parasitic infection suggests that the adaptive immune system is not involved in the response against this parasite.