Live Freshwater Parasite, Salmincola californiensis (Copepoda: Lernaeopodidae), on the Gills of an Ocean-Migrating Steelhead Trout (Oncorhynchus mykiss) and Discussion on the Origin and Survival of the Parasite at Sea.

Salmincola californiensis is a parasitic copepod of freshwater salmonids in the North Pacific rim countries. Sixteen adult females of the species were found alive on the gills of an ocean-age 4, maturing steelhead trout, Oncorhynchus mykiss, caught in offshore waters (50°30'N, 179°30'W) of the North Pacific Ocean in July 1997. This is the first evidence of live individuals of S. californiensis from ocean-migrating salmonids. When found, copepods were attached to the distal ends of gill filaments, and their bodies were observed to be slowly moving in Petri dishes with seawater. Ocean-migrating steelhead trout comprise individuals originating from western Kamchatka (Russia) and western North America. Based on the date and catch location of the infected fish, it is inferred that it originated from western North America, where it acquired S. californiensis infection in fresh water. As this fish spent about 4 years in the ocean, the copepods likely survived the same period at sea. However, if the fish was a kelt, the survival period of the copepods in the ocean may be shorter than four years. To confirm identification of the copepods, adult females of S. californiensis are briefly described using the specimens collected from the fish.

Parasites in kelp-forest food webs increase food-chain length, complexity, and specialization, but reduce connectance.

We explored whether parasites are important in kelp forests by examining their effects on a high-quality, high-resolution kelp-forest food web. After controlling for generic effects of network size, parasites affected kelp-forest food web structure in some ways consistent with other systems. Parasites increased the trophic span of the web, increasing top predator vulnerability and the longest chain length. Unique links associated with parasites, such as concomitant predation (consumption of parasites along with their hosts by predators) increased the frequency of network motifs involving mutual consumption and decreased niche contiguity of free-living species. However, parasites also affected kelp-forest food web structure in ways not seen in other systems. Kelp-forest parasites are richer and more specialized than other systems. As a result, parasites reduced diet generality and decreased connectance in the kelp forest. Although mutual consumption motifs increased in frequency, this motif type was still a small fraction of all possible motifs, so their increase in frequency was not enough to compensate for the decrease in connectance caused by adding many specialist parasite species.

Independent Losses of the Hypoxia-Inducible Factor (HIF) Pathway within Crustacea.

Metazoans respond to hypoxic stress via the hypoxia-inducible factor (HIF) pathway, a mechanism thought to be extremely conserved due to its importance in monitoring cellular oxygen levels and regulating responses to hypoxia. However, recent work revealed that key members of the HIF pathway have been lost in specific lineages (a tardigrade and a copepod), suggesting that this pathway is not as widespread in animals as previously assumed. Using genomic and transcriptomic data from 70 different species across 12 major crustacean groups, we assessed the degree to which the gene HIFα, the master regulator of the HIF pathway, was conserved. Mining of protein domains, followed by phylogenetic analyses of gene families, uncovered group-level losses of HIFα, including one across three orders within Cirripedia, and in three orders within Copepoda. For these groups, additional assessment showed losses of HIF repression machinery (EGLN and VHL). These results suggest the existence of alternative mechanisms for cellular response to low oxygen and highlight these taxa as models useful for probing these evolutionary outcomes.

Integrative taxonomy of a new species of Therodamas (Ergasilidae) infecting the Amazonian freshwater fish Leporinus fasciatus (Anostomidae).

Crustaceans of the subclass Copepoda are an important component of the invertebrate aquatic fauna. They occur in all aquatic environments and include some representatives that are free-living organisms and others that have a parasitic lifestyle. The genus Therodamas comprises marine and freshwater copepods whose females are parasites of fish in their adult phase, with only seven species described so far. During a field survey of fish parasites in the Jari River, a large tributary of the Amazon River system, in Brazil, we found a new species of the genus Therodamas infecting Leporinus fasciatus. Therodamas longicollum n. sp. is the second strictly freshwater species known. Phylogenetic analysis showed that the new species is grouped in the family Ergasilidae, and divergence estimates showed that T. longicollum n. sp. diverged from its ancestor at around 66.34 Ma, in the late Upper Cretaceous. Therodamas longicollum n. sp. differs from its congeneric in that it does not have lobes and/or expansion of the anterior neck region. Besides describing a new Therodamas species, thereby increasing the diversity of the genus to eight species, this study points out the existence of a lineage of these copepods that has adapted to the freshwater environment of the Amazon. This study also corroborates the genus Therodamas as part of the family Ergasilidae.

Resting egg banks can facilitate recovery of zooplankton communities after short exposure to glyphosate.

We assessed the short-term viability and recovery of zooplankton communities after exposure to glyphosate (active ingredient-a.i.). We conducted a hatching experiment in two steps: Step 1-natural lake sediments containing resting egg banks were placed into individual trays and exposed to a solution medium of glyphosate at three different treatments (LD = Values below the detection limits, LD < 0.05, 0.44, and 0.89 mg a.i./L) for 14 days; and Step 2-we replaced the exposure solution of glyphosate with distilled freshwater, keeping them all trays under freshwater conditions for another 14 day. The results from Step 1 showed significant effects of glyphosate on the emergence patterns of resting eggs, with a reduction in hatching of rotifers, mainly at concentrations of 0.44 and 0.89 mg a.i./L. On the other hand, the results from Step 2 showed an increase in the emergence of viable eggs for rotifers after restoration of freshwater conditions in all treatments; there was no effect for total zooplankton and microcrustaceans. These findings suggest that (i) glyphosate may, effectively, impair zooplankton hatching from resting egg banks; (ii) the magnitude of the negative effects depends on the the zooplanktonic group considered; and (iii) the restoration of freshwater conditions may, in some way, allow the recovery of the zooplankton community from viable egg banks. Our results can be useful in predicting the influence of glyphosate on the distribution patterns of freshwater zooplankton, which can represent vital information for environmental managers.

Unravelling tidal effect on zooplankton community structure in a tropical estuary.

The variability in zooplankton density and species composition in response to tidal oscillations were investigated in the lesser saline upper reaches (E1) and higher saline lower reaches (E2) of the Mahanadi Estuary, located at the mouth of the third largest river in Indian Peninsula. This is the first of its kind high frequency observation over the tidal cycle ranging from highest high tide to lowest low tide in this estuary revealing dynamic variability of zooplankton assemblages. Zooplankton abundance was higher during high tide in comparison to low tide, irrespective of salinity regimes. On the diurnal scale, it was higher at night in comparison to the day at both E1 and E2. The higher abundance of zooplankton groups such as Copepoda, Cladocera, and planktonic larvae during the night at E1 as well as E2 indicating an upward migration. Many of the zooplankton taxa (e.g., Pseudodiaptomus serricaudatus, Pseudodiaptomus sp., Acartia danae, Acrocalanus longicornis, Oithona sp., Corycaeus andrewsi) migrated towards E1 due to tidal effect during high tide and maintained their position even during low tide. In contrast, the prevalence of limnetic taxa (e.g., Brachionus rubens, Polyarthra vulgaris, Bosminopsis deitersi, Moina micrura, Heliodiaptomus sp.) at E2 during low tide indicated a predominant riverine source. The tidal variability of Brachyura (zoea and megalopa) revealed different emergence times that indicated dispersal of zoeas to the adjacent Bay of Bengal and the return of magalopa to the Mahanadi Estuary. Species diversity index was higher during high tide, and prominent at E2. At E1, marine, fresh, marine-brackish, and marine-brackish-fresh zooplankton species dominated during high tide, while brackish-fresh taxa dominated during low tide. Differently, E2 was enriched with marine, marine-brackish, and marine-brackish-fresh taxa during high tide, whereas fresh, brackish-fresh, and marine-fresh dominated during low tide. Salinity and suspended matter influenced the dominant zooplankton taxa at E2 and E1, respectively. Zooplankton assemblages exhibited a pattern of prominent diurnal-spatial variation in comparison to the tidal scale in the Mahanadi Estuary. Overall, this study documented a very high zooplankton diversity (92 taxa belonging to 13 groups) and significant variations in species abundance which highlighted the importance of carrying out sampling over the tidal cycle at contrasting salinity regimes.

In vivo and in vitro assessment of host-parasite interaction between the parasitic copepod Brachiella thynni (Copepoda; Lernaeopodidae) and the farmed Atlantic bluefin tuna (Thunnus thynnus).

The Atlantic bluefin tuna (ABFT; Thunnus thynnus) today represents one of the economically most important species for Croatian fisheries industry. Although the most diverse and abundant parasitofauna is usually found in the largest specimens of wild ABFT, the opposite was observed in captivity where parasite populations significantly decline by the end of the farming cycle. Copepod Brachiella thynni, is a skin parasite frequently parasitizing tuna, whose population also decreases in number throughout the rearing process. In order to better understand the immunity mechanisms underlying ABFT reaction to B. thynni infection, we studied expression profiles of immunity related genes; interleukin 1ß (il1ß), tumour necrosis factors (tnfα1, tnfα2), complement component 4 (c4) and caspase 3 (casp3), in peripheral blood leukocytes (PBLs) during in vitro stimulation by B. thynni protein extracts (i.e. antigens) and in infected tissues at B. thynni parasitation site. Finally, a histopathological analysis of semi-thin and ultra-thin sections of tissues surrounding B. thynii attachment site was performed to evaluate the severity of parasite-induced lesions and identify involved cell lineages. In vitro stimulation of ABFT PBLs with B. thynii antigens caused a dose-depended upregulation of selected genes, among which tnfα1 showed the highest induction by both concentrations of B. thynni protein extract. However, targeted genes were not significantly upregulated in the infected tissue. Also, no significant alterations in ultrastructure of epithelial layers surrounding B. thynii attachment site were noticed, except local tissue erosion, necrosis of squamous epithelium and proliferation of rodlet and goblet cells. Our results suggest that B. thynii has evolved strategies to successfully bypass both innate immune response and the connective-tissue proliferation processes. Therefore, the observed disappearance of this copepod by the end of the rearing process is more likely related to its limited lifespan on the host and its inability to complete the life cycle in the rearing cages, rather than host's reaction.

Pathology of multifocal purple spots, a nonspecific lesion morphology of Caribbean sea fans Gorgonia spp.

Disease is contributing to the decline of coral reefs globally, but the cause and pathogenesis of most coral diseases are poorly understood. Using Gorgonia ventalina and G. flabellum as a model for coral disease diagnosis, we histologically and microbiologically examined 45 biopsies of lesions resembling Gorgonia multifocal purple spots (MFPS) with the aim of forming a comprehensive case definition based on gross and microscopic morphologic descriptions and associated etiologies. Macroscopically, all lesions were small circular areas of purple pigmentation. Gross morphologies included pigmentation only (4/45, 9%), or pigmentation with branchlet expansion and fusion (19/45, 22%), sessile masses (17/45, 38%), or hard nodules (5/45, 9%). Histological morphologic diagnoses included amoebocyte encapsulation (9/45, 20%), coenenchymal amoebocytosis (6/45, 13%), melanin (17/45, 38%), and gorgonin deposition (13/45, 29%). Sixty-four percent of instances of fungi and 86% of labyrinthulomycetes were localized to grossly normal portions of the biopsy, whereas barnacles were only within lesions, and 87% of instances of algae and 82% of cyanobacteria were within lesioned area of the biopsy. Penicillium (n = 12) was the predominant genus of fungi isolated from biopsies. Barnacles were identified as Conopea sp. using molecular techniques. The pathology and etiology underlying MFPS lesions are diverse, consistent with a highly nonspecific lesion pattern rather than a specific disease. This study demonstrates the importance of microscopic examination of tissues for accurate classification of coral diseases and lesion patterns.

Morphology, molecular phylogeny, and taxonomy of trichodinids (Ciliophora, Mobilida) from Calanoid copepods.

Trichodinid ciliates were isolated from calanoid copepods and were characterized by morphological and molecular techniques. The trichodinids found in this study were morphologically similar to Trichodina acuta, but morphometric differences were observed between our specimens and T. acuta. The T. acuta specimens, isolated from fish in several geographical locations around the world, were slightly larger than specimens in our population. In fact, our morphological and morphometric data are more similar to Trichodina diaptomi populations, also found on copepods. Moreover, our phylogenetic analyses revealed that the 18S rRNA gene sequence from our samples emerged in a clade with two T. acuta sequences, infesting fish hosts in China. These three sequences showed 100% identity. Our data, along with a taxonomic review, suggest that the species T. diaptomi, described recently, is actually a synonym of T. acuta and that this species may be related to fish and copepod hosts. In conclusion, we highlight the urgent need to better investigate the polymorphism and host specificity within the Mobilida, as this data may represent important characters to better understand the evolution into this order.

The cleavage pattern of calanoid copepods-a case study.

Many crustacean groups show stereotyped cleavage patterns during early ontogeny. However, these patterns differ between the various major crustacean taxa, and a general mode is difficult to extract. Previous studies suggested that also copepods undergo an early cleavage with a more or less stereotyped pattern of blastomere divisions and fates. Yet, copepod embryology has been largely neglected. The last investigation of this kind dates back more than a century and the results are somewhat contradictory when compared with those of other researchers. To overcome these problems, we studied the early development of a so far undescribed calanoid copepod species, Skistodiaptomus sp., applying histochemical staining, confocal laser scanning microscopy, and bifocal 4D microscopy. The blastomere arrangement of the four-cell stage of this species varies to a large degree. It can either form a typical radial pattern with the four blastomeres lying in one plane or a tilted orientation of the axes connecting the sister cells of the previous division. In both cases, a stereotyped division pattern is maintained inside each quadrant during subsequent cleavages. In addition, we found two types of blastomere arrangements with a mirror symmetry. Most divisions within the quadrants follow the perpendicularity rule until the eighth cleavage. Deviations from this rule occur only in the narrow regions where the different quadrants touch and near the site of gastrulation. Gastrulation is initiated around the descendants of one individually identifiable blastomere of the 16-cell stage. This cell divides in a specific manner forming a characteristic cell arrangement, the gastrulation triangle. This gastrulation triangle initiates the internalization process of the gastrulation and it is encircled by another characteristic cell type, the crown cells. Our observations reveal several similarities to the early development of Calanus finmarchicus, another calanoid species. These relate to blastomere arrangements and divisions and the pattern of gastrulation. As Calanoida represent a basal or near basal branch of the copepod tree, this description will provide the ground for reconstruction of the cleavage pattern of the last common ancestor of Copepoda.

A New Species of Lagochondria (Crustacea: Copepoda) Parasitic on Callionymid Fish: First Record of Precopulatory Mate Guarding in Fish-Parasitic Cyclopoid Copepods, and of the Occurrence of the Genus in the North Pacific.

A new species of the genus Lagochondria of the parasitic cyclopoid family Chondracanthidae is described from the gill cavity of the callionymid Repomucenus virgis (Jordan and Fowler, 1903) collected from off Kii Peninsula, Japan. This is the first record of the occurrence of the genus from the North Pacific, and is only the second species in the genus. The female of the new species is easily distinguished from that of the Australian type species L. nana Ho and Dojiri, 1988 by having a squared trunk lacking paired posterior processes, and by the very short neck. In the male, the new species can be differentiated from the type species by having three inner setae on the caudal ramus, and by the distal segment of the antennule having a setal formula of 4, 3, 7+ ae. An adult female was accompanied by an attached adult male, whereas fourth and fifth copepodid females each carried a fifth and a fourth attached copepodid male, respectively. This is the first record of precopulatory mate guarding in a cyclopoid family parasitic on fish hosts, and of mate guarding between late copepodids of both sexes. The zoogeography of the genus and its relatives with an atrophied tip on the antenna is also discussed.

The Asian cyclopoid copepod Mesocyclops pehpeiensis reported from the western basin of Lake Erie.

The Asian cyclopoid copepod Mesocyclops pehpeiensis Hu, 1943 has been reported as an introduced species at several locations in the western hemisphere. In the United States, reports of this exotic species are restricted to localities in Louisiana, Mississippi, and Washington D.C. This report documents a new record of occurrence for M. pehpeiensis from the western basin of Lake Erie. The detection of M. pehpeiensis in Lake Erie constitutes the first record of this species from the Laurentian Great Lakes, and the northernmost record in the western hemisphere. The species was found in 2016, 2017 and 2018, including females with egg sacks, and can therefore be considered established in the area. The occurrence of M. pehpeiensis in Lake Erie suggests that this Asian copepod may be more widely distributed in North America than is currently understood.

Evolutionary History of Chemosensory-Related Gene Families across the Arthropoda.

Chemosensory-related gene (CRG) families have been studied extensively in insects, but their evolutionary history across the Arthropoda had remained relatively unexplored. Here, we address current hypotheses and prior conclusions on CRG family evolution using a more comprehensive data set. In particular, odorant receptors were hypothesized to have proliferated during terrestrial colonization by insects (hexapods), but their association with other pancrustacean clades and with independent terrestrial colonizations in other arthropod subphyla have been unclear. We also examine hypotheses on which arthropod CRG family is most ancient. Thus, we reconstructed phylogenies of CRGs, including those from new arthropod genomes and transcriptomes, and mapped CRG gains and losses across arthropod lineages. Our analysis was strengthened by including crustaceans, especially copepods, which reside outside the hexapod/branchiopod clade within the subphylum Pancrustacea. We generated the first high-resolution genome sequence of the copepod Eurytemora affinis and annotated its CRGs. We found odorant receptors and odorant binding proteins present only in hexapods (insects) and absent from all other arthropod lineages, indicating that they are not universal adaptations to land. Gustatory receptors likely represent the oldest chemosensory receptors among CRGs, dating back to the Placozoa. We also clarified and confirmed the evolutionary history of antennal ionotropic receptors across the Arthropoda. All antennal ionotropic receptors in E. affinis were expressed more highly in males than in females, suggestive of an association with male mate-recognition behavior. This study is the most comprehensive comparative analysis to date of CRG family evolution across the largest and most speciose metazoan phylum Arthropoda.

Lack of host specificity of copepod crustaceans associated with mushroom corals in the Red Sea.

The radiation of symbiotic copepods (Crustacea: Copepoda) living in association with stony corals (Cnidaria: Scleractinia) is considered host-specific and linked to the phylogenetic diversification of their hosts. However, symbiotic copepods are poorly investigated, occurrence records are mostly anecdotal, and no explicit analysis exists regarding their relationship with the hosts. Here, we analysed the occurrence of symbiotic copepods on different co-occurring and phylogenetically closely related scleractinian corals. We used an innovative approach of DNA extraction from single microscopic specimens that preserves the shape of the organisms for integrative morphological studies. The rationale of the study involved: (i) sampling of mushroom corals (Fungiidae) belonging to 13 species and eight genera on different reefs along the Saudi coastline in the Red Sea, (ii) extraction of all the associated copepods, (iii) morphological screening and identification of copepod species, (iv) use of DNA taxonomy on mitochondrial and nuclear markers to determine species boundaries for morphologically unknown copepod species, (v) reconstruction of phylogenies to understand their evolutionary relationships, and (vi) analysis of the ecological drivers of the occurrence, diversity and host specificity of the copepods. The seven species of coral-associated copepods, all new to science, did not show any statistically significant evidence of host-specificity or other pattern of ecological association. We thus suggest that, contrary to most assumptions and previous anecdotal evidence on this coral-copepod host-symbiont system, the association between copepods and their host corals is rather labile, not strict, and not phylogenetically constrained, changing our perception on evolutionary patterns and processes in symbiotic copepods.

Swim and fly: escape strategy in neustonic and planktonic copepods.

Copepods can respond to predators by powerful escape jumps that in some surface-dwelling forms may propel the copepod out of the water. We studied the kinematics and energetics of submerged and out-of-water jumps of two neustonic pontellid copepods, Anomalocera patersoni and Pontella mediterranea, and one pelagic calanoid copepod, Calanus helgolandicus (euxinus). We show that jumping out of the water does not happen just by inertia gained during the copepod's acceleration underwater, but also requires the force generated by the thoracic limbs when breaking through the water's surface to overcome surface tension, drag and gravity. The timing of this appears to be necessary for success. At the moment of breaking the water interface, the instantaneous velocity of the two pontellids reached 125 cm s-1, while their maximum underwater speed (115 cm s-1) was close to that of similarly sized C. helgolandicus (106 cm s-1). The average specific power produced by the two pontellids during out-of-water jumps (1700-3300 W kg-1 muscle mass) was close to that during submerged jumps (900-1600 W kg-1 muscle mass) and, in turn, similar to that produced during submerged jumps of C. helgolandicus (1300 W kg-1 muscle mass). The pontellids may shake off water adhering to their body by repeated strokes of the limbs during flight, which leads to a slight acceleration in the air. Our observations suggest that out-of-water jumps of pontellids are not dependent on any exceptional ability to perform this behavior but have the same energetic cost and are based on the same kinematic patterns and contractive capabilities of muscles as those of copepods swimming submerged.

Oomycete parasites in freshwater copepods of Patagonia: effects on survival and recruitment.

Copepods are hosts to various oomycete parasite species, but the effects of pathogens on copepod populations have rarely been studied. This study aimed to characterize oomycete infection in the freshwater copepod Parabroteas sarsi in a temporary pond in Patagonia (Argentina). A complete hydroperiod was monitored, evaluating environmental variables as influencing factors in the oomycete infections. Laboratory experiments were performed to evaluate the susceptibility of infected copepods to consumption by predators. Although 5 species of copepods were present in the pond, only ovigerous P. sarsi females were parasitized by oomycetes. Two species of oomycetes were always found together: Aphanomyces ovidestruens and Pythium flevoense. Infections were detected at water temperatures >20°C, with a positive relationship between temperature and parasite prevalence. Infection occurred after a decrease in large filter-feeder densities. The pathogens were not lethal to P. sarsi females in the short-term, but did produce mortality of entire egg sacs, thus negatively impacting subsequent recruitment. Mean prevalence of infection in females was 53%, reaching 83% in December. Females have the capacity to release an infected egg sac and generate a new one in a few days. This infection does not affect the susceptibility of P. sarsi to the predator Notonecta vereertbruggheni. The decrease in female abundance registered towards the end of the hydroperiod could be related to a combination of factors that may have a differential effect on female survival, such as increasing temperature, the energy cost of egg sac development and carrying and oomycete infection.

Impact of habitat heterogeneity on zooplankton assembly in a temperate river-floodplain system.

Dissimilar life features of Rotifera, Cladocera and Copepoda enable these organisms to respond differently to changes in the hydrological regime which influence alterations in environmental characteristics. We investigated the effect of habitat heterogeneity (e.g. eupotamal, parapotamal, palaeopotamal) on individual zooplankton group assemblages and biodiversity indices (α, ß and γ diversity) during hydro regime change in floodplain waterbodies. Dissolved oxygen and organic nitrogen concentrations changed significantly among hydrological states while water depth was affected by both site and hydro regime replacement. Each studied site supported different zooplankton assemblage that highly depended on species-specific responses to hydro regime change. Also, individual zooplankton groups exhibited different correlations with specific environmental parameters regarding site change. Throughout the study, rotifers' local (α) and among-community (ß) diversities were susceptible to the site and inundation change while the microcrustacean biodiversity pattern diverged. Copepods highly discriminated different habitat types and hydrological phases at the regional scale (γ diversity), while we found a complete lack of biodiversity dependence on both site and hydrology for Cladocera. Our results show that heterogeneous environments support the development of different zooplankton assemblages that express the within-group dissimilarities. They also point to the importance of identifying processes in hydrologically variable ecosystems that influence biodiversity patterns at an individual zooplankton group level. Our results suggest the use of appropriate zooplankton groups as biological markers in natural habitats and stress the importance of proper management in preserving biodiversity in floodplain areas.

Phylogenomic analysis of Copepoda (Arthropoda, Crustacea) reveals unexpected similarities with earlier proposed morphological phylogenies.

BACKGROUND: Copepods play a critical role in marine ecosystems but have been poorly investigated in phylogenetic studies. Morphological evidence supports the monophyly of copepods, whereas interordinal relationships continue to be debated. In particular, the phylogenetic position of the order Harpacticoida is still ambiguous and inconsistent among studies. Until now, a small number of molecular studies have been done using only a limited number or even partial genes and thus there is so far no consensus at the order-level. RESULTS: This study attempted to resolve phylogenetic relationships among and within four major copepod orders including Harpacticoida and the phylogenetic position of Copepoda among five other crustacean groups (Anostraca, Cladocera, Sessilia, Amphipoda, and Decapoda) using 24 nuclear protein-coding genes. Phylogenomics has confirmed the monophyly of Copepoda and Podoplea. However, this study reveals surprising differences with the majority of the copepod phylogenies and unexpected similarities with postembryonic characters and earlier proposed morphological phylogenies; More precisely, Cyclopoida is more closely related to Siphonostomatoida than to Harpacticoida which is likely the most basally-branching group of Podoplea. Divergence time estimation suggests that the origin of Harpacticoida can be traced back to the Devonian, corresponding well with recently discovered fossil evidence. Copepoda has a close affinity to the clade of Malacostraca and Thecostraca but not to Branchiopoda. This result supports the hypothesis of the newly proposed clades, Communostraca, Multicrustacea, and Allotriocarida but further challenges the validity of Hexanauplia and Vericrustacea. CONCLUSIONS: The first phylogenomic study of Copepoda provides new insights into taxonomic relationships and represents a valuable resource that improves our understanding of copepod evolution and their wide range of ecological adaptations.

Effects of the vertically transmitted microsporidian Facilispora margolisi and the parasiticide emamectin benzoate on salmon lice (Lepeophtheirus salmonis).

BACKGROUND: Microsporidia are highly specialized, parasitic fungi that infect a wide range of eukaryotic hosts from all major taxa. Infections cause a variety of damaging effects on host physiology from increased stress to death. The microsporidian Facilispora margolisi infects the Pacific salmon louse (Lepeophtheirus salmonis oncorhynchi), an economically and ecologically important ectoparasitic copepod that can impact wild and cultured salmonids. RESULTS: Vertical transmission of F. margolisi was demonstrated by using PCR and in situ hybridization to identify and localize microsporidia in female L. salmonis and their offspring. Spores and developmental structures of F. margolisi were identified in 77% of F1 generation copepods derived from infected females while offspring from uninfected females all tested negative for the microsporidia. The transcriptomic response of the salmon louse to F. margolisi was profiled at both the copepodid larval stage and the pre-adult stage using microarray technology. Infected copepodids differentially expressed 577 transcripts related to stress, ATP generation and structural components of muscle and cuticle. The infection also impacted the response of the copepodid to the parasiticide emamectin benzoate (EMB) at a low dose of 1.0 ppb for 24 h. A set of 48 transcripts putatively involved in feeding and host immunomodulation were up to 8-fold underexpressed in the F. margolisi infected copepodids treated with EMB compared with controls or either stressor alone. Additionally, these infected lice treated with EMB also overexpressed 101 transcripts involved in stress resistance and signalling compared to the other groups. In contrast, infected pre-adult lice did not display a stress response, suggesting a decrease in microsporidian virulence associated with lice maturity. Furthermore, copepodid infectivity and moulting was not affected by the microsporidian infection. CONCLUSIONS: This study demonstrated that F. margolisi is transmitted vertically between salmon louse generations and that biological impacts of infection differ depending on the stage of the copepod host. The infection caused significant perturbations of larval transcriptomes and therefore must be considered in future studies in which impacts to host development and environmental factors are assessed. Fitness impacts are probably minor, although the interaction between pesticide exposure and microsporidian infection merits further study.

Enhanced transcriptomic responses in the Pacific salmon louse Lepeophtheirus salmonis oncorhynchi to the non-native Atlantic Salmon Salmo salar suggests increased parasite fitness.

BACKGROUND: Outcomes of infections with the salmon louse Lepeophtheirus salmonis vary considerably among its natural hosts (Salmo, Oncorhynchus spp.). Host-parasite interactions range from weak to strong host responses accompanied by high to low parasite abundances, respectively. Parasite behavioral studies indicate that the louse prefers the host Atlantic Salmon (Salmo salar), which is characterized by a weak immune response, and that this results in enhanced parasite reproduction and growth rates. Furthermore, parasite-derived immunosuppressive molecules (e.g., proteases) have been detected at higher amounts in response to the mucus of Atlantic Salmon relative to Coho Salmon (Oncorhynchus kisutch). However, the host-specific responses of the salmon louse have not been well characterized in either of the genetically distinct sub-species that occur in the Atlantic and Pacific Oceans. RESULTS: We assessed and compared the transcriptomic feeding response of the Pacific salmon louse (L. salmonis oncorhynchi,) while parasitizing the highly susceptible Atlantic Salmon and Sockeye Salmon (Oncorhynchus nerka) or the more resistant Coho Salmon (Oncorhynchus kisutch) using a 38 K oligonucleotide microarray. The response of the louse was enhanced both in the number of overexpressed genes and in the magnitude of expression while feeding on the non-native Atlantic Salmon, compared to either Coho or Sockeye Salmon. For example, putative virulence factors (e.g., cathepsin L, trypsin, carboxypeptidase B), metabolic enzymes (e.g., cytochrome B, cytochrome C), protein synthesis enzymes (e.g., ribosomal protein P2, 60S ribosomal protein L7), and reproduction-related genes (e.g., estrogen sulfotransferase) were overexpressed in Atlantic-fed lice, indicating heightened parasite fitness with this host species. In contrast, responses in Coho- or Sockeye-fed lice were more similar to those of parasites deprived of a host. To test for host acclimation by the parasite, we performed a reciprocal host transfer experiment and determined that the exaggerated response to Atlantic Salmon was independent of the initial host species, confirming our conclusion that the Pacific salmon louse exhibits an enhanced response to Atlantic Salmon. CONCLUSIONS: This study characterized global transcriptomic responses of Pacific salmon lice during infection of susceptible and resistant hosts. Similar parasite responses during infection of Coho or Sockeye Salmon, despite differences in natural immunity to infection between these host species, indicate that host susceptibility status alone does not drive the parasite response. We identified an enhanced louse response after feeding on Atlantic Salmon, characterized by up-regulation of virulence factors, energy metabolism and reproductive-associated transcripts. In contrast, the responses of lice infecting Coho or Sockeye Salmon were weaker, with reduced expression of virulence factors. These observations indicate that the response of the louse is independent of host susceptibility and suggest that co-evolutionary host-parasite relationships may influence contemporary host-parasite interactions. This research improves our understanding of the susceptibility of Atlantic Salmon and may assist in the development of novel control measures against the salmon louse.