High prevalence of Paramarteilia canceri infecting velvet swimming crabs Necora puber in Ireland.

The velvet swimming crab Necora puber has been fished in Ireland since the early 1980s and contributes significant income to smaller fishing vessels. From 2016 onwards, reduced landings have been reported. We undertook a full pathological investigation of crabs from fishing grounds at 3 sites on the west (Galway), southwest (Castletownbere) and east (Howth) coasts of Ireland. Histopathology, transmission electron microscopy and molecular taxonomic and phylogenetic analyses showed high prevalence and infection level of Paramarteilia canceri, previously only reported from the edible crab Cancer pagurus. This study provides the first molecular data for P. canceri, and shows its phylogenetic position in the order Paramyxida (Rhizaria). Other parasites and symbionts detected in the crabs were also noted, including widespread but low co-infection with Hematodinium sp. and a microsporidian consistent with the Ameson and Nadelspora genera. This is the first histological record of Hematodinium sp. in velvet crabs from Ireland. Four N. puber individuals across 2 sites were co-infected by P. canceri and Hematodinium sp. At one site, 3 velvet crabs infected with P. canceri were co-infected with the first microsporidian recorded from this host; the microsporidian 18S sequence was almost identical to Ameson pulvis, known to infect European shore crabs Carcinus maenas. The study provides a comprehensive phylogenetic analysis of this and all other available Ameson and Nadelspora 18S sequences. Together, these findings provide a baseline for further investigations of N. puber populations along the coast of Ireland.

Global mRNA and miRNA Analysis Reveal Key Processes in the Initial Response to Infection with WSSV in the Pacific Whiteleg Shrimp.

White Spot Disease (WSD) presents a major barrier to penaeid shrimp production. Mechanisms underlying White Spot Syndrome Virus (WSSV) susceptibility in penaeids are poorly understood due to limited information related to early infection. We investigated mRNA and miRNA transcription in Penaeus vannamei over 36 h following infection. Over this time course, 6192 transcripts and 27 miRNAs were differentially expressed-with limited differential expression from 3-12 h post injection (hpi) and a more significant transcriptional response associated with the onset of disease symptoms (24 hpi). During early infection, regulated processes included cytoskeletal remodelling and alterations in phagocytic activity that may assist WSSV entry and translocation, novel miRNA-induced metabolic shifts, and the downregulation of ATP-dependent proton transporter subunits that may impair cellular recycling. During later infection, uncoupling of the electron transport chain may drive cellular dysfunction and lead to high mortalities in infected penaeids. We propose that post-transcriptional silencing of the immune priming gene Dscam (downregulated following infections) by a novel shrimp miRNA (Pva-pmiR-78; upregulated) as a potential mechanism preventing future recognition of WSSV that may be suppressed in surviving shrimp. Our findings improve our understanding of WSD pathogenesis in P. vannamei and provide potential avenues for future development of prophylactics and treatments.

The first clawed lobster virus Homarus gammarus nudivirus (HgNV n. sp.) expands the diversity of the Nudiviridae.

Viral diseases of crustaceans are increasingly recognised as challenges to shellfish farms and fisheries. Here we describe the first naturally-occurring virus reported in any clawed lobster species. Hypertrophied nuclei with emarginated chromatin, characteristic histopathological lesions of DNA virus infection, were observed within the hepatopancreatic epithelial cells of juvenile European lobsters (Homarus gammarus). Transmission electron microscopy revealed infection with a bacilliform virus containing a rod shaped nucleocapsid enveloped in an elliptical membrane. Assembly of PCR-free shotgun metagenomic sequencing produced a circular genome of 107,063 bp containing 97 open reading frames, the majority of which share sequence similarity with a virus infecting the black tiger shrimp: Penaeus monodon nudivirus (PmNV). Multiple phylogenetic analyses confirm the new virus to be a novel member of the Nudiviridae: Homarus gammarus nudivirus (HgNV). Evidence of occlusion body formation, characteristic of PmNV and its closest relatives, was not observed, questioning the horizontal transmission strategy of HgNV outside of the host. We discuss the potential impacts of HgNV on juvenile lobster growth and mortality and present HgNV-specific primers to serve as a diagnostic tool for monitoring the virus in wild and farmed lobster stocks.

Shedding and survival of an intracellular pathogenic Endozoicomonas-like organism infecting king scallop Pecten maximus.

The Lyme Bay marine protected area (MPA) hosts a valuable population of king scallop Pecten maximus L. Recently, an Endozoicomonas-like organism (ELO), infecting host gill epithelial tissue, was associated with king scallop mass mortality events within the Lyme Bay MPA. Currently, very little is known about its transmission and survival outside the host. In this investigation, animals collected outside of reported mortality events showed high levels of ELO infection. Gill tissue disruption and the release of bacteria into the interlamellar space was seen histologically, suggesting shedding of ELO from host animals. To investigate pathogen survival outside the host, infected scallops were maintained in static water for a 24 h period, and then removed. Over the subsequent 8 d, water samples were collected and the quantity of ELO 16S rRNA transcript was measured by TaqManTM quantitative PCR (qPCR). The 16S rRNA transcript quantity was stable outside the host for 6 d before bacteria survival declined 2 logs (7.9 × 108 16S rRNA to 2.3 × 106 transcripts), suggesting that ELO can survive independently outside the host organism. The ELO-specific qPCR probe can therefore be used in future field studies of ELO prevalence within the environment and fauna of the Lyme Bay MPA.

Cucumispora ornata n. sp. (Fungi: Microsporidia) infecting invasive 'demon shrimp' (Dikerogammarus haemobaphes) in the United Kingdom.

Dikerogammarus haemobaphes, the 'demon shrimp', is an amphipod native to the Ponto-Caspian region. This species invaded the UK in 2012 and has become widely established. Dikerogammarus haemobaphes has the potential to introduce non-native pathogens into the UK, creating a potential threat to native fauna. This study describes a novel species of microsporidian parasite infecting 72.8% of invasive D. haemobaphes located in the River Trent, UK. The microsporidium infection was systemic throughout the host; mainly targeting the sarcolemma of muscle tissues. Electron microscopy revealed this parasite to be diplokaryotic and have 7-9 turns of the polar filament. The microsporidium is placed into the 'Cucumispora' genus based on host histopathology, fine detail parasite ultrastructure, a highly similar life-cycle and SSU rDNA sequence phylogeny. Using this data this novel microsporidian species is named Cucumispora ornata, where 'ornata' refers to the external beading present on the mature spore stage of this organism. Alongside a taxonomic discussion, the presence of a novel Cucumispora sp. in the United Kingdom is discussed and related to the potential control of invasive Dikerogammarus spp. in the UK and the health of native species which may come into contact with this parasite.