Transcriptomic evidence of a fourth mininucleovirus (Mininucleoviridae): A rapidly growing family among the Nucleo-Cytoplasmic Large DNA viruses (NCLDVs).

The Mininucleoviridae are crustacean-infecting viruses thought to drive mortality across aquatic biomes. Three have been characterised from Carcinus maenas, Panulirus argus, and Dikerogammarus haemobaphes. We screened 202 SRA datasets (NCBI) for novel mininucleoviruses from 44 amphipod species. Three metatranscriptome datasets from Gammarus lacustris contained sequences with similarity to Dikerogammarus haemobaphes mininucleovirus. Assembly resulted in 19 transcripts, 16 were putatively polycistronic. The putative Gammarus lacustris mininucleovirus shares 46 homologues with other mininucleoviruses (similarity range: 24.07 - 78.2 %). The transcripts from this putative virus highlight its likely association with the Mininucleoviridae.

A novel nudivirus infecting the invasive demon shrimp Dikerogammarus haemobaphes (Amphipoda).

The Nudiviridae are a family of large double-stranded DNA viruses that infects the cells of the gut in invertebrates, including insects and crustaceans. The phylogenetic range of the family has recently been enhanced via the description of viruses infecting penaeid shrimp, crangonid shrimp, homarid lobsters and portunid crabs. Here we extend this by presenting the genome of another nudivirus infecting the amphipod Dikerogammarus haemobaphes. The virus, which infects cells of the host hepatopancreas, has a circular genome of 119,754 bp in length, and encodes a predicted 106 open reading frames. This novel virus encodes all the conserved nudiviral genes (sharing 57 gene homologues with other crustacean-infecting nudiviruses) but appears to lack the p6.9 gene. Phylogenetic analysis revealed that this virus branches before the other crustacean-infecting nudiviruses and shares low levels of gene/protein similarity to the Gammanudivirus genus. Comparison of gene synteny from known crustacean-infecting nudiviruses reveals conservation between Homarus gammarus nudivirus and Penaeus monodon nudivirus; however, three genomic rearrangements in this novel amphipod virus appear to break the gene synteny between this and the ones infecting lobsters and penaeid shrimp. We explore the evolutionary history and systematics of this novel virus, suggesting that it be included in the novel Epsilonnudivirus genus (Nudiviridae).

Pathogens and other symbionts of the Amphipoda: taxonomic diversity and pathological significance.

With over 10000 species of Amphipoda currently described, this order is one of the most diverse groups of freshwater and marine Crustacea. Members of this group are globally distributed, and many are keystone species and ecosystem engineers within their respective ecologies. As with most organisms, disease is a key factor that can alter population size, behaviour, survival, invasion potential and physiology of amphipod hosts. This review explores symbiont diversity and pathology in amphipods by coalescing a range of current and historical literature to provide the first full review of our understanding of amphipod disease. The review is broken into 2 parts. The first half explores amphipod microparasites, which include data pertaining to viruses, bacteria, fungi, oomycetes, microsporidians, dinoflagellates, myxozoans, ascetosporeans, mesomycetozoeans, apicomplexans and ciliophorans. The second half reports the metazoan macroparasites of Amphipoda, including rotifers, trematodes, acanthocephalans, nematodes, cestodes and parasitic Crustacea. In all cases we have endeavoured to provide a complete list of known species that cause disease in amphipods, while also exploring the effects of parasitism. Although our understanding of disease in amphipods requires greater research efforts to better define taxonomic diversity and host effects of amphipod symbionts, research to date has made huge progress in cataloguing and experimentally determining the effects of disease upon amphipods. For the future, we suggest a greater focus on developing model systems that use readily available amphipods and diseases, which can be comparable to the diseases in other Crustacea that are endangered, economically important or difficult to house.

Pathogens of Dikerogammarus haemobaphes regulate host activity and survival, but also threaten native amphipod populations in the UK.

Dikerogammarus haemobaphes is a non-native amphipod in UK freshwaters. Studies have identified this species as a low-impact invader in the UK, relative to its cousin Dikerogammarus villosus. It has been suggested that regulation by symbionts (such as Microsporidia) could explain this difference in impact. The effect of parasitism on D. haemobaphes is largely unknown. This was explored herein using 2 behavioural assays measuring activity and aggregation. First, D. haemobaphes were screened histologically post-assay, identifying 2 novel viruses (D. haemobaphes bi-facies-like virus [DhbflV], D. haemobaphes bacilliform virus [DhBV]), Cucumispora ornata (Microsporidia), Apicomplexa, and Digenea, which could alter host behaviour. DhBV infection burden increased host activity, and C. ornata infection reduced host activity. Second, native invertebrates were collected from the invasion site at Carlton Brook, UK, and tested for the presence of C. ornata. PCR screening identified that Gammarus pulex and other native invertebrates were positive for C. ornata. The host range of this parasite, and its impact on host survival, was additionally explored using D. haemobaphes, D. villosus, and G. pulex in a laboratory trial. D. haemobaphes and G. pulex became infected by C. ornata, which also lowered survival rate. D. villosus did not become infected. A PCR protocol for DhbflV was also applied to D. haemobaphes after the survival trial, associating this virus with decreased host survival. In conclusion, D. haemobaphes has a complex relationship with parasites in the UK environment. C. ornata likely regulates populations by decreasing host survival and activity, but despite this benefit, the parasite threatens susceptible native wildlife.

Detection of viral hemorrhagic septicemia virus (VHSV) from Diporeia spp. (Pontoporeiidae, Amphipoda) in the Laurentian Great Lakes, USA.

The mode of viral hemorrhagic septicemia virus (VHSV) transmission in the Great Lakes basin is largely unknown. In order to assess the potential role of macroinvertebrates in VHSV transmission, Diporeia spp., a group of amphipods that are preyed upon by a number of susceptible Great Lakes fishes, were collected from seven locations in four of the Great Lakes and analyzed for the presence of VHSV. It was demonstrated that VHSV is present in some Diporeia spp. samples collected from lakes Ontario, Huron, and Michigan, but not from Lake Superior. Phylogenetic comparison of partial nucleoprotein (N) gene sequences (737 base pairs) of the five isolates to sequences of 13 other VHSV strains showed the clustering of Diporeia spp. isolates with the VHSV genotype IVb. This study reports the first incidence of a fish-pathogenic rhabdovirus being isolated from Diporeia, or any other crustacean and underscores the role macroinvertebrates may play in VHSV ecology.