Special Issue "Emerging Viruses in Aquaculture".

According to the 2018 FAO report on aquaculture, there are 598 species of finfish, molluscs, crustaceans, and other organisms used in aquafarming around the world [...].

Mini Review: Virus Interference: History, Types and Occurrence in Crustaceans.

Virus interference is a phenomenon in which two viruses interact within a host, affecting the outcome of infection of at least one of such viruses. The effect of this event was first observed in the XVIII century and it was first recorded even before virology was recognized as a distinct science from microbiology. Studies on virus interference were mostly done in the decades between 1930 and 1960 in viruses infecting bacteria and different vertebrates. The systems included in vivo experiments and later, more refined assays were done using tissue and cell cultures. Many viruses involved in interference are pathogenic to humans or to economically important animals. Thus the phenomenon may be relevant to medicine and to animal production due to the possibility to use it as alternative to chemical therapies against virus infections to reduce the severity of disease/mortality caused by a superinfecting virus. Virus interference is defined as the host resistance to a superinfection caused by a pathogenic virus causing obvious signs of disease and/or mortality due to the action of an interfering virus abrogating the replication of the former virus. Different degrees of inhibition of the superinfecting virus can occur. Due to the emergence of novel pathogenic viruses in recent years, virus interference has recently been revisited using different pathogens and hosts, including commercially important farmed aquatic species. Here, some highly pathogenic viruses affecting farmed crustaceans can be affected by interference with other viruses. This review presents data on the history of virus interference in hosts including bacteria and animals, with emphasis on the known cases of virus interference in crustacean hosts. Life Science Identifiers (LSIDs) Escherichia coli [(Migula 1895) Castellani & Chalmers 1919] Aedes albopictus (Skuse 1894) Liocarcinus depurator (Linnaeus 1758): urn:lsid:marinespecies.org:taxname:107387 Penaeus duorarum (Burkenroad 1939): urn:lsid:marinespecies.org:taxname:158334 Carcinus maenas (Linnaeus 1758): urn:lsid:marinespecies.org:taxname:107381 Macrobrachium rosenbergii (De Man 1879): urn:lsid:marinespecies.org:taxname:220137 Penaeus vannamei (Boone 1931): urn:lsid:zoobank.org:pub:C30A0A50-E309-4E24-851D-01CF94D97F23 Penaeus monodon (Fabricius 1798): urn:lsid:zoobank.org:act:3DD50D8B-01C2-48A7-B80D-9D9DD2E6F7AD Penaeus stylirostris (Stimpson 1874): urn:lsid:marinespecies.org:taxname:584982.

Diversity and classification of reoviruses in crustaceans: A proposal.

A variety of reoviruses have been described in crustacean hosts, including shrimp, crayfish, prawn, and especially in crabs. However, only one genus of crustacean reovirus - Cardoreovirus - has been formally recognized by ICTV (International Committee on Taxonomy of Viruses) and most crustacean reoviruses remain unclassified. This arises in part from ambiguous or incomplete information on which to categorize them. In recent years, increased availability of crustacean reovirus genomic sequences is making the discovery and classification of crustacean reoviruses faster and more certain. This minireview describes the properties of the reoviruses infecting crustaceans and suggests an overall classification of brachyuran crustacean reoviruses based on a combination of morphology, host, genome organization pattern and phylogenetic sequence analysis.

Structure analysis of thymidylate synthase from white spot syndrome virus reveals WSSV-specific structural elements.

White spot syndrome virus (WSSV), the causative agent of white spot disease (WSD) severely affecting crustacean life forms, is highly contagious and forms the principal cause of massive economic losses in the shrimp aquaculture industry. Previous studies have demonstrated thymidylate synthase as a successful anti-cancer therapeutic drug target, leading to various anti-cancer drugs. The differential utilization of nucleotide precursors between white spot syndrome virus and shrimp encouraged us to analyze WSSV-thymidylate synthase (wTS). Here, we report the crystal structures of wTS in its apo-form and as a ternary complex with deoxyuridine monophosphate (dUMP) and methotrexate at a resolution of 2.35 Å and 2.6 Å, respectively. wTS possesses a fold characteristic to known thymidylate synthase (TS) structures. Like other TS structures, the apo-form of wTS displays an open conformation, whereas the wTS ternary complex attains a closed conformation. While the C-terminal loop maintains a typical distance from methotrexate, the Sγ atom of the catalytic Cys is positioned farther from the C6 atom of dUMP. Altogether, we report the first TS structure from a crustacean virus and highlight its distinction from shrimp and other TS structures.

Simultaneous detection of multiple bacterial and viral aquatic pathogens using a fluorogenic loop-mediated isothermal amplification-based dual-sample microfluidic chip.

Rapid and user-friendly diagnostic tests are necessary for early diagnosis and immediate detection of diseases, particularly for on-site screening of pathogenic microorganisms in aquaculture. In this study, we developed a dual-sample microfluidic chip integrated with a real-time fluorogenic loop-mediated isothermal amplification assay (dual-sample on-chip LAMP) to simultaneously detect 10 pathogenic microorganisms, that is Aeromonas hydrophila, Edwardsiella tarda, Vibrio harveyi, V. alginolyticus, V. anguillarum, V. parahaemolyticus, V. vulnificus, infectious hypodermal and haematopoietic necrosis virus, infectious spleen and kidney necrosis virus, and white spot syndrome virus. This on-chip LAMP provided a nearly automated protocol that can analyse two samples simultaneously, and the tests achieved limits of detection (LOD) ranging from 100 to 10-1  pg/µl for genomic DNA of tested bacteria and 10-4 to 10-5  pg/µl for recombinant plasmid DNA of tested viruses, with run times averaging less than 30 min. The coefficient of variation for the time-to-positive value was less than 10%, reflecting a robust reproducibility. The clinical sensitivity and specificity were 93.52% and 85.53%, respectively, compared to conventional microbiological or clinical methods. The on-chip LAMP assay provides an effective dual-sample and multiple pathogen analysis, and thus would be applicable to on-site detection and routine monitoring of multiple pathogens in aquaculture.

Research progress on viral accommodation 2009 to 2019.

The viral accommodation hypothesis for crustaceans and insects was first proposed in 1998/2001, stimulated by observations that shrimp and insects or insect cell lines can coexist with both DNA or RNA viruses without showing any signs of disease (i.e., they tolerate, single to multiple, persistent infections, sometimes for a lifetime). A review of tests of the hypothesis up to 2007 was previously published in DCI. This was followed by a major revision in 2009 when the elusive memory element required by the hypothesis was proposed to reside in non-retroviral fragments of extant viruses, now called endogenous viral elements (EVE) that are autonomously inserted into the host genome as cDNA copied from viral mRNA. Here, progress in research on viral accommodation in crustaceans and insects over the decade following 2009 is reviewed. It culminates with a discussion of exiting research results from insects in 2019 that prove the existence of specific, adaptive and heritable immunity, at least in mosquitoes. It remains to be determined whether the same mechanisms also govern EVE acquisition and its protective RNA production in shrimp. The wide-ranging consequences of the revealed mechanisms for viral disease control in economic crustaceans and insects is discussed.

The Anti-lipopolysaccharide Factors in Crustaceans.

Anti-lipopolysaccharide factors (ALFs) are a type of antimicrobial peptide (AMP) which show broad-spectrum antimicrobial activity against Gram-positive bacteria, Gram-negative bacteria, fungi and viruses. In this chapter, we review the discovery and classification of this kind of antimicrobial peptide in crustaceans. The structure and function, as well as the mechanism of antibacterial and antiviral activities of ALFs will be summarized and discussed. We will then describe the expression and regulation of various ALF genes in different crustacean species. Finally, the application prospects of ALFs in drug development and disease-resistant genetic breeding will be pointed out and discussed. The review will also discuss several key questions such as the systematic classification and expression regulation of the ALF genes, as well as the future application of ALFs and ALF-derived peptides.

A New Family of DNA Viruses Causing Disease in Crustaceans from Diverse Aquatic Biomes.

Panulirus argus virus 1 (PaV1) is the only known virus infecting the Caribbean spiny lobster (Panulirus argus) from the Caribbean Sea. Recently, related viruses, Dikerogammarus haemobaphes virus 1 (DhV1) and Carcinus maenas virus 1 (CmV1), have been detected in the demon shrimp (Dikerogammarus haemobaphes) and the European shore crab (Carcinus maenas), respectively, from sites in the United Kingdom. The virion morphology of these crustacean viruses is similar to that of iridoviruses. However, unlike iridoviruses and other nucleocytoplasmic large DNA viruses (NCLDVs), these viruses complete their morphogenesis in the host cell nucleus rather than in the cytoplasm. To date, these crustacean viruses have remained unclassified due to a lack of genomic data. Using an Illumina MiSeq sequencer, we sequenced the complete genomes of PaV1, CmV1, and DhV1. Comparative genome analysis shows that these crustacean virus genomes encode the 10 hallmark proteins previously described for the NCLDVs of eukaryotes, strongly suggesting that they are members of this group. With a size range of 70 to 74 kb, these are the smallest NCLDV genomes identified to date. Extensive gene loss, divergence of gene sequences, and the accumulation of low-complexity sequences reflect the extreme degradation of the genomes of these "minimal" NCLDVs rather than any direct relationship with the NCLDV ancestor. Phylogenomic analysis supports the classification of these crustacean viruses as a distinct family, "Mininucleoviridae," within the pitho-irido-Marseille branch of the NCLDVs.IMPORTANCE Recent genomic and metagenomic studies have led to a dramatic expansion of the known diversity of nucleocytoplasmic large DNA viruses (NCLDVs) of eukaryotes, which include giant viruses of protists and important pathogens of vertebrates, such as poxviruses. However, the characterization of viruses from nonmodel hosts still lags behind. We sequenced the complete genomes of three viruses infecting crustaceans, the Caribbean spiny lobster, demon shrimp, and European shore crab. These viruses have the smallest genomes among the known NCLDVs, with losses of many core genes, some of which are shared with iridoviruses. The deterioration of the transcription apparatus is compatible with microscopic and ultrastructural observations indicating that these viruses replicate in the nucleus of infected cells rather than in the cytoplasm. Phylogenomic analysis indicates that these viruses are sufficiently distinct from all other NCLDVs to justify the creation of a separate family, for which we propose the name "Mininucleoviridae" (i.e., small viruses reproducing in the cell nucleus).

ICTV Virus Taxonomy Profile: Nudiviridae.

Members of the family Nudiviridae are large dsDNA viruses with distinctive rod-shaped nucleocapsids and circular genomes of 96-232 kbp. Nudiviruses have been identified from a diverse range of insects and crustaceans and are closely related to baculoviruses. This is a summary of the International Committee on Taxonomy of Viruses Report on the taxonomy of the family Nudiviridae, which is available at ictv.global/report/nudiviridae.

Codon Usage in the Iflaviridae Family Is Not Diverse Though the Family Members Are Isolated from Diverse Host Taxa.

All iflavirus members belong to the unique genus, Iflavirus, of the family, Iflaviridae. The host taxa and sequence identities of these viruses are diverse. A codon usage bias, maintained by a balance between selection, mutation, and genetic drift, exists in a wide variety of organisms. We characterized the codon usage patterns of 44 iflavirus genomes that were isolated from the classes, Insecta, Arachnida, Mammalia, and Malacostraca. Iflaviruses lack a strong codon usage bias when they are evaluated using an effective number of codons. The odds ratios of the majority of dinucleotides are within the normal range. However, the dinucleotides at the 1st-2nd codon positions are more biased than those at the 2nd-3rd codon positions. Plots of effective numbers of codons, relative neutrality analysis, and PR2 bias analysis all indicate that selection pressure dominates mutations in shaping codon usage patterns in the family, Iflaviridae. When these viruses were grouped into their host taxa, we found that the indices, including the nucleotide composition, effective number of codons, relative synonymous codon usage, and the influencing factors behind the codon usage patterns, all show that there are non-significant differences between the six host-taxa-groups. Our results disagree with our assumption that diverse viruses should possess diverse codon usage patterns, suggesting that the nucleotide composition and codon usage in the family, Iflaviridae, are not host taxa-specific signatures.