Molecular detection of betanodaviruses from apparently healthy wild marine invertebrates

Gomez, Dennis K.; Baeck, Gun Wook; Kim, Ji Hyung; Choresca, Casiano H.; Park, Taesung

doi:10.1016/j.jip.2007.10.012

Cited by 45 publications

(41 citation statements)

References 22 publications

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“…However, TNCL virus was detected in AcMNPV-infected High Five cells (passage 98) used as a positive control. Persistent viral infections have been observed occasionally in insect cell lines [22,23] and nodaviruses are thought to occasionally infect wild marine invertebrates either latently or persistently [24]. A betanodavirus was also reported to be latently infecting a cell line derived from the brain tissue of the barramundi fish, Lates calcarifer [25].…”

Section: Discussionmentioning

confidence: 99%

Ao38, a new cell line from eggs of the black witch moth, Ascalapha odorata (Lepidoptera: Noctuidae), is permissive for AcMNPV infection and produces high levels of recombinant proteins

2010

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BackgroundThe insect cell line is a critical component in the production of recombinant proteins in the baculovirus expression system and new cell lines hold the promise of increasing both quantity and quality of protein production.ResultsSeventy cell lines were established by single-cell cloning from a primary culture of cells derived from eggs of the black witch moth (Ascalapha odorata; Lepidoptera, Noctuidae). Among 8 rapidly growing lines, cell line 38 (Ao38) was selected for further analysis, based on susceptibility to AcMNPV infection and production of secreted alkaline phosphatase (SEAP) from a baculovirus expression vector. In comparisons with low-passage High Five (BTI-Tn-5B1-4) cells, infected Ao38 cells produced β-galactosidase and SEAP at levels higher (153% and 150%, respectively) than those measured from High Five cells. Analysis of N-glycans of SEAP produced in Ao38 cells revealed two N-glycosylation sites and glycosylation patterns similar to those reported for High Five and Sf9 cells. Glycopeptide isoforms consisted of pauci- or oligomannose, with and without fucose on N-acetylglucosamine(s) linked to asparagine residues. Estimates of Ao38 cell volume suggest that Ao38 cells are approximately 2.5× larger than Sf9 cells but only approximately 74% of the size of High Five cells. Ao38 cells were highly susceptible to AcMNPV infection, similar to infectivity of Sf9 cells. Production of infectious AcMNPV budded virions from Ao38 cells peaked at approximately 4.5 × 107 IU/ml, exceeding that from High Five cells while lower than that from Sf9 cells. Ao38 cells grew rapidly in stationary culture with a population doubling time of 20.2 hr, and Ao38 cells were readily adapted to serum-free medium (Sf-900III) and to a suspension culture system. Analysis of Ao38 and a parental Ascalapha odorata cell line indicated that these lines were free of the alphanodavirus that was recently identified as an adventitious agent in High Five cell lines.ConclusionsAo38 cells represent a highly productive new insect cell line that will be useful for heterologous protein expression and other applications in biotechnology.

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Section: Discussionmentioning

confidence: 99%

Ao38, a new cell line from eggs of the black witch moth, Ascalapha odorata (Lepidoptera: Noctuidae), is permissive for AcMNPV infection and produces high levels of recombinant proteins

2010

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“…With the exception of a few metazoan parasites which also somewhat affect the Mytilus species [8]–[9], mussels seem refractory to diseases and could instead influence the prevalence of pathogens such as Perkinsus spp. and Betanodavirus (Nodaviridae) in other bivalves and fishes, respectively [10]. Like other invertebrates, bivalve molluscs rely on ancient and rapid defenses to fight potential pathogens, and gene-encoded antimicrobial peptides (AMPs) are major humoral components of their immune system.…”

Section: Introductionmentioning

confidence: 99%

Massively Parallel Amplicon Sequencing Reveals Isotype-Specific Variability of Antimicrobial Peptide Transcripts in Mytilus galloprovincialis

et al. 2011

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BackgroundEffective innate responses against potential pathogens are essential in the living world and possibly contributed to the evolutionary success of invertebrates. Taken together, antimicrobial peptide (AMP) precursors of defensin, mytilin, myticin and mytimycin can represent about 40% of the hemocyte transcriptome in mussels injected with viral-like and bacterial preparations, and unique profiles of myticin C variants are expressed in single mussels. Based on amplicon pyrosequencing, we have ascertained and compared the natural and Vibrio-induced diversity of AMP transcripts in mussel hemocytes from three European regions.Methodology/Principal FindingsHemolymph was collected from mussels farmed in the coastal regions of Palavas (France), Vigo (Spain) and Venice (Italy). To represent the AMP families known in M. galloprovincialis, nine transcript sequences have been selected, amplified from hemocyte RNA and subjected to pyrosequencing. Hemolymph from farmed (offshore) and wild (lagoon) Venice mussels, both injected with 107 Vibrio cells, were similarly processed. Amplicon pyrosequencing emphasized the AMP transcript diversity, with Single Nucleotide Changes (SNC) minimal for mytilin B/C and maximal for arthropod-like defensin and myticin C. Ratio of non-synonymous vs. synonymous changes also greatly differed between AMP isotypes. Overall, each amplicon revealed similar levels of nucleotidic variation across geographical regions, with two main sequence patterns confirmed for mytimycin and no substantial changes after immunostimulation.Conclusions/SignificanceBarcoding and bidirectional pyrosequencing allowed us to map and compare the transcript diversity of known mussel AMPs. Though most of the genuine cds variation was common to the analyzed samples we could estimate from 9 to 106 peptide variants in hemolymph pools representing 100 mussels, depending on the AMP isoform and sampling site. In this study, no prevailing SNC patterns related to geographical origin or Vibrio injection emerged. Whether or not the contact with potential pathogens can increase the amount of AMP transcript variants in mussels requires additional study.

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“…Based on phylogenetic analysis of the T4 variable region within the RNA2 segment, beta nodaviruses have been historically divided into 4 genotypes: striped jack nervous necrosis virus (SJNNV), tiger puffer nervous necrosis virus (TPNNV), barfin flounder nervous necrosis virus (BFNNV) and red-spotted grouper nervous necrosis virus (RGNNV) (Panzarin et al 2012). The phylo geny-based analyses of betanodaviruses previously reported in molluscs evidenced high homology with RGNNV (Gomez et al 2008a(Gomez et al , 2010 or RGNNV/ SJNNV reassortant strains (Panzarin et al 2012) circulating in fish in the areas of the studies. In our study, betanodaviruses were detected in skin, eyes and branchial-heart of both reared and wild common octopuses.…”

Section: Discussionmentioning

confidence: 68%

“…In fish, betanodavirus infection is known as viral nervous necrosis, fish encephalitis, viral encephalopathy and retinopathy (Shetty et al 2012). Betanodavirus infections have also been reported in marine invertebrates (Gomez et al 2008a,b, 2010, Panzarin et al 2012, including molluscs, such as the Mediterranean mussel Mytilus galloprovincialis Lamarck, 1819, the Japanese carpet shell Ruditapes philippinarum (Adams & Reeve, 1850) and the Japa nese common squid Todarodes pacificus (Steenstrup, 1880). Mediterranean mussels were found to be sub-clinically infected with betanodavirus, lo cated in the hepatopancreas (Gomez et al 2008a), while in Japanese common squid, betanodaviruses were detected in the brain and in eyes (Gomez et al 2008a(Gomez et al , 2010.…”

Section: Discussionmentioning

confidence: 99%

“…Betanodavirus infections have also been reported in marine invertebrates (Gomez et al 2008a,b, 2010, Panzarin et al 2012, including molluscs, such as the Mediterranean mussel Mytilus galloprovincialis Lamarck, 1819, the Japanese carpet shell Ruditapes philippinarum (Adams & Reeve, 1850) and the Japa nese common squid Todarodes pacificus (Steenstrup, 1880). Mediterranean mussels were found to be sub-clinically infected with betanodavirus, lo cated in the hepatopancreas (Gomez et al 2008a), while in Japanese common squid, betanodaviruses were detected in the brain and in eyes (Gomez et al 2008a(Gomez et al , 2010. Based on phylogenetic analysis of the T4 variable region within the RNA2 segment, beta nodaviruses have been historically divided into 4 genotypes: striped jack nervous necrosis virus (SJNNV), tiger puffer nervous necrosis virus (TPNNV), barfin flounder nervous necrosis virus (BFNNV) and red-spotted grouper nervous necrosis virus (RGNNV) (Panzarin et al 2012).…”

Section: Discussionmentioning

confidence: 99%

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Skin lesion-associated pathogens from Octopus vulgaris: first detection of Photobacterium swingsii, Lactococcus garvieae and betanodavirus

Fichi¹,

Cardeti²,

Perrucci³

et al. 2015

Dis. Aquat. Org.

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The common octopus Octopus vulgaris Cuvier, 1798 is extremely important in fisheries and is a useful protein source in most Mediterranean countries. Here we investigated pathogens associated with skin lesions in 9 naturally deceased specimens that included both cultured and wild common octopus. Within 30 min after death, each octopus was stored at 4°C and microbiologically examined within 24 h. Bacterial colonies, cultured from swabs taken from the lesions, were examined using taxonomical and biochemical analyses. Vibrio alginolyticus and V. parahaemolyticus were only isolated from cultured animals. A conventional PCR targeting the 16S ribosomal RNA (rRNA) gene and sequencing were performed on 2 bacterial isolates that remained unidentified after taxonomical and biochemical analysis. The sequence results indicated that the bacteria had a 99% identity with Lactococcus garvieae and Photobacterium swingsii. L. garvieae was confirmed using a specific PCR based on the 16S-23S rRNA internal transcribed spacer region, while P. swingsii was confirmed by phylogenetic analyses. Although all animals examined were found to be infected by the protozoan species Aggregata octopiana localised in the intestines, it was also present in skin lesions of 2 of the animals. Betanodavirus was detected in both cultured and wild individuals by cell culture, PCR and electron microscopy. These findings are the first report of L. garvieae and betanodavirus from skin lesions of common octopus and the first identification of P. swingsii both in octopus skin lesions and in marine invertebrates in Italy. Farto et al. (2003) isolated V. len tus from gills, branchial-heart and skin lesions of wild common octopus in Spain, confirming its pathogenic potential.Aggregata spp. (Protozoa: Apicomplexa) coccidian parasites are considered the dominant epizootiological agents in wild and cultured octopus stocks in Europe (Gestal et al. 2002a(Gestal et al. , 2007. In severe forms, Aggregata spp. infection spreads from the digestive tract to the mantle and the gills, affecting the connective tissue and epithelium of these organs and causing the appearance of white cysts associated with host cell hypertrophy with nuclear displacement, inflammation, phagocytosis, ulceration, and destruction of the organ architecture (Gestal et al. 2002b, CastellanosMartínez & Gestal 2013. A. octopiana is the only species of this genus reported in common octopus (Estévez et al. 1996, Castellanos-Martínez et al. 2013.Viruses infect most marine organisms, from bacteria to invertebrates and vertebrates (Pierce et al. 2010). Cephalopods, including common octopus, have been found to contain virus-like particles, probably of the genus Iridovirus, associated with tumours in the muscle tissue of the arms and, in advanced stages, on the funnel and the ventral surface of the mantle (Farley 1978, Hanlon & Forsythe 1990.The aim of the present study was to investigate the pathogens associated with skin lesions in cultured and wild common octopus. MATERIALS AND METHODS AnimalsBetw...

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Molecular detection of betanodaviruses from apparently healthy wild marine invertebrates

Cited by 45 publications

References 22 publications

Ao38, a new cell line from eggs of the black witch moth, Ascalapha odorata (Lepidoptera: Noctuidae), is permissive for AcMNPV infection and produces high levels of recombinant proteins

Ao38, a new cell line from eggs of the black witch moth, Ascalapha odorata (Lepidoptera: Noctuidae), is permissive for AcMNPV infection and produces high levels of recombinant proteins

Massively Parallel Amplicon Sequencing Reveals Isotype-Specific Variability of Antimicrobial Peptide Transcripts in Mytilus galloprovincialis

Skin lesion-associated pathogens from Octopus vulgaris: first detection of Photobacterium swingsii, Lactococcus garvieae and betanodavirus

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