scholarly journals Betanodavirus infection in Kuhlia rupestris and Ambassis marianus and isolation of betanodavirus from infected pond water

2020 ◽  
Vol 142 ◽  
pp. 1-11
Author(s):  
K Agnihotri ◽  
R Chong ◽  
D Underwood ◽  
C Kistler ◽  
M Hutchison
Keyword(s):  
Phylogenetic Analysis ◽  
Nucleotide Sequence ◽  
Cell Line ◽  
Environmental Samples ◽  
Sanger Sequencing ◽  
High Throughput Sequencing ◽  
Pond Water ◽  
Fish Cell ◽  
Nervous Necrosis Virus ◽  
Necrosis Virus

This is the first report of betanodavirus infection in 2 species of finfish, Kuhlia rupestris (jungle perch) and Ambassis marianus (estuary perchlet). This report also describes isolation of betanodavirus from infected pond water using the SSN-1 cell line. Histopathology of K. rupestris larvae revealed vacuolation in the eye and brain, which was confirmed using betanodavirus-specific immunohistochemistry. The eye and brain from A. marianus and betanodavirus isolated from pond water were confirmed using real-time PCR and Sanger sequencing. High throughput sequencing was used to obtain betanodavirus sequences from paraffin blocks containing infected K. rupestris. The phylogenetic analysis of betanodavirus RNA1 and RNA2 sequences from all 3 sources were associated with the red-spotted grouper nervous necrosis virus (RGNNV) genotype. The RNA1 nucleotide sequence from jungle perch showed 100% identity with the betanodavirus water isolate and 99.37% identity with A. marianus. Furthermore, we have demonstrated the usefulness of combining recovery of viable virus from environmental samples through fish cell line culture with PCR testing as a means of validating the efficacy of chlorination to eradicate betanodavirus from the pond environment.

scholarly journals Interference of the life cycle of fish nodavirus with fish retrovirus

2002 ◽  
Vol 83 (10) ◽  
pp. 2469-2474 ◽  
Author(s):  
K. W. Lee ◽  
S. C. Chi ◽  
T. M. Cheng
Keyword(s):  
Life Cycle ◽  
Cell Line ◽  
Culture Supernatant ◽  
Fish Cell ◽  
Nervous Necrosis Virus ◽  
Productivity Analysis ◽  
Necrosis Virus ◽  
Culture Supernatants ◽  
Titration System

Interference of the life cycle of grouper nervous necrosis virus (GNNV), a member of the Nodaviridae, genus Betanodavirus, by snakehead retrovirus (SnRV) has been studied in vitro. SGF-1, a new fish cell line that is persistently infected with SnRV, was induced by inoculating SnRV into the grouper fin cell line GF-1. Culture supernatants and cell pellets from both GNNV-infected SGF-1 and GF-1 cells were collected and employed for virus productivity analysis. The yields of GNNV RNA and capsid protein in GNNV-infected SGF-1 cells were similar to those in GNNV-infected GF-1 cells. However, when GF-1 cells were used for titration, the titre of the culture supernatant from GNNV-infected SGF-1 cells was much higher than that from GNNV-infected GF-1 cells. The titration result suggested that SnRV enhanced the infection or cytopathic effect (CPE) of GNNV during GNNV and SnRV coinfection of the GF-1 cell titration system, although SnRV cannot induce any CPE in GF-1 cells alone, nor can it increase the yield of GNNV after GNNV superinfection of SGF-1 cells. Moreover, GNNV cDNA was detected in both the pellet and the supernatant from GNNV-infected SGF-1 cells. This result indicated that SnRV reverse-transcribed the GNNV single-stranded genomic RNA into cDNA during GNNV superinfection of SGF-1 cells and created a new cDNA stage in the life cycle of the fish nodavirus.

scholarly journals Differential Nervous Necrosis Virus (NNV) Replication in Five Putative Susceptible Cell Lines

Pathogens ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 1565
Author(s):  
Yulema Valero ◽  
Carmen López-Vázquez ◽  
Sandra Souto ◽  
José G. Olveira ◽  
Alberto Cuesta ◽  
...  
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Control Measures ◽  
Effective Control ◽  
Fish Cell ◽  
Nervous Necrosis Virus ◽  
Necrosis Virus ◽  
Susceptible Cell ◽  
Viral Encephalopathy And Retinopathy

Viral encephalopathy and retinopathy caused by nervous necrosis virus (NNV), is one of the most threatening viral diseases affecting marine fish worldwide. In vitro propagation of NNV strains is essential for the design of effective control measures. In the present study we analysed both the susceptibility and the permissiveness of five fish cell lines (E-11, GF-1, SAF-1, DLB-1, and SaB-1) to three NNV strains (one RGNNV, one SJNNV, and one reassortant RGNNV/SJNNV). E-11 and DLB-1 were demonstrated to be highly susceptible to NNV strains, with average adsorption efficiency (AE) values higher than 90%. SAF-1 also showed high susceptibility (AE 88%), whereas GF-1 can be regarded as moderately susceptible (AE around 50%). On the contrary, SaB-1 can be considered a poorly susceptible cell line (AE values below 20%). E-11 and GF-1 cell lines provided the highest production rates for RGNNV and RG/SJ (around 103) and both cell lines can be regarded as fully permissive for these viral types. However, the SJNNV production rate in GF-1 was only 17.8 and therefore this cell line should be considered semi-permissive for this genotype. In SAF-1 cells, moderate viral replication was recorded but differences in intracellular and extracellular production suggest that viral progeny was not efficiently released. In DLB-1 and SaB-1 the final viral titres obtained in E-11 were lower than those of the inoculum. However, RNA1 synthesis values seem to indicate that RGNNV replication in DLB-1 and SAF-1 could have been underestimated, probably due to a poor adaptation of the virus grown in these cell lines to E-11. Based on all these results, E-11 seems to be the most appropriate cell for in vitro culture of RGNNV, SJNNV, and reassortant strains.

scholarly journals Identification and Sequence Analysis of a Novel Ilarvirus Infecting Sweet Cherry

Plants ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 514
Author(s):  
Chrysoula G. Orfanidou ◽  
Fei Xing ◽  
Jun Zhou ◽  
Shifang Li ◽  
Nikolaos I. Katis ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Nucleotide Sequence ◽  
Sanger Sequencing ◽  
Sweet Cherry ◽  
Novel Species ◽  
Peach Tree ◽  
Sweet Cherries ◽  
Blastn Analysis ◽  
Fruit Orchards ◽  
Prunus Spp

In the present study, we utilized high throughput and Sanger sequencing to determine the complete nucleotide sequence of a putative new ilarvirus species infecting sweet cherry, tentatively named prunus virus I (PrVI). The genome of PrVI is comprised of three RNA segments of 3474 nt (RNA1), 2911 nt (RNA2), and 2231 nt (RNA3) and features conserved motifs representative of the genus Ilarvirus. BlastN analysis revealed 68.1–71.9% nt identity of PrVI with strawberry necrotic shock virus (SNSV). In subsequent phylogenetic analysis, PrVI was grouped together with SNSV and blackberry chlorotic ringspot virus (BCRV), both members of subgroup 1 of ilarviruses. In addition, mini-scale surveys in stone fruit orchards revealed the presence of PrVI in a limited number of sweet cherries and in one peach tree. Overall, our data suggest that PrVI is a novel species of the genus Ilarvirus and it consists the fifth member of the genus that is currently known to infect Prunus spp.

scholarly journals Expression profile of selected genes of the E-11 cell line in response to red-spotted grouper nervous necrosis virus infection

2020 ◽  
Vol 18 ◽  
pp. 100468
Author(s):  
Kitipong Angsujinda ◽  
Timothy J. Mahony ◽  
Duncan R. Smith ◽  
Jes Kettratad ◽  
Wanchai Assavalapsakul
Keyword(s):  
Virus Infection ◽  
Cell Line ◽  
Expression Profile ◽  
Nervous Necrosis Virus ◽  
Necrosis Virus

scholarly journals Establishment of an infectious RNA transcription system for Striped jack nervous necrosis virus, the type species of the betanodaviruses

2001 ◽  
Vol 82 (11) ◽  
pp. 2653-2662 ◽  
Author(s):  
Tokinori Iwamoto ◽  
Kazuyuki Mise ◽  
Koh-ichiro Mori ◽  
Misao Arimoto ◽  
Toshihiro Nakai ◽  
...  
Keyword(s):  
Type Species ◽  
Fish Cell ◽  
Cdna Clones ◽  
Nervous Necrosis Virus ◽  
Necrosis Virus ◽  
T7 Promoter ◽  
Genomic Rnas ◽  
Transcription System ◽  
Indirect Immunofluorescence Staining

A system has been established to produce infectious RNA transcripts for Striped jack nervous necrosis virus (SJNNV), the type species of the betanodaviruses, which infect fish. An enzymological analysis suggested that both RNA1 and RNA2 of SJNNV have a 5′ cap. Both RNAs were largely resistant to 3′ polyadenylation and ligation, suggesting the presence of an interfering 3′ structure, while a small quantity of viral RNAs were polyadenylated in vitro. The complete 5′ and 3′ non-coding sequences of both segments were determined using the rapid amplification of cDNA ends method. Based on the terminal sequences obtained, RT–PCR was carried out and plasmid clones containing full-length cDNA copies of both RNAs, positioned downstream of a T7 promoter, were constructed. These plasmids were cleaved at a unique restriction site just downstream of the 3′ terminus of each SJNNV sequence and were transcribed in vitro into RNA with a cap structure analogue. A mixture of the transcripts was transfected into the fish cell line E-11. Using indirect immunofluorescence staining with anti-SJNNV serum, fluorescence was observed specifically in these transfected cells; this culture supernatant exhibited pathogenicity to striped jack larvae. Northern blot analysis of E-11 cells infected with the recombinant virus or SJNNV showed small RNA (ca. 0·4 kb) that was newly synthesized and corresponded to the 3′-terminal region of RNA1. Finally, the complete nucleotide sequences of these functional cDNAs (RNA1, 3107 nt; RNA2, 1421 nt) were determined. This is the first report of betanodavirus cDNA clones from which infectious genomic RNAs can be transcribed.

scholarly journals Complete Genome Sequence of Nervous Necrosis Virus Isolated from Orange-Spotted Grouper (Epinephelus coioides) in Taiwan

2017 ◽  
Vol 5 (37) ◽  
Author(s):  
Ming-Hui Chien ◽  
Thi Tuyet Minh Vo ◽  
Shaw-Yun Wu ◽  
Cheng-Hui Lin
Keyword(s):  
Phylogenetic Analysis ◽  
Genome Sequence ◽  
Virus Strain ◽  
Viral Genome ◽  
Nervous Necrosis Virus ◽  
Epinephelus Coioides ◽  
Necrosis Virus ◽  
Virus Genotype ◽  
Rna Molecules ◽  
Positive Sense

ABSTRACT The genome sequence of nervous necrosis virus strain KS1 isolated from orange-spotted grouper (Epinephelus coioides) was cloned and analyzed. The viral genome is composed of two single-stranded positive-sense RNA molecules, RNA1 and RNA2. Phylogenetic analysis shows that the virus strain KS1 belongs to the red-spotted grouper nervous necrosis virus genotype.

scholarly journals Temperature effect on nervous necrosis virus infection in grouper cell line and in grouper larvae

1999 ◽  
Vol 63 (1-2) ◽  
pp. 107-114 ◽  
Author(s):  
Shau-Chi Chi ◽  
Su-Ching Lin ◽  
Huei-Meei Su ◽  
Wei-Wen Hu
Keyword(s):  
Virus Infection ◽  
Temperature Effect ◽  
Cell Line ◽  
Nervous Necrosis Virus ◽  
Necrosis Virus
Sign in / Sign up

Export Citation Format

Share Document