Steps of the Replication Cycle of the Viral Haemorrhagic Septicaemia Virus (VHSV) Affecting Its Virulence on Fish

López-Vázquez, Carmen; Bandı́n, Isabel; Panzarin, Valentina; Toffan, Anna; Cuenca, Argelia; Olesen, Niels Jørgen; Dopazo, Carlos P.

doi:10.3390/ani10122264

Cited by 6 publications

(6 citation statements)

References 43 publications

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“…A cell line is considered susceptible to a given virus when receptors that allow viral attachment are displayed on the cell surface. Therefore, the assessment of the adsorption efficacy is a good measure of cell susceptibility [26,27]. Our AE results indicate that E-11 and DLB-1 (with average values of 90.99 and 94.63%, respectively) are highly susceptible to RGNNV, SJNNV, and RG/SJ strains.…”

Section: Discussionmentioning

confidence: 81%

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

et al. 2021

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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.

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Section: Discussionmentioning

confidence: 81%

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

et al. 2021

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“…From now on, we will refer to these viruses as VHSV-H and VHSV-L, respectively, for VHSV/O.mykiss/I/TN/80/Mar10 and VHSV/O.mykiss/I/TN/480/Oct96. These two viruses, together with several others, were also characterized for virulence determinant and intracellular replication by reverse genetics and in vitro studies, respectively [ 21 , 29 ].…”

Section: Methodsmentioning

confidence: 99%

“…Consequently, from this latter study we have selected two VHSV isolates, with distinct virulence classifications, in order to perform the herein presented experiment. Furthermore, in vitro studies suggest that virulence can mainly be determined by the capacity of the virus to penetrate the host and spread to target organs, and is independent from replication fitness during the infectious process [ 21 ].…”

Section: Introductionmentioning

confidence: 99%

Transcriptome Profiling of Oncorhynchus mykiss Infected with Low or Highly Pathogenic Viral Hemorrhagic Septicemia Virus (VHSV)

Biasini,

Zamperin,

Pascoli

et al. 2023

Microorganisms

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The rainbow trout (Oncorhynchus mykiss) is the most important produced species in freshwater within the European Union, usually reared in intensive farming systems. This species is highly susceptible to viral hemorrhagic septicemia (VHS), a severe systemic disease widespread globally throughout the world. Viral hemorrhagic septicemia virus (VHSV) is the etiological agent and, recently, three classes of VHSV virulence (high, moderate, and low) have been proposed based on the mortality rates, which are strictly dependent on the viral strain. The molecular mechanisms that regulate VHSV virulence and the stimulated gene responses in the host during infection are not completely unveiled. While some preliminary transcriptomic studies have been reported in other fish species, to date there are no publications on rainbow trout. Herein, we report the first time-course RNA sequencing analysis on rainbow trout juveniles experimentally infected with high and low VHSV pathogenic Italian strains. Transcriptome analysis was performed on head kidney samples collected at different time points (1, 2, and 5 days post infection). A large set of notable genes were found to be differentially expressed (DEGs) in all the challenged groups (e.s. trim63a, acod1, cox-2, skia, hipk1, cx35.4, ins, mtnr1a, tlr3, tlr7, mda5, lgp2). Moreover, the number of DEGs progressively increased especially during time with a greater amount found in the group infected with the high VHSV virulent strain. The gene ontology (GO) enrichment analysis highlighted that functions related to inflammation were modulated in rainbow trout during the first days of VHSV infection, regardless of the pathogenicity of the strain. While some functions showed slight differences in enrichments between the two infected groups, others appeared more exclusively modulated in the group challenged with the highly pathogenic strain.

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“…The viral titrations were performed using the endpoint dilution method, as previously described [ 29 ], using 96-well plates with EPC cells (3 replicas per dilution), and the titers determined by the Reed and Müench [ 30 ] procedure, and given as TCID 50 / mL .…”

Section: Methodsmentioning

confidence: 99%

Design and Evaluation of a Macroarray for Detection, Identification, and Typing of Viral Hemorrhagic Septicemia Virus (VHSV)

López-Vázquez

Bandı́n

Dopazo

2021

Animals

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The viral hemorrhagic septicemia virus (VHSV) is the causative agent of an important disease in freshwater and marine fishes. Its diagnosis officially relies on the isolation of the virus in cell culture and its identification by serological or polymerase chain reaction (PCR) methodologies. Nowadays, reverse transcription real-time quantitative PCR (RT-qPCR) is the most widely employed technique for the detection of this virus and some studies have reported the validation of RT-qPCR procedures for the detection, typing, and quantification of VHSV isolates. However, although the efficacy of this technique is not in doubt, it can be cumbersome and even impractical when it comes to processing large numbers of samples, a situation in which cross-contamination problems cannot be ruled out. In the present study, we have designed and validated a macroarray for the simultaneous detection, typing, and quantification of VHSV strains. Its analytical sensitivity (5–50 TCID50/mL), analytical specificity (intra and intergroup), efficiency (E = 100.0–101.1) and reliability (repeatability and reproducibility with CV < 5%, and standard curves with R2 < 0.95) with strains from any VHSV genotype have been widely demonstrated. The procedure is based on the ‘binary multiplex RT-qPCR system (bmRT-qPCR)’ previously reported by the same team, applied to arrays of 96-well PCR strip tubes plates, which can be stored at −25 °C for three months and up to one year before their use, without significant loss of efficiency.

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Steps of the Replication Cycle of the Viral Haemorrhagic Septicaemia Virus (VHSV) Affecting Its Virulence on Fish

Cited by 6 publications

References 43 publications

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

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

Transcriptome Profiling of Oncorhynchus mykiss Infected with Low or Highly Pathogenic Viral Hemorrhagic Septicemia Virus (VHSV)

Design and Evaluation of a Macroarray for Detection, Identification, and Typing of Viral Hemorrhagic Septicemia Virus (VHSV)

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