Rapid and Sensitive Detection of Lymphocystis Disease Virus Genotype VII by Loop-Mediated Isothermal Amplification

Valverde, Estefanía J.; Cano, Irene; Castro, Dolores; Paley, Richard; Borrego, Juan J.

doi:10.1007/s12560-016-9265-1

Cited by 8 publications

(7 citation statements)

References 31 publications

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“…Viral DNA and transcripts were detected in caudal fin, HK and intestine from 1 dpi, indicating that the virus spreads quickly from the injection site to the skin and internal organs, establishing a systemic infection as it has also been demonstrated in larvae feeding in LCDV‐contaminated rotifers (Cano et al ). The viral load in caudal fin remained low through the experiment, with values similar to those reported in fish sampled from populations suffering subclinical infections (Valverde et al , , ). In HK and intestine, viral loads were significantly lower at the end of the experimental period, which could indicate a decrease in viral replication.…”

Section: Discussionsupporting

confidence: 80%

“…To date, three species have been recognized in the genus, Lymphocystis disease virus 1 (LCDV1) isolated from European flounder ( Platichthys flesus ), Lymphocystis disease virus 2 (LCDV‐C) isolated from Japanese flounder ( Paralichthys olivaceus ) and Lymphocystis disease virus 3 (LCDV‐Sa) isolated from gilthead seabream ( Sparus aurata ) (Chinchar et al ). LCDV‐Sa has also been detected in Senegalese sole ( Solea senegalensis ), both diseased and asymptomatic (Cano et al ; Valverde et al ).…”

Section: Introductionmentioning

confidence: 99%

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Immune gene expression in gilthead seabream ( Sparus aurata ) after Lymphocystis disease virus (LCDV‐Sa) challenge resulting in asymptomatic infection

et al. 2019

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Aims:To determine the immune gene expression response of gilthead seabream (Sparus aurata) that is experimentally infected with the lymphocystivirus LCDV-Sa. Methods and Results: Viral DNA and transcripts were detected by qPCR in all samples from fish injected with LCDV-Sa, demonstrating that the virus establish a systemic and asymptomatic infection. The expression of 23 immune-related genes was also analysed by RT-qPCR in the head kidney (HK) and intestine at several times post-infection (dpi). In HK, the expression of five type I interferon (IFN)-related genes (ifn, irf3, mx2, mx3 and isg15), il10 and ck10 was upregulated at 1-3 dpi, while genes related to the inflammation process (tnfa, il1ß, il6, casp1) were not differentially expressed or even downregulated. The expression profile in the intestine was different regarding type I INF-related genes. An upregulated c3 and ighm expression was observed in both HK and intestine at 3-8 dpi. Finally, the transcription of nccrp1 and mhcIIa was induced in HK, whereas tcrb expression was downregulated in both organs. Conclusions: LCDV-Sa seems to trigger an immune response in gilthead seabream characterized by a partial activation of type I IFN system and a lack of systemic inflammatory response which may be related to viral persistence. Significance and Impact of the Study: The immune response observed in gilthead seabream infected by LCDV-Sa could be implicated in the establishment of an asymptomatic persistent infection.

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Section: Discussionsupporting

confidence: 80%

Section: Introductionmentioning

confidence: 99%

Immune gene expression in gilthead seabream ( Sparus aurata ) after Lymphocystis disease virus (LCDV‐Sa) challenge resulting in asymptomatic infection

et al. 2019

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“…This is the first description of a fish virus that also infects invertebrates. Viral loads and infectious titres estimated in LCDV-infected Artemia were higher than those previously obtained for subclinically infected gilthead seabream fingerlings or juveniles [25,28,38].…”

Section: Discussioncontrasting

confidence: 59%

Artemia spp., a Susceptible Host and Vector for Lymphocystis Disease Virus

Valverde

Labella

Borrego

et al. 2019

Viruses

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Different developmental stages of Artemia spp. (metanauplii, juveniles and adults) were bath-challenged with two isolates of the Lymphocystis disease virus (LCDV), namely, LCDV SA25 (belonging to the species Lymphocystis disease virus 3) and ATCC VR-342 (an unclassified member of the genus Lymphocystivirus). Viral quantification and gene expression were analyzed by qPCR at different times post-inoculation (pi). In addition, infectious titres were determined at 8 dpi by integrated cell culture (ICC)-RT-PCR, an assay that detects viral mRNA in inoculated cell cultures. In LCDV-challenged Artemia, the viral load increased by 2–3 orders of magnitude (depending on developmental stage and viral isolate) during the first 8–12 dpi, with viral titres up to 2.3 × 102 Most Probable Number of Infectious Units (MPNIU)/mg. Viral transcripts were detected in the infected Artemia, relative expression values showed a similar temporal evolution in the different experimental groups. Moreover, gilthead seabream (Sparus aurata) fingerlings were challenged by feeding on LCDV-infected metanauplii. Although no Lymphocystis symptoms were observed in the fish, the number of viral DNA copies was significantly higher at the end of the experimental trial and major capsid protein (mcp) gene expression was consistently detected. The results obtained support that LCDV infects Artemia spp., establishing an asymptomatic productive infection at least under the experimental conditions tested, and that the infected metanauplii are a vector for LCDV transmission to gilthead seabream.

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“…Several approaches have been established to control LCDV outbreaks in the hatcheries in the absence of efficient commercial vaccines. The most widespread approach is the monitoring and removal of asymptomatic carriers by using highly sensitive diagnostic methods [7,8]. This strategy has been successfully applied to prevent vertical transmission, but it requires a continuous evaluation of broodstocks and does not prevent horizontal transmission.…”

Section: Introductionmentioning

confidence: 99%

Heritability Estimates and Genetic Correlation for Growth Traits and LCDV Susceptibility in Gilthead Sea Bream (Sparus aurata)

Carballo

Shin

Berbel

et al. 2019

Fishes

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The lymphocystis disease (LCD) is a viral infection with a high economic impact in gilthead sea bream aquaculture. In this study, genetic estimates associated with lymphocystis disease virus (LCDV) disease susceptibility and growth were determined in sea bream juveniles. Two fish batches (named batch 1 and batch 2) were built from mass spawning and reared under industrial conditions until disease outbreak. At the moment of the sampling (n = 500 specimens for each batch), all animals had the typical LCDV lesions in the skin. For phenotyping, animals were weighted and photographed for image analysis (surface covered and lesion intensity). LCDV DNA copies were quantified in the liver by qPCR. Batch 1 had a higher surface covered and lesion intensity than batch 2, and the body caudal region was the lowest affected region in both batches. The average LCDV DNA copies in liver were higher in the batch 1 than batch 2, and they were positively correlated with severity index (SI) categories (r2 = 0.90–0.94). The total number of families evaluated were 150 and 128 for batch 1 and batch 2, respectively, with a high bias in offspring contribution by family and broodstock. Heritabilities for weight and length were 0.18 and 0.14 in batch 1 and 0.06 and 0.05 in batch 2, respectively. Heritability for the number of viral DNA copies was low (<0.08) in both batches. Heritabilities for SI in binary scale were 0.32/0.33 and 0.21/0.24 (underlying liability/Bayesian approach) for batch 1 and batch 2, respectively. Genetic correlations were very high and positive when growth traits (weight and length) or disease traits (LCDV DNA copies and SI) were compared. In contrast, the genetic correlations between growth and disease traits were moderate–high and positive in the batch 1 but negative in batch 2. These results indicate the genetic selection for LCDV susceptibility and growth is feasible in sea bream juveniles, although estimates are highly dependent on the age. The information provided is relevant to designing selective breeding programs in sea bream.

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Rapid and Sensitive Detection of Lymphocystis Disease Virus Genotype VII by Loop-Mediated Isothermal Amplification

Cited by 8 publications

References 31 publications

Immune gene expression in gilthead seabream ( Sparus aurata ) after Lymphocystis disease virus (LCDV‐Sa) challenge resulting in asymptomatic infection

Immune gene expression in gilthead seabream ( Sparus aurata ) after Lymphocystis disease virus (LCDV‐Sa) challenge resulting in asymptomatic infection

Artemia spp., a Susceptible Host and Vector for Lymphocystis Disease Virus

Heritability Estimates and Genetic Correlation for Growth Traits and LCDV Susceptibility in Gilthead Sea Bream (Sparus aurata)

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