Real-time RT-PCR determination of viral copy number in Penaeus vannamei experimentally infected with Taura syndrome virus

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Abstract

This study examined the viral copy number as determined by real-time RT-PCR, in different tissue samples from Penaeus vannamei exposed to Taura syndrome virus (TSV). Two routes of exposure, injection and per os, were investigated. Six different body parts from each shrimp were assessed for viral copy numbers. Eight shrimp were analyzed per treatment. In addition, eight specific pathogen free (SPF) P. vannamei were analyzed and served as a negative control. The tissue samples examined included: whole tail muscle, tail muscle (shell removed), gills, pleopods, head (cephalothorax with the hepatopancreas included) and tail fan. The results from these experiments showed a significant level of difference between the SPF and the injection treatments. As was expected, there was also a significant difference between the negative control and the per os treatment groups. There was no significant difference between the viral copy number contained in different body parts from the injection experiment. These results contrasted to the per os results, in which there was a statistically significant difference between tail/gills, tail/head, tail/tail fan, whole tail/tail fan and pleopods/tail fan. The tail samples had the lower viral copy numbers, as did the whole tail and pleopods when compared to the tail fan. The mean viral copy number per nanogram of total RNA (x̄ cn/ng tRNA) extracted in the injection study ranged from 1.4 × 105 in the gills to 2.3 × 105 in the whole tail. In the per os experiment, the x̄ cn/ng of extracted tRNA ranged from 2.5 × 10 4 in the tail muscle to 4.3 × 105 in the head. When these values are converted to mean viral copy number per gram (x̄ cn/g) of tissue, the values increased in range. In the injection study, the x̄ cn/g of tissue ranged from 1.2 × 109 in the tail fan to 7.4 × 109 in the head. These values contrast to the x̄ cn/g of tissue in the per os study in that the lowest value of 1.7 × 108 was in the tail muscle and the highest x̄, 1.7 × 1010, was in the head. Overall, all body parts from infected shrimp, regardless of treatment type, quantitatively analyzed by real-time RT-PCR, determined the presence of TSV.

Original languageEnglish (US)
Pages (from-to)1-10
Number of pages10
JournalAquaculture
Volume229
Issue number1-4
DOIs
StatePublished - Jan 12 2004

Fingerprint

Taura syndrome virus
Litopenaeus vannamei
virus
tail
reverse transcriptase polymerase chain reaction
muscle
pathogen
experiment
injection
RNA
muscles
gills
tissue
shrimp
shell
exposure pathways

Keywords

  • Penaeus
  • Real-time PCR
  • Shrimp
  • TSV

ASJC Scopus subject areas

  • Aquatic Science

Cite this

@article{1307a2ebc1844d0e9389a010289a216d,
title = "Real-time RT-PCR determination of viral copy number in Penaeus vannamei experimentally infected with Taura syndrome virus",
abstract = "This study examined the viral copy number as determined by real-time RT-PCR, in different tissue samples from Penaeus vannamei exposed to Taura syndrome virus (TSV). Two routes of exposure, injection and per os, were investigated. Six different body parts from each shrimp were assessed for viral copy numbers. Eight shrimp were analyzed per treatment. In addition, eight specific pathogen free (SPF) P. vannamei were analyzed and served as a negative control. The tissue samples examined included: whole tail muscle, tail muscle (shell removed), gills, pleopods, head (cephalothorax with the hepatopancreas included) and tail fan. The results from these experiments showed a significant level of difference between the SPF and the injection treatments. As was expected, there was also a significant difference between the negative control and the per os treatment groups. There was no significant difference between the viral copy number contained in different body parts from the injection experiment. These results contrasted to the per os results, in which there was a statistically significant difference between tail/gills, tail/head, tail/tail fan, whole tail/tail fan and pleopods/tail fan. The tail samples had the lower viral copy numbers, as did the whole tail and pleopods when compared to the tail fan. The mean viral copy number per nanogram of total RNA (x̄ cn/ng tRNA) extracted in the injection study ranged from 1.4 × 105 in the gills to 2.3 × 105 in the whole tail. In the per os experiment, the x̄ cn/ng of extracted tRNA ranged from 2.5 × 10 4 in the tail muscle to 4.3 × 105 in the head. When these values are converted to mean viral copy number per gram (x̄ cn/g) of tissue, the values increased in range. In the injection study, the x̄ cn/g of tissue ranged from 1.2 × 109 in the tail fan to 7.4 × 109 in the head. These values contrast to the x̄ cn/g of tissue in the per os study in that the lowest value of 1.7 × 108 was in the tail muscle and the highest x̄, 1.7 × 1010, was in the head. Overall, all body parts from infected shrimp, regardless of treatment type, quantitatively analyzed by real-time RT-PCR, determined the presence of TSV.",
keywords = "Penaeus, Real-time PCR, Shrimp, TSV",
author = "Nunan, {L. M.} and Feng-Jyu Tang-Nelson and Lightner, {Donald V}",
year = "2004",
month = "1",
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doi = "10.1016/S0044-8486(03)00365-X",
language = "English (US)",
volume = "229",
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T1 - Real-time RT-PCR determination of viral copy number in Penaeus vannamei experimentally infected with Taura syndrome virus

AU - Nunan, L. M.

AU - Tang-Nelson, Feng-Jyu

AU - Lightner, Donald V

PY - 2004/1/12

Y1 - 2004/1/12

N2 - This study examined the viral copy number as determined by real-time RT-PCR, in different tissue samples from Penaeus vannamei exposed to Taura syndrome virus (TSV). Two routes of exposure, injection and per os, were investigated. Six different body parts from each shrimp were assessed for viral copy numbers. Eight shrimp were analyzed per treatment. In addition, eight specific pathogen free (SPF) P. vannamei were analyzed and served as a negative control. The tissue samples examined included: whole tail muscle, tail muscle (shell removed), gills, pleopods, head (cephalothorax with the hepatopancreas included) and tail fan. The results from these experiments showed a significant level of difference between the SPF and the injection treatments. As was expected, there was also a significant difference between the negative control and the per os treatment groups. There was no significant difference between the viral copy number contained in different body parts from the injection experiment. These results contrasted to the per os results, in which there was a statistically significant difference between tail/gills, tail/head, tail/tail fan, whole tail/tail fan and pleopods/tail fan. The tail samples had the lower viral copy numbers, as did the whole tail and pleopods when compared to the tail fan. The mean viral copy number per nanogram of total RNA (x̄ cn/ng tRNA) extracted in the injection study ranged from 1.4 × 105 in the gills to 2.3 × 105 in the whole tail. In the per os experiment, the x̄ cn/ng of extracted tRNA ranged from 2.5 × 10 4 in the tail muscle to 4.3 × 105 in the head. When these values are converted to mean viral copy number per gram (x̄ cn/g) of tissue, the values increased in range. In the injection study, the x̄ cn/g of tissue ranged from 1.2 × 109 in the tail fan to 7.4 × 109 in the head. These values contrast to the x̄ cn/g of tissue in the per os study in that the lowest value of 1.7 × 108 was in the tail muscle and the highest x̄, 1.7 × 1010, was in the head. Overall, all body parts from infected shrimp, regardless of treatment type, quantitatively analyzed by real-time RT-PCR, determined the presence of TSV.

AB - This study examined the viral copy number as determined by real-time RT-PCR, in different tissue samples from Penaeus vannamei exposed to Taura syndrome virus (TSV). Two routes of exposure, injection and per os, were investigated. Six different body parts from each shrimp were assessed for viral copy numbers. Eight shrimp were analyzed per treatment. In addition, eight specific pathogen free (SPF) P. vannamei were analyzed and served as a negative control. The tissue samples examined included: whole tail muscle, tail muscle (shell removed), gills, pleopods, head (cephalothorax with the hepatopancreas included) and tail fan. The results from these experiments showed a significant level of difference between the SPF and the injection treatments. As was expected, there was also a significant difference between the negative control and the per os treatment groups. There was no significant difference between the viral copy number contained in different body parts from the injection experiment. These results contrasted to the per os results, in which there was a statistically significant difference between tail/gills, tail/head, tail/tail fan, whole tail/tail fan and pleopods/tail fan. The tail samples had the lower viral copy numbers, as did the whole tail and pleopods when compared to the tail fan. The mean viral copy number per nanogram of total RNA (x̄ cn/ng tRNA) extracted in the injection study ranged from 1.4 × 105 in the gills to 2.3 × 105 in the whole tail. In the per os experiment, the x̄ cn/ng of extracted tRNA ranged from 2.5 × 10 4 in the tail muscle to 4.3 × 105 in the head. When these values are converted to mean viral copy number per gram (x̄ cn/g) of tissue, the values increased in range. In the injection study, the x̄ cn/g of tissue ranged from 1.2 × 109 in the tail fan to 7.4 × 109 in the head. These values contrast to the x̄ cn/g of tissue in the per os study in that the lowest value of 1.7 × 108 was in the tail muscle and the highest x̄, 1.7 × 1010, was in the head. Overall, all body parts from infected shrimp, regardless of treatment type, quantitatively analyzed by real-time RT-PCR, determined the presence of TSV.

KW - Penaeus

KW - Real-time PCR

KW - Shrimp

KW - TSV

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