TY - JOUR
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, K.
AU - Lightner, D. V.
N1 - Funding Information:
Funding for this research was provided by a grant from the USMSFP, USDA, CREES, Grant No. 99-38808-7431. The authors would like to thank Brenda White-Noble, Dr. Carlos Pantoja, Leone Mohney, and Rita Redman for their input and/or assistance in conducting these experiments and Doug Jones and Wayne Nicholson for their advice on evaluation of the statistical data.
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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U2 - 10.1016/S0044-8486(03)00365-X
DO - 10.1016/S0044-8486(03)00365-X
M3 - Article
AN - SCOPUS:0347664070
SN - 0044-8486
VL - 229
SP - 1
EP - 10
JO - Aquaculture
JF - Aquaculture
IS - 1-4
ER -