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TaqMan real-time PCR assay for relative quantitation of white spot syndrome virus infection in Penaeus monodon Fabricius exposed to ammonia.

White spot disease is caused by a highly virulent pathogen, the white spot syndrome virus (WSSV). The disease is usually triggered by changes in environmental parameters causing severe losses to the shrimp industry. This study was undertaken to quantify the relative WSSV load in shrimp exposed to am...

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Main Authors: M., Fouzi, Mohamed Din, Mohamed Shariff, Omar, Abdul Rahman, Md Yusoff, Fatimah, Tan, S. W.
Format: Article
Language:English
English
Published: Blackwell Publishing Ltd 2010
Online Access:http://psasir.upm.edu.my/id/eprint/16963/1/TaqMan%20real.pdf
http://psasir.upm.edu.my/id/eprint/16963/
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spelling my.upm.eprints.169632015-10-21T23:43:03Z http://psasir.upm.edu.my/id/eprint/16963/ TaqMan real-time PCR assay for relative quantitation of white spot syndrome virus infection in Penaeus monodon Fabricius exposed to ammonia. M., Fouzi Mohamed Din, Mohamed Shariff Omar, Abdul Rahman Md Yusoff, Fatimah Tan, S. W. White spot disease is caused by a highly virulent pathogen, the white spot syndrome virus (WSSV). The disease is usually triggered by changes in environmental parameters causing severe losses to the shrimp industry. This study was undertaken to quantify the relative WSSV load in shrimp exposed to ammonia, using a TaqMan-based real-time PCR, and their subsequent susceptibility to WSSV. Shrimp were exposed to different levels of total ammonia nitrogen (TAN) (8.1, 3.8 and 1.1 mg L−1) for 10 days and challenged with WSSV by feeding WSSV-positive shrimp. WSSV was detected simultaneously in haemolymph, gills and pereopods at four hours post-infection. The TaqMan real-time PCR assay showed a highly dynamic detection limit that spanned over 6 log10 concentrations of DNA and high reproducibility (standard deviation 0.33–1.42) and small correlation of variability (CV) (1.89–3.85%). Shrimp exposed to ammonia had significantly higher (P < 0.01) WSSV load compared to the positive control, which was not exposed to ammonia. Shrimp exposed to 8.1 mg L−1 of TAN had the highest (P < 0.01) WSSV load in all three organs in comparison with those exposed to 3.8 and 1.1 mg L−1 of TAN. However, haemolymph had significantly higher (P < 0.01) viral load compared to the gills and pereopods. Results showed that shrimp exposed to ammonia levels as low as 1.1 mg L−1 (TAN) had increased susceptibility to WSSV. Blackwell Publishing Ltd 2010 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/16963/1/TaqMan%20real.pdf M., Fouzi and Mohamed Din, Mohamed Shariff and Omar, Abdul Rahman and Md Yusoff, Fatimah and Tan, S. W. (2010) TaqMan real-time PCR assay for relative quantitation of white spot syndrome virus infection in Penaeus monodon Fabricius exposed to ammonia. Journal of Fish Diseases, 33. pp. 931-938. ISSN 0140-7775 10.1111/j.1365-2761.2010.01200.x English
institution Universiti Putra Malaysia
building UPM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Putra Malaysia
content_source UPM Institutional Repository
url_provider http://psasir.upm.edu.my/
language English
English
description White spot disease is caused by a highly virulent pathogen, the white spot syndrome virus (WSSV). The disease is usually triggered by changes in environmental parameters causing severe losses to the shrimp industry. This study was undertaken to quantify the relative WSSV load in shrimp exposed to ammonia, using a TaqMan-based real-time PCR, and their subsequent susceptibility to WSSV. Shrimp were exposed to different levels of total ammonia nitrogen (TAN) (8.1, 3.8 and 1.1 mg L−1) for 10 days and challenged with WSSV by feeding WSSV-positive shrimp. WSSV was detected simultaneously in haemolymph, gills and pereopods at four hours post-infection. The TaqMan real-time PCR assay showed a highly dynamic detection limit that spanned over 6 log10 concentrations of DNA and high reproducibility (standard deviation 0.33–1.42) and small correlation of variability (CV) (1.89–3.85%). Shrimp exposed to ammonia had significantly higher (P < 0.01) WSSV load compared to the positive control, which was not exposed to ammonia. Shrimp exposed to 8.1 mg L−1 of TAN had the highest (P < 0.01) WSSV load in all three organs in comparison with those exposed to 3.8 and 1.1 mg L−1 of TAN. However, haemolymph had significantly higher (P < 0.01) viral load compared to the gills and pereopods. Results showed that shrimp exposed to ammonia levels as low as 1.1 mg L−1 (TAN) had increased susceptibility to WSSV.
format Article
author M., Fouzi
Mohamed Din, Mohamed Shariff
Omar, Abdul Rahman
Md Yusoff, Fatimah
Tan, S. W.
spellingShingle M., Fouzi
Mohamed Din, Mohamed Shariff
Omar, Abdul Rahman
Md Yusoff, Fatimah
Tan, S. W.
TaqMan real-time PCR assay for relative quantitation of white spot syndrome virus infection in Penaeus monodon Fabricius exposed to ammonia.
author_facet M., Fouzi
Mohamed Din, Mohamed Shariff
Omar, Abdul Rahman
Md Yusoff, Fatimah
Tan, S. W.
author_sort M., Fouzi
title TaqMan real-time PCR assay for relative quantitation of white spot syndrome virus infection in Penaeus monodon Fabricius exposed to ammonia.
title_short TaqMan real-time PCR assay for relative quantitation of white spot syndrome virus infection in Penaeus monodon Fabricius exposed to ammonia.
title_full TaqMan real-time PCR assay for relative quantitation of white spot syndrome virus infection in Penaeus monodon Fabricius exposed to ammonia.
title_fullStr TaqMan real-time PCR assay for relative quantitation of white spot syndrome virus infection in Penaeus monodon Fabricius exposed to ammonia.
title_full_unstemmed TaqMan real-time PCR assay for relative quantitation of white spot syndrome virus infection in Penaeus monodon Fabricius exposed to ammonia.
title_sort taqman real-time pcr assay for relative quantitation of white spot syndrome virus infection in penaeus monodon fabricius exposed to ammonia.
publisher Blackwell Publishing Ltd
publishDate 2010
url http://psasir.upm.edu.my/id/eprint/16963/1/TaqMan%20real.pdf
http://psasir.upm.edu.my/id/eprint/16963/
_version_ 1643826371685777408
score 13.211869

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