Evaluation of the preservation of shrimp samples with Davidson's AFA fixative for infectious myonecrosis virus (IMNV) in situ hybridization

Thales P D Andrade, Rita M. Redman, Donald V Lightner

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

The potential negative effect of prolonged storage of shrimp tissues in Davidson's AFA fixative on in situ hybridization (ISH) signal was demonstrated previously for Taura syndrome virus (TSV), which has a single-stranded RNA genome. In this study we evaluated if prolonged storage of infectious myonecrosis virus (IMNV) infected shrimp in Davidson's AFA (Alcohol, Formaldehyde, Acetic acid) fixative will degrade its double-stranded RNA genome resulting in false negative ISH reactions. Twenty-one shrimp (3 g) specific-pathogen-free Litopenaeus vannamei were used in this study. Three shrimp were used as negative control and 18 shrimp were inoculated with a tissue homogenate prepared from frozen IMNV-infected L. vannamei obtained from Brazil in 2003 (positive control). Shrimp were collected at Day 12 post-injection and fixed in Davidson's AFA for five different preservation times (1, 2, 4, 7 and 10 days). After the different fixation times, the Davidson's AFA was replaced with several changes of 70% ethanol until the pH was stable. IMNV lesions were confirmed in all positive control shrimp by routine H & E histology and ISH. Myonecrosis lesions were strongly positive by ISH at all five preservation times evaluated. Hence, in the present report it was found that the length of time (up to 10 days) in Davidson's AFA did not have a deleterious effect on the ISH reaction for IMNV.

Original languageEnglish (US)
Pages (from-to)179-183
Number of pages5
JournalAquaculture
Volume278
Issue number1-4
DOIs
StatePublished - Jun 10 2008

Fingerprint

formaldehyde
acetic acid
in situ hybridization
alcohol
virus
shrimp
alcohols
viruses
lesion
RNA
sampling
Litopenaeus vannamei
genome
histology
Taura syndrome virus
fixation
ethanol
double-stranded RNA
pathogen
evaluation

Keywords

  • IMNV
  • In situ hybridization
  • Infectious myonecrosis virus
  • Litopenaeus vannamei

ASJC Scopus subject areas

  • Aquatic Science

Cite this

Evaluation of the preservation of shrimp samples with Davidson's AFA fixative for infectious myonecrosis virus (IMNV) in situ hybridization. / Andrade, Thales P D; Redman, Rita M.; Lightner, Donald V.

In: Aquaculture, Vol. 278, No. 1-4, 10.06.2008, p. 179-183.

Research output: Contribution to journalArticle

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