Bunkop Viriyapongsutee, Dokrak Chaisarn, Suphada Kiriratnikom, Nukool Intrasungkha, Poonsook Prasertsan, and Kitjakarn Supamattaya

pp. 275 - 282

Abstract

Fluorescence in situ hybridization technique is very useful for the evaluation of microbial communities in various environments. It is possible to apply this technique to study the intestinal microflora in white
shrimp (Penaeus vannamei). Different fixatives and storage temperature were tested in this technique. It was found that fixation with 10% buffered formalin for 12 hours and changed to 70% ethanol shown positive
results when compared to the fixation with Davidson's fixative or RF fixative. The best signaling was obtained
from the samples which were stored in -20ºC. By using the DNA probe targeted to the Eubacteria domain
(EUB338 probe, 5′-GCT GCC TCC CGT AGG AGT-3′) labeled with fluorescein as a hybridizing probe, it
was found that most intestinal microflora were aggregated with the intestinal contents, or dispersed in the
lumen. There was not evidence of the attachment of the microflora with the intestinal epithelium in this study.