2.4. Enumeration of V. vulnificus
Initial V. vulnificus loads were determined at day 0 (prior to
depuration but post temperature abuse), and at days 3, 6, 10 and 14
during the depuration process. Twelve oysters were analyzed each
time according to the 3-tube Most Probable Number (MPN) method
described in the U.S. Food and Drug Administration’s Bacteriological
Analytical Manual (BAM) (FDA, 2002). The MPN procedure
was followed by a colony lift and subsequent colony blot hybridization
using a V. vulnificus-specific oligonucleotide probe (vvhA)
(DNA Technology, Aarhus, Denmark) that targets the hemolysin
gene as previously described (Wright et al., 1993). Numbers of
V. vulnificus are given as MPN/g of oyster tissue. For samples
resulting in less than 3 MPN/g, 3 MPN/g was used for statistical
analysis