TABLE 3.

Detection of V. hollisae 525-82Sm^R spiked into APW containing seafood by V. hollisae toxR-specific PCR and by incubation with isolation medium

Seafood sample	Time relative to 4-h incubation	Lowest initial level of 525-82SmR (CFU/ml) detectable by PCRa		Concn (CFU/ml) of purple colonies in APW-seafood homogenateb
				Not spiked		Spiked
		24 cycles	35 cycles	Vh-M	Vh-M + SM	Vh-M	Vh-M + SM
Oysterc	Before	106	105	0	0	2.72 × 103	2.67 × 103
	After	103	102 (5.30)	0	0	7.17 × 104	3.80 × 105
Coastal fish 1d	Before	>106	104	1.80 × 102	0	1.43 × 103	1.85 × 103
	After	104	102 (0.556)	3.00 × 103	0	3.20 × 105	1.84 × 105
Coastal fish 2e	Before	106	103	1.07 × 102	0	5.93 × 102	6.07 × 102
	After	104	10 (2.54)	0	0	9.33 × 103	2.37 × 104
Squidf	Before	>106	105	3.50 × 102	0	1.01 × 104	3.79 × 103
	After	104	102 (1.27)	3.00 × 104	0	1.46 × 105	1.86 × 105
Ivory shellg	Before	105	103	9.33 × 102	0	3.00 × 103	3.73 × 102
	After	104	102 (0.250)	1.03 × 104	0	7.33 × 103	1.87 × 104
Octopusf	Before	>106	105	4.67 × 10	0	4.60 × 102	9.20 × 102
	After	105	103 (1.00)	3.33 × 102	0	8.67 × 103	8.67 × 103
Shrimph	Before	>106	106	5.47 × 102	2.20 × 102	7.87 × 102	2.23 × 102
	After	105	104 (1.40)	7.07 × 104	5.67 × 102	6.5 × 104	9.33 × 103

^aValues in parentheses are ratios of the number of purple colonies on Vh-M supplemented with streptomycin to the number of those on Vh-M recovered from the indicated enrichment culture. 

^bPurple colonies detected on isolation medium. Vh-M, the agar medium designed for isolation of V. hollisae (19). Vh-M + SM, Vh-M supplemented with SM (100 μg/ml). Homogenate was not spiked (not inoculated) or was spiked with 525-82Sm^R (10³ CFU/ml). 

^cCrassostrea gigas. 

^dTrichiurus lepturus. 

^eSillago japonica. 

^fNot identified to the species level. 

^gBabylonia japonica. 

^hPenaeus monodon.