TABLE 3.

Secretory profile of BLI-Pseudomonas undergoing two treatments with EDTA/taurolidine (assessment of four different phenazines by HPLC-MS)^a,b,c

Treatment	1-Hydroxidyphenazine		Phenazine 1-carboxamide		Phenazine-1-carboxilic acid		Pyocianin
	Peak area	Fold change treated vs untreated	Peak area	Fold change treated vs untreated	Peak area	Fold change treated vs untreated	Peak area	Fold change treated vs untreated
24-h-old biofilm (0–24 h)
TSB	1.26E-06		1.00E-06		8.58E-08		1.24E-09
1st treatment (48–72 h)
SB	1.39E-06	1	2.49E-05	1	9.23E-08	1	1.84E-09	1
EDTA 2.5%	ND		ND		4.13E-07	0.045	8.07E-07	0.044
Tauro 0.5%	ND		ND		2.47E-07	0.027	1.10E-07	0.006
EDTA 2.5% + Tauro 0.5%	ND		ND		2.28E-07	0.025	4.67E-07	0.025
Regrowth (72–96 h)
SB	2.36E-06	1	2.34E-06	1	8.70E-08	1	2.18E-09	1
EDTA 2.5%	2.36E-06	1	1.95E-06	0,83	9.50E-08	1.1	1.73E-09	0.79
Tauro 0.5%	2.25E-06	0.95	1.67E-06	0,71	8.32E-08	1	1,46E-09	0.67
EDTA 2.5% + Tauro 0.5%	2.28E-06	0.97	ND		9.12E-08	1	1.78E-09	0.82
2nd treatment (96–120 h)
SB	2.57E-06	1	1.69E-06	1	1.54E-09	1	1.53E-09	1
EDTA 2.5%	1.08E- 06	0.42	1.18E-06	0,70	6.79E-08	0.44	6.57E-08	0.43
Tauro 0.5%	ND		3.22E-05	0,19	2.64E-07	0.02	ND
EDTA 2.5% + Tauro 0.5%	ND		5.30E-05	0,31	2.85E-07	0.02	ND

^aSupernatants from PDC pieces, contaminated and treated or not treated with EDTA and/or taurolidine as detailed above, were tested by HPLC-MS.

^bThe indicated phenazines were investigated. Using chromatogram analysis, the different elution peaks were identified, and their areas used for semiquantitative evaluation of each specific product. The fold change was calculated with respect to the corresponding SB control. The results shown are from a pool of three replicates of a representative experiment out of two performed.

^cND, not detectable.