Table 1.

Isolation data, lineage and identification of the new isolates in comparison with those of the control strains.

	Isolation data			Identificationd
Straina	Sourceb	Geographic location/year of isolation	Phylogenetic Lineagec	Species PCR/Api20E	Pathovar/clade E	Public Health Hazard
New isolates
VV3, VV4, VV5	Diseased tilapia	Eastern Mediterranean /2016	?	+/+ (99.3%)	-/-	+
TI417	Diseased tilapia	Eastern Mediterranean /2019	?	+/+ (99.3%)	-/-	+
Control strains
YJ016	Human blood	Asia/1993	L1	+/+ (99.3%)	-/-	+
CECT 529T	Human blood	USA/1980	L2	+/+ (99.3%)	-/-	+
CECT 4999	Diseased eel	Europe/1999	L2/clade E	+/¿? (54.4%)	+/+	+
CECT 5198	Diseased eel	Europe/2000	L2/clade A	+/+ (99.3%)	+/-	-
95-8-161	Diseased eel	Europe/1995	L2/clade I	+/+ (99.3%)	+/-	+

^aCECT, Spanish Type Culture Collection; T, type strain.

^bAll the new isolates were recovered as pure cultures from internal organs of moribund tilapia. VV3 was isolated from farm A, VV4 and VV5 from farm B and TI417 from farm C.

^cPhylogenetic lineage determined by Roig et al [11]. L1 includes biotype 1 strains, L2 biotypes 1 and 2 strains and L3 biotype 3 strains. L2-Clade E includes all the zoonotic strains reported to date.

^dIdentification of the isolates was performed at species, pathovar (piscis) and zoonotic clade (clade E) level by PCR targeting vvhA [20], fpcrp (formerly vep07) [21] and seq 61 [21], respectively. Positive (+), negative (-), and doubtful (¿?) identification. The value in parentheses indicates the probability of a good identification according to the API20E profile (5146105, probability 99.3%; 5006005, probability 54.4%).

^eThe public health hazard of the strain was determined by PCR targeting a polymorphism of pilF [22]. Discrimination is based on the amplification of a variable region located within the gene pilF resulting in a 338 bp fragment.