Virulence and Resistance Determinants of Uropathogenic Escherichia coli Strains Isolated from Pregnant and Non-Pregnant Women from Two States in Mexico

Figures & data

Table 1 Antibiotic Resistance of E. coli Strains Isolates from Urine of Pregnant and Non-Pregnant Women from Sonora and Puebla, Mexico

Antibiotics^a	Sonora (n=100) p^b = 0.840					Puebla (n=50) p^b = 0.946
	Pregnant n=50 n (%)	Average %R	Non-Pregnant n=50 n (%)	Average %R	p^c value	Pregnant n=25 n (%)	Average %R	Non-Pregnant n=25 n (%)	Average %R	p^c value
AMK	31 (62)	38	32 (64)	37.3	1	24 (96)	58.7	19 (76)	45.3	0.098
GM	21 (42)		22 (44)		1	14 (56)		10 (40)		0.396
NET	5 (20)		2 (4)		0.436	4 (24)		5 (20)		1
AMP	50 (100)	72.9	50 (100)	78.9	–	100 (25)	82.2	25 (100)	77.3	–
CF	48 (96)		49 (98)		1	24 (96)		25 (100)		1
CFX	41 (82)		50 (100)		0.002	25 (100)		23 (92)		0.489
CTX	35 (70)		47 (94)		0.003	20 (80)		20 (80)		1
CFZ	37 (74)		33 (66)		0.513	21 (84)		19 (76)		0.725
CRO	41 (82)		50 (100)		0.002	24 (96)		24 (96)		1
FEP	19 (38)		14 (28)		0.395	8 (32)		6 (24)		0.753
ATM	20 (40)		15 (30)		0.401	15 (60)		11 (44)		0.396
AMC	37 (74)		47 (94)		0.012	23 (92)		22 (84)		0.667
NA	37 (74)	60	37 (74)	44.8	1	17 (68)	44	18 (72)	56	1
CIP	29 (58)		22 (44)		0.229	10 (40)		13 (52)		0.570
OFX	27 (54)		18 (36)		0.107	12 (48)		14 (56)		0.777
NOR	28 (56)		19 (38)		0.108	13 (32)		14 (56)		0.153
LVX	29 (58)		16 (32)		0.015	13 (32)		11 (44)		0.560
NF	20 (40)	40	41 (82)	82	<0.001	16 (64)	64	17 (68)	68	1
TSX	32 (64)	64	35 (70)	70	0.670	11 (44)	44	17 (68)	68	0.153
FOS	2 (4)	4	2 (4)	4	1	5 (20)	20	1 (4)	4	0.189
C	30 (60)	70	25 (50)	50	0.421	9 (36)	36	11 (44)	44	0.773
TE	35 (70)	70	29 (58)	58	0.032	13 (52)	52	18 (72)	72	0.243
ETP	6 (12)	12	4 (8)	8	0.740	5 (20)	20	0 (0)	0	0.050

Notes: p^b, two samples t-test; p^c, Fisher test exact; –, the value of p could not be obtained; n, number of strains; %, percentage; R, resistance; statistically significant values are in bold.

Abbreviations: AMK, amikacine; GM, gentamicin; NET, netilmicine; AMP, ampicillin; CF, cephalotin; CFX, cefuroxime; CTX, cephotaxime; CFZ, cefazolin; CRO, cephtriaxone; FEP, cephepime; ATM, Aztreonam; AMC, clavulanic acid-ampicillin; NA, nalidixic acid; CIP, ciprofloxacin; OFX, ofloxacin; NOR, norfloxacin; LVX, levofloxacine; NF, nitrofurantoin; TSX, trimethoprim/sulfamethoxazole; FOS, fosfomycin; C, chloramphenicol; TE, tetracycline; ETP, Ertapenem.

Table 2 Antibiotic Profile Classification of E. coli Isolates from Urine of Women from Sonora and Puebla, Mexico in Relation to Pregnant and Non-Pregnant Conditions

City	Condition	Classification by Drug-Resistance n=150 (100%)
		NMDR n (%)	MDR n (%)	XDR n (%)	PDR n (%)
		0 (0)	139 (92.7)	10 (6.7)	1 (0.6)
Puebla	Pregnant (n=25)	0	22 (88)	2 (8)	1 (4)
	Non-pregnant (n=25)	0	24 (96)	1 (4)	0
Sonora	Pregnant (n=50)	0	47 (94)	3 (6)	0
	Non-pregnant (n=50)	0	46 (92)	4 (8)	0

Abbreviations: NMDR, Non-multidrug resistant; MDR, Multidrug-resistance; XDR, extensively multidrug-resistance; PDR, pandrug-resistance.

Figure 1 Multiplex-PCR Banding Patterns of Virulence Genes and Individual PCRcnf of E. coli generated by gel electrophoresis. (A) iucD (512 bp), satA (384 bp), fliC (304 bp) and fimH (210 bp). (B) papGII (562 bp), papGIII (421 bp); vatA (330 bp), vatP (226 bp) and iha (150bp). (C) hlyA (1, 280 bp), satP (880 bp) and papA (641 bp). (D) cnf-1 (3, 100 bp). M. Molecular weight market (100 bp Plus DNA Ladder in A, B and C and 1 kb Plus DNA ladder in D) . Lane C+. Positive control (E. coli CFTO73 in A; E. coli CFTO73 plus E. coli O59I in B and E. coli GAG1 in C and D) . Lines with a number E. coli isolated from the samples. C-. Negative control.

Figure 2 Frequency of the Virulence Genes Among 150 E. coli Isolates From Urine of Pregnant and Non-Pregnant Women From Sonora and Puebla, Mexico. The studied genes encoded the following virulence factor: fimH, type 1 pilus adhesin; papG+papA, adhesin and pilin of the type P pili; iha, enterobactin receptor/Irg homologue adhesin; iucD, aerobactin; satA+satP, autotransporter and peptidase regions of the secreted autotransporter toxin; vatA+vatP, autotransporter and peptidase regions of the vacuolating autotransporter toxin; hlyA, α-hemolysin; cnf-1, cytotoxic necrotizing factor; fliC, flagellin. The statistically significance results (p < 0.05) are in asterisk.

Table 3 Distribution and Percentage of Positivity of PAI in E. coli Strains Isolated from Urine of Pregnant and Non-Pregnant Women from Sonora (A) and Puebla (B), Mexico and Between States and Conditions

PAI	A) Sonora n=51 (51%)
	Pregnant from Sonora n=26	%	Non-Pregnant from Sonora n=25	%	p^b
PAI ICFT073	19	73.1	16	64	0.77
PAI IICFT073	17	65.4	19	76	0.54
PAI IJ96	3	11.5	6	24	0.29
PAI IIJ96	3	11.5	4	16	0.7
PAI	B) Puebla n=34 (68%)			%	pb
	Pregnant from Puebla n=15	%	Non-pregnant from Puebla n=19
PAI ICFT073	12	80	17	89.5	0.63
PAI IICFT073	9	60	9	47.4	0.5
PAI IJ96	5	33.3	4	21	0.46
PAI IIJ96	8	53.3	5	26.3	0.15
PAI	Pregnant from Sonora vs Pregnant from Puebla			%	pb
	Pregnant from Sonora n=26	%	Pregnant from Puebla n=15
PAI ICFT073	19	0.77	12	80	0.71
PAI IICFT073	17	65.4	9	60	0.74
PAI IJ96	3	11.5	5	33.3	0.11
PAI IIJ96	3	11.5	8	53.3	0.008
PAI	Non-Pregnant from Sonora vs Non-Pregnant from Puebla			%	p^b
	Non-Pregnant from Sonora n=25	%	Non-Pregnant from Puebla n=19
PAI ICFT073	16	64	17	89.5	0.081
PAI IICFT073	19	76	9	47.4	0.065
PAI IJ96	6	24	4	21	1
PAI IIJ96	4	16	5	26.3	0.467

Notes: p^b, Fisher test exact; –, the value of p could not be obtained; In bold the statistically significant values.

Abbreviation: PAI, pathogenicity island.

Table 4 E. coli Strains, Isolated from Urine of Pregnant and Non-Pregnant Women from Sonora and Puebla, Associated to Pathotypes or Clinical Cases by the Serotype to Which They Belong

Pathotype or Clinical Case^a	P-SON (%) n=50	NP-SON (%) n=50	Accumulated n=100 (%)	P-PUE (%) n=25	NP-PUE (%) n=25	Accumulated n=50 (%)
UPEC	12 (24)	9 (18)	21 (21)	9 (36)	7 (28)	16 (32)
NT	3 (6)	7 (14)	10 (10)	7 (28)	4 (16)	11 (22)
Unreported	5 (10)	9 (18)	14 (14)	2 (8)	6 (24)	8 (16)
ETEC	–	1 (2)	1 (1)	1 (4)	4 (16)	5 (10)
Heteropathogen	2 (4)	1 (2)	3 (3)	2 (8)	1 (4)	3 (6)
STEC	4 (8)	6 (12)	10 (10)	1 (4)	1 (4)	2 (4)
Isolates from diarrhea	3 (6)	2 (4)	5 (5)	2 (8)	-	2 (4)
EAEC/UPEC	3 (6)	2 (4)	5 (5)	-	2 (8)	2 (4)
Isolates from animals	–	3 (6)	3 (3)	1 (4)	–	1 (2)
EAEC	1 (2)	5 (10)	6 (6)	–	–	–
UPEC or STEC	4 (8)	1 (2)	5 (5)	–	–	–
Isolates from Neonatal sepsis (O20:H9)	9 (18)	4 (8)	13 (13)	–	–	–
Isolates from healthy adults	1 (2)	–	1 (1)	–	–	–
Isolates from pyelonephritis	1 (2)	–	1 (1)	–	–	–
Isolates from SIDS	1 (2)	–	1 (1)	–	–	–
Asociado a HUS	1 (2)	–	1 (1)	–	–	–

Notes: ^aPathotype or clinical case by the serotype to which each strain belongs. Heteropathogen, first strain of hybrid E. coli reported.

Abbreviations: UPEC, uropathogenic E. coli; NT, Notypable; STEC, Shiga toxigenic E. coli; EAEC, enteroaggregative E. coli; EPEC, enteropathogenic E. coli; ETEC, Enterotoxigenic E. coli; SIDS, sudden infant death syndrome; HUS, hemolytic uremic syndrome; P-SON, pregnant women from Sonora; NP-SON, Non-pregnant women from Sonora; P-PUE, pregnant women from Puebla; NP-PUE, non-pregnant women from Puebla.

Table 5 Antibiotic Resistance of E. coli Strains Associated to Pathotypes or Clinical Cases by the Serotype to Which They Belong

Pathotype or Clinical Case^a	Mean of Antibiotic Resistance^b
	P-SON	NP-SON	P-PUE	NP-P
UPEC	15	15	13	15
NT	13	12	15	16
Unreported	14	14	15	16
ETEC	–	10	19	10
Heteropathogen	12	14	15	13
STEC	–	13	12	11
Isolates from diarrhea	16	12	18	–
EAEC, EPEC	11	12	–	16
UPEC, STEC	12	–	–	–
Isolates from animals	–	14	17	–
EAEC	11	14	–	–
Isolates from neonatal sepsis (O20:H9)	20	19	–	-

Notes: ^aPathotype or clinical case by the serotype to which each strain belongs. Heteropathogen, first strain of hybrid E. coli reported; the higher mean of antibiotic resistance are in bold. ^b24 groups of antibiotics tested.

Abbreviations: STEC, shiga toxin productor E. coli; UPEC, uropathogenic E. coli; NT, Notypable; ETEC, Enterotoxigenic E. coli; EAEC, Enteroaggregative E. coli; EPEC, Enteropathogenic E. coli; b, 24 groups of antibiotics tested; P-SON, pregnant women from Sonora; NP-SON, Non-pregnant women from Sonora; P-PUE, pregnant women from Puebla; NP-PUE, Non-pregnant women from Puebla.

Table 6 Virulence Genes of E. coli Strains Associated to Pathotypes or Specific Clinic Cases by the Serotype to Which They Belong

Pathotype or Clinic Case^a	Mean of Virulence Genes^b
	P-SON	NP-SON	P-PUE	NP-PUE
UPEC	5	5	6	3
NT	4	3	5	4
Unreported	4	4	4	5
ETEC	–	4	4	6
Heteropathogen (O2:H6)	5	7	4	7
STEC	2	4	6	2
Isolates from diarrhea	5	4	4	–
Indefinite (EAEC, EPEC, UPEC) (O15:H18)	4	3	–	4
UPEC, STEC	5	–	–	–
EAEC	4	3	–	–
Isolates from neonatal sepsis (O20:H9)	3	4	–	–
Others	5	5	6	–

Notes: ^aPathotype or clinical case by the serotype to which each strain belongs. ^bNine virulence factors tested by PCR; Others, Included Isolates from animals, isolates from healthy adults, isolates from hemolytic uremic syndrome and isolates from sudden infant death syndrome.

Abbreviations: UPEC, Uropathogenic E. coli; ETEC, Enterotoxigenic E. coli; STEC, Shiga toxigenic E. coli; EAEC, enteroaggregative E. coli; EPEC, enteropathogenic E. coli; NT, notypable; P-SON, pregnant women from Sonora; NP-SON, Non-pregnant women from Sonora; P-PUE, pregnant women from Puebla; NP-PUE, Non-pregnant women from Puebla.

Figure 3 Distribution of phylogenetic group among 150 E. coli strains isolated from urine of women from Sonora and Puebla, Mexico. The statistically significance results (p<0.05) are in asterisk.

Abbreviation: CI, Clade I.